Study of the Analytical Performance of ALLEGRO™ Instrument: Performance Evaluation of In Vitro Diagnostic Device
Conditions: Diabetes
Sponsor: Nova Biomedical

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Version 3: 18 July 2016 
[Institution]                                          [Place Institution Logo] 
 
 
 
 
 
 
 
 
 
 
Allegro Evaluation Protocols 
 
Nova Biomedical |Medical & Scientific Affairs 
 
 
 
 
 
 
 
Version 3: 18 July 2016

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Preface 
Clinical laboratory based method evaluations are a set of studies aimed at characterizing the performance of an 
assay – a procedure for qualitatively assessing or quantitatively measuring the presence or amount of a target 
entity, i.e. analyte or measurand.   These studies utilize controlled experimental technique to assess the various 
components of a measurement including the assessment of: method comparison, reproducibility, specificity, 
and accuracy. Highly accurate and precise measurements are central to the practice of laboratory medicine 
given that these measurements are used to manage the patient throughout the entire spectrum of care. It is 
therefore necessary that clinical laboratorians perform their due diligence in the careful selection and 
subsequent management of laboratory methods. In order to assist the investigator, a collection of evaluation 
protocols specifically designed to assess the performance of the Allegro has been provided. These protocols are 
based on the recommendations provided by the Clinical and Laboratory Standards Institute (CLSI), a recognized 
body that publishes consensus guidelines on the various practice aspects of laboratory medicine.  
 
 
 
 
 
 
 
 
 

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Version 3: 18 July 2016 
 
Introduction 
Allegro is a fast, simple, capillary whole blood analyser with a diabetes-focused menu. 
The comprehensive test menu available on the Allegro is: 
Glycemic Control 
Lipid  
            Kidney Function 
 
HbA1c  
 
Total Cholesterol 
Urine Creatinine 
 
 
 
 
HDL Cholesterol 
Urine Albumin  
 
Triglyceride 
The Allegro can be used in conjunction with the companion tests below to further extend the diabetes 
management menu  
Companion Tests 
Anemia – Hemoglobin and Hematocrit  
Glycemic control – Glucose and Ketone 
 
HbA1c, Lipid and Kidney Function tests are provided in their own multi test cartridge so each cartridge 
measures all analytes within each profile 
Scope 
The scope of this protocol is to evaluate and assess the analytical performance of the HbA1c, Lipid cartridge 
and the Kidney Function cartridge.  
Analytical performance will be assessed on each profile e.g. Lipid, HbA1c, and Kidney Function, independently 
and will comprise of: 
Within Day precision 
Between Day precision 
Method Correlation 
 
Supplemental System Specifications: 
Operating Environment: 
Operating Location: 
The analyzer is intended for indoor operation on a stationary, stable, bench. 
Operating Temperature and Relative Humidity Range: 
The Analyzer shall be able to operate under the following conditions 
• 
Operating Temperature Range:  
15 to 32°C (59°F to 90°F) 
• 
Relative Humidity Range: 
 
20 – 85%, non-condensing  
Altitude: 
The Analyzer shall be able to operate at an altitude of up to 3,660 meters (12,000 feet)  
 

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Version 3: 18 July 2016 
 
Power Requirements: 
Operating Voltage: 
90 to 264 VAC 
Operating Frequency: 50/60Hz 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Contents 
1. Kidney Function (Tests: Urine Creatinine and Albumin) 
I. 
Within Day Precision Study Protocol  
II. 
Between Day Precision Study Protocol 
III. 
Method Comparison Protocol  
IV. 
Summary Report Template  
 
2. Glycemic Control (Tests: HbA1c) 
I. 
Within Day Precision Study Protocol 
II. 
Between Day Precision Study Protocol 
III. 
Method Comparison Study Protocol  
IV. 
Summary Report Template  
 
3. Lipid (Tests: Total Cholesterol, Triglycerides, HDL Cholesterol 
I. 
Within Day Precision Study Protocol 
II. 
Between Day Precision Study Protocol 
III. 
Method Comparison Study Protocol  
IV. 
Summary Report Template  
 
 
 
 
 
 
 

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1. Urine Creatinine and Albumin 
 
 
 
 
 
I. 
Within Day Precision Study Protocol 
Protocol based on: 
 
Clinical & Laboratory Standards Institute (CLSI). Evaluation of Precision Performance of Quantitative 
Measurement Methods; Approved Guideline – Second Edition. EP5-A2. 
 
 
 
 

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Purpose 
This protocol is recommended and designed by Nova Biomedical to investigate the within day precision of urine 
creatinine and urine Albumin measurements on the Allegro system. 
Materials Needed 
 
 Nova Biomedical Allegro 
 Allegro Kidney Function Cartridges 
 Nova Biomedical Allegro Quality Control (QC) Solution 
 Reference Analyzer and reagents 
 2mL plastic tubes for decanting urine samples 
 Plastic 2mL disposable pipettes 
 Full range of adjustable pipettes and tips 
 If required appropriate Spiking stock solutions 
 
Important Factors to Consider 
Tests: Urine Creatinine and Urine Albumin 
Sample Type: Fresh random urine 
Sample Size: 25µL 
Time per UACR: 7 mins 
Linear Range of Operation: 15-500 mg/dL (13.0-44.2 mmol/L) 
Linear Range of Operation for urine Albumin: 5-300 mg/L 
 
Expected Normal Values 
Urine Creatinine: For a random urine sample, >12 Years Male 20-370 mg/dL (1.8- 32.7 mmol/L), >12 Years 
Female 20 – 320 mg/dL (1.8-28.3 mmol/L) 
Urine Albumin: For a random urine sample, normal values are typically 0 to 30 mg/L 
 
Control Levels: 
The Allegro supports two levels of controls.  The following are suggested ranges for each parameter and level: 
Urine Albumin and Creatinine: 
Low Level Target: 
 
Urine Albumin ~50 mg/L 
 
Creatinine ~85 mg/dL (~8 mmol/L) 
High Level Target: 
 
Urine Albumin ~180 mg/L 
 
Creatinine ~220 mg/dL (~20 mmol/L) 
 
Please note that urine creatinine and Albumin are processed at the same time and can be run in both channels. 
 
 
 
 

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Procedure 
1. Fill out the requested information (Allegro serial numbers, cartridge lot numbers, QC lot numbers, QC 
ranges) on the provided Data Sheets (see Appendix).   
 
2. Perform quality control runs on the Allegro being evaluated and the reference analyzer. Record the 
results for all systems on the Data Sheet and make sure the results are within the manufacturer’s 
control range. Proceed with testing only if all control results are within range.  
 
3. Select low, med and high levels of both urine creatinine and Albumin, ideally using discarded random 
urine collections or, if these are not available, then urine samples that have been spiked with 
appropriate concentrations of creatinine spiking solution and/or human Albumin (may need to be 
tested for homogeneity).     
 
4. Suggested concentrations for low, medium and high samples: - 
 
 
 
 
Urine Creatinine 
 
 
  
Urine Albumin 
Low Target 
 
 
~25-30 mg/dL (~2.2-2.7mmol/L) 
 
~10-15 mg/L 
Medium Target 
 
~200 mg/dL (~18 mmol/L) 
 
 
~40 mg/L 
High Target 
 
 
~ 400-450 mg/dL (~35-40 mmol/L) 
 
~150-200 mg/L 
 
5. Ensure adequate mixing before analysis. 
 
6. Cartridges are stored at 4° each cartridge requires 10-15 mins warm up time so consider this requirement 
when planning study needs. 
 
7. Run each Level 10 times on Allegro, consider running only in one channel to reduce variability. Record results 
on Data Collection Sheet.   
 
8. Total Analysis Time: 7 mins x 10 = 70 mins per level. 210 mins for all levels. Approx. 3.5 hrs plus preparation 
time.  
Data Analysis 
The within-run precision of the Allegro be assessed by calculating the % coefficient of variation (%CV) from the 
mean and standard deviation from the replicate measurements. Acceptability criteria are determined by the 
medical director and/or institution. Investigators are referred to organizations such as CLSI and the 
International Standards Organization (ISO) which offer published acceptance criteria [1]. 
 
Data Review 
This data will be summarized in a final report which will also include an assessment of the accuracy. This report 
should be reviewed by the study site principal investigator. Results of this laboratory evaluation can be used to 
document verification of Allegro performance prior to the conductance of clinical studies. 
 
 
 

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References 
 
1.  Clinical & Laboratory Standards Institute (CLSI). Evaluation of Precision Performance of Quantitative  
             Measurement Methods; Approved Guideline – Second Edition. EP5-A2. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Appendix 1 
 
Within-Day Precision Study Data Collection Sheet                                                                                   
 
Hospital Name: _____________________________ Date of Study: _____________________ 
System Operator(s): ____________________________________________________________ 
Reference Analyzer ______________________ 
 
Level 1 Urine Creatinine and Urine Albumin 
 
Reference Analyzer Result: Urine Creatinine mg/dL or mmol/L __________ 
Reference Analyzer Result: Urine Albumin mg/L or g/L____________ 
 
 
System Name 
 
 
 
 
 
 
 
System Serial # 
 
 
 
 
 
 
 
Cartridge Lot # 
 
 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
 
 
Replicate # 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine Albumin 
Result 
mg/L or g/L 
 
 
 
 
 
1 
 
 
 
 
 
 
 
2 
 
 
 
 
 
 
 
3 
 
 
 
 
 
 
 
4 
 
 
 
 
 
 
 
5 
 
 
 
 
 
 
 
6 
 
 
 
 
 
 
 
7 
 
 
 
 
 
 
 
8 
 
 
 
 
 
 
 
9 
 
 
 
 
 
 
 
10 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Level 2 Urine Creatinine and Urine Albumin 
 
Reference Analyzer Result: Urine Creatinine mg/dL or mmol/L __________ 
Reference Analyzer Result: Urine Albumin mg/L or g/L____________ 
 
 
System Name 
 
 
 
 
 
 
 
System Serial # 
 
 
 
 
 
 
 
Cartridge Lot # 
 
 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
 
 
Replicate # 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine Albumin 
Result 
mg/L or g/L 
 
 
 
 
 
1 
 
 
 
 
 
 
 
2 
 
 
 
 
 
 
 
3 
 
 
 
 
 
 
 
4 
 
 
 
 
 
 
 
5 
 
 
 
 
 
 
 
6 
 
 
 
 
 
 
 
7 
 
 
 
 
 
 
 
8 
 
 
 
 
 
 
 
9 
 
 
 
 
 
 
 
10 
 
 
 
 
 
 
 
 
 
Level 3 Urine Creatinine and Urine Albumin 
 
Reference Analyzer Result: Urine Creatinine mg/dL or mmol/L __________ 
Reference Analyzer Result: Urine Albumin mg/L or g/L____________ 
 
 
System Name 
 
 
 
 
 
 
 
System Serial # 
 
 
 
 
 
 
 
Cart Lot # 
 
 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
 
 
Replicate # 
Urine Creat 
Result mg/dL or 
mmol/L 
Urine Albumin 
Result 
mg/L or g/L 
 
 
 
 
 
1 
 
 
 
 
 
 
 
2 
 
 
 
 
 
 
 
3 
 
 
 
 
 
 
 
4 
 
 
 
 
 
 
 
5 
 
 
 
 
 
 
 
6 
 
 
 
 
 
 
 
7 
 
 
 
 
 
 
 
8 
 
 
 
 
 
 
 
9 
 
 
 
 
 
 
 
10 
 
 
 
 
 
 
 
 
Name of Person Collecting Data: _______________________________ 
Signature of Person Collecting Data: ____________________________ 
Date: ________________________ 

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I. Urine Creatinine and Urine Albumin 
 
II. Between-Day Precision Study Protocol 
 
Protocol based on: 
Clinical & Laboratory Standards Institute (CLSI). Evaluation of Precision Performance of Quantitative 
Measurement Methods; Approved Guideline – Second Edition. EP5-A2. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Version 3: 18 July 2016 
 
Purpose 
This protocol is recommended and designed by Nova Biomedical to investigate the between-day precision of 
urine creatinine and urine Albumin measurements on the Allegro system. 
Materials Needed 
 
 Nova Biomedical Allegro 
 Allegro Kidney Function Cartridges 
 Nova Biomedical Allegro Quality Control (QC) Solution 
 Reference Analyzer and reagents 
 2mL plastic tubes for decanting urine samples 
 Plastic 2mL disposable pipettes 
 Full range of adjustable pipettes and tips 
 If required appropriate Spiking stock solutions 
 
Important Factors to Consider 
Tests: Urine Creatinine and Urine Albumin 
Sample Type: Fresh random urine 
Sample Size: 25µL 
Time per UABCR: 7 mins 
Linear Range of Operation for Urine Creatinine: 15-500 mg/dL (13.0-44.2 mmol/L) 
Linear Range of Operation for Urine Albumin: 3-300 mg/L 
 
Expected Normal Values 
Urine Creatinine: For a random urine sample, >12 Years Male 20-370 mg/dL (1.8- 32.7 mmol/L), >12 Years 
Female 20 – 320 mg/dL (1.8-28.3 mmol/L) 
Urine Albumin: For a random urine sample, normal values are typically 0 to 30 mg/dL 
 
Control Levels: 
The Allegro supports two levels of controls.  The following are suggested ranges for each parameter and level: 
Urine Albumin and Creatinine: 
Low Level Target: 
 
Urine Albumin ~50 mg/L 
 
Creatinine ~85 mg/L (~8 mmol/L) 
High Level Target: 
 
Urine Albumin ~180 mg/L 
 
Creatinine ~220 mg/L (~20 mmol/L) 
 
Please note that urine creatinine and urine Albumin are processed at the same time 
 
 
 
 

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Version 3: 18 July 2016 
 
Procedure 
1. Fill out the requested information (Allegro serial numbers, cartridge lot numbers, QC lot numbers, QC 
ranges) on the provided Data Sheets (see Appendix).   
 
2. Perform quality control runs on the Allegro being evaluated and the reference analyzer. Analyse 
controls on Allegro every day that testing is carried out, 2 runs per day would be performed one in the 
morning and the other in the afternoon so QC would need to be performed prior to each run, make sure 
results are within specification before proceeding. Record the results for all systems on the Data Sheet 
and make sure the results are within the manufacturer’s control range. Proceed with testing only if all 
control results are within range.  
 
3. Select low, med and high levels of both urine creatinine and Albumin. These can be obtained from one 
or more of the following: a) samples from random urine collections, b) samples spiked with stock 
concentrations of Albumin and/or creatinine, or c) samples used for the within day precision test. Forty 
aliquots must be prepared from each sample and frozen.  
 
4. A freshly thawed aliquot will then be analysed every morning and afternoon in duplicate for 10 
consecutive working days 
 
5. Suggested concentrations for low, medium and high samples: - 
 
 
 
 
Urine Creatinine 
 
 
 
Urine Albumin 
Low Target 
 
 
~25-30 mg/dL     (~2.2-2.7 mmol/L) 
 
~10-15 mg/L 
Medium Target 
 
~200 mg/dL 
    (~18 mmol/L) 
 
~40 mg/L 
High Target 
 
 
~ 400-450 mg/dL (~35-40 mmol/L) 
 
~150-200 mg/L 
 
6. Ensure adequate mixing before analysis. 
 
7. Cartridges are stored at 4° each cartridge requires 10-15 mins warm up time so consider this requirement 
when planning study needs. 
 
8. Run each level in duplicate on the Allegro using the same channel to reduce variability 
 
9. Collect, review and collate data 
 
10. Total Analysis Time: 7 mins x 6 = 42 mins in morning, 7 mins x 6 = 42 mins in afternoon plus preparation 
time   
Data Analysis 
The between-day precision of the Allegro be assessed by calculating the % coefficient of variation (%CV) from 
the mean and standard deviation from the replicate measurements. Acceptability criteria are determined by 
the medical director and/or institution. Investigators are referred to organizations such as CLSI and the 
International Standards Organization (ISO) which offer published acceptance criteria [1, 2].  
 

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Version 3: 18 July 2016 
 
 
Data Review 
This data will be summarized in a final report which will also include an assessment of the accuracy. This report 
should be reviewed by the study site principal investigator. Results of this laboratory evaluation can be used to 
document verification of Allegro performance prior to the conductance of clinical studies. 
References 
 
1.  Clinical & Laboratory Standards Institute (CLSI). Evaluation of Precision Performance of Quantitative  
        Measurement Methods; Approved Guideline – Second Edition. EP5-A2. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Appendix 2 

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Version 3: 18 July 2016 
 
 
Between-Day Precision Study Data Collection Sheet                                                                                   
 
Hospital Name: _____________________________ Date of Study: _____________________ 
System Operator(s): ____________________________________________________________ 
Reference Analyzer ______________________ 
 
Level 1 Urine Creatinine and Urine Albumin 
Morning 
Reference Analyzer Result: Urine Creatinine mg/dL or mmol/L __________ 
Reference Analyzer Result: Urine Albumin mg/L or g/L____________ 
Afternoon 
Reference Analyzer Result: Urine Creatinine mg/dL or mmol/L __________ 
Reference Analyzer Result: Urine Albumin mg/L or g/L____________ 
 
 
 
System Name 
 
 
 
 
 
 
 
 
System Serial # 
 
 
 
 
 
 
 
 
Cartridge Lot # 
 
 
 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
 
 
 
 
Morning 
Afternoon 
 
Replicate # 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Day 1 
 
 
 
 
 
 
 
 
Day 2 
 
 
 
 
 
 
 
 
Day 3 
 
 
 
 
 
 
 
 
Day 4 
 
 
 
 
 
 
 
 
Day 5 
 
 
 
 
 
 
 
 
Day 6 
 
 
 
 
 
 
 
 
Day 7 
 
 
 
 
 
 
 
 
Day 8 
 
 
 
 
 
 
 
 
Day 9 
 
 
 
 
 
 
 
 
Day 10 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Level 2 Urine Creatinine and Urine Albumin 
 
Morning 
Reference Analyzer Result: Urine Creatinine mg/dL or mmol/L __________ 
Reference Analyzer Result: Urine Albumin mg/L or g/L____________ 
Afternoon 
Reference Analyzer Result: Urine Creatinine mg/dL or mmol/L __________ 
Reference Analyzer Result: Urine Albumin mg/L or g/L____________ 
 
 
System Name 
 
 
 
 
 
 
 
 
System Serial # 
 
 
 
 
 
 
 
 
Cartridge Lot # 
 
 
 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
 
 
 
 
Morning 
Afternoon 
 
Replicate # 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Day 1 
 
 
 
 
 
 
 
 
Day 2 
 
 
 
 
 
 
 
 
Day 3 
 
 
 
 
 
 
 
 
Day 4 
 
 
 
 
 
 
 
 
Day 5 
 
 
 
 
 
 
 
 
Day 6 
 
 
 
 
 
 
 
 
Day 7 
 
 
 
 
 
 
 
 
Day 8 
 
 
 
 
 
 
 
 
Day 9 
 
 
 
 
 
 
 
 
Day 10 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Level 3 Urine Creatinine and Urine Albumin 

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Morning 
Reference Analyzer Result: Urine Creatinine mg/dL or mmol/L __________ 
Reference Analyzer Result: Urine Albumin mg/L or g/L____________ 
Afternoon 
Reference Analyzer Result: Urine Creatinine mg/dL or mmol/L __________ 
Reference Analyzer Result: Urine Albumin mg/L or g/L____________ 
 
 
System Name 
 
 
 
 
 
 
 
 
System Serial # 
 
 
 
 
 
 
 
 
Cartridge Lot # 
 
 
 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
 
 
 
 
Morning 
Afternoon 
 
Replicate # 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Urine 
Albumin 
Result 
mg/L or gl/L 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine 
Albumin 
Result 
mg/L or gl/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Day 1 
 
 
 
 
 
 
 
 
Day 2 
 
 
 
 
 
 
 
 
Day 3 
 
 
 
 
 
 
 
 
Day 4 
 
 
 
 
 
 
 
 
Day 5 
 
 
 
 
 
 
 
 
Day 6 
 
 
 
 
 
 
 
 
Day 7 
 
 
 
 
 
 
 
 
Day 8 
 
 
 
 
 
 
 
 
Day 9 
 
 
 
 
 
 
 
 
Day 10 
 
 
 
 
 
 
 
 
 
Name of Person Collecting Data: _______________________________ 
Signature of Person Collecting Data: ____________________________ 
Date: ________________________ 
 
 
 
 
 
 
 
 
 
 
 
 

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I. Urine Creatinine and Urine Albumin 
 
 
 
 
 
III. 
Method Comparison Study Protocol 
 
Protocol based on the following guidance documents: 
 
Clinical & Laboratory Standards Institute (CLSI). Method Comparison and Bias Estimation Using Patient Samples; 
Approved Guideline – Second Edition. EP9-A2. 
 
 
 
 
 
 
 
 
 
 

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Version 3: 18 July 2016 
 
 
 
Purpose 
This protocol is designed to investigate how Allegro results compare to a recognized central laboratory 
reference method. These studies are important in order to establish the relationship between the Allegro 
System and the reference method with respect to statistical correlation and measurement bias. This protocol is 
designed and recommended for the Allegro System.  
Background 
Good practice dictates that the performance of medical devices such as the Allegro System be verified before 
the conductance of clinical studies. Verification studies include: method comparisons to determine how the 
results of the Allegro Systems compare to a recognized reference method, studies of imprecision to determine 
the reproducibility of the various tests available on the Allegro, and to determine the analytical trueness of the 
Allegro measurements. The protocol below describes how to conduct a method comparison (Allegro vs. 
Reference Method).  
Materials Needed 
 
 Nova Biomedical Allegro 
 Allegro Kidney Function Cartridges 
 Nova Biomedical Allegro Quality Control (QC) Solution 
 Reference Analyzer and reagents 
 2mL plastic tubes for decanting urine samples 
 Plastic 2mL disposable pipettes 
 Full range of adjustable pipettes and tips 
 If required appropriate Spiking stock solutions 
 
Important Factors to Consider 
Tests: Urine Creatinine and urine Albumin 
Sample Type: Fresh random urine 
Sample Size: 25µL 
Time per UABCR: 7 mins 
Linear Range of Operation: 15-500 mg/dL (13.0-44.2 mmol/L) 
Linear Range of Operation for Urine Albumin: 3-300 mg/L 
 
Expected Normal Values 
Urine Creatinine: For a random urine sample, >12 Years Male 20-370 mg/dL (1.8- 32.7 mmol/L), >12 Years 
Female 20 – 320 mg/dL (1.8-28.3 mmol/L) 
Urine Albumin: For a random urine sample, normal values are typically 0 to 30 mg/L 
 
 

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Control Levels: 
The Allegro supports two levels of controls.  The following are suggested ranges for each parameter and level: 
Urine Albumin and Creatinine: 
Low Level Target: 
 
Urine Albumin ~50 mg/L 
 
Creatinine ~85mg/dL (~8 mmol/L) 
High Level Target: 
 
Urine Albumin ~180 mg/L 
 
Creatinine ~220 mg/dL (~20 mmol/L) 
 
Please note that urine creatinine and urine Albumin are processed at the same time and can be run in both 
channels 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Version 3: 18 July 2016 
 
 
 
 
Procedure 
1. Fill out the requested information (Allegro serial numbers, cartridge lot numbers, QC lot numbers, QC 
ranges) on the provided Data Sheets (see Appendix).   
 
2. Perform quality control runs on the Allegro being evaluated and the reference analyzer. Record the 
results for all systems on the Data Sheet and make sure the results are within the manufacturer’s 
control range. Proceed with testing only if all control results are within range.  
 
3. 40-50 fresh (within 24hrs) discarded random urine samples should be collected and run in duplicate on 
Allegro.  Again spiking with creatinine spiking solutions and human Albumin may be required to cover 
the measuring range.  
 
4. Ensure adequate mixing before analysis. 
 
5. Samples should be analysed promptly on the reference analyser.  
 
7. Cartridges are stored at 4° each cartridge requires 10-15 mins warm up time so consider this requirement 
when planning study needs. 
 
8. Analyse all samples in duplicate on the Allegro using the same channel to reduce variability 
 
9. Collect, Review and collate data.   
 
10. Total Analysis Time: 7 mins x 80-100 =560 -700 mins. Approx. 9.3 – 11.6 hrs plus preparation time 
 
11. To reduce analysis time consider running samples in singles or perhaps less samples in duplicate whichever 
best suits site 
 
 
Data Analysis 
As part of this study paired urine creatinine and urine Albumin measurements will be collected. Analysis of 
accuracy is based on pairing individual results from the Allegro with the corresponding reference analyser 
measurement and calculating the difference (bias). Scatter plots and linear regression analysis are appropriate 
tools to assess the relationship between the Allegro results and the reference analyzer. Moreover Bland-Altman 
plots can be used to assess the bias relative to the range of urine creatinine and urine Albumin concentrations 
studied. Investigators are referred to organizations such as CLSI and the International Standards Organization 
(ISO) which offer published acceptance criteria [1, 2]. The validity of the conclusions is dependent on 
experimental design, chiefly number of samples, the distribution of results, and the calibration of the reference 
analyzer.  

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Data Review 
This data will be summarized in a summary report which will also include an assessment of precision. This 
report should be reviewed and signed off by the study site principal investigator. Results of this laboratory 
evaluation can be used to document verification of the Allegro System performance prior to the conductance of 
clinical studies.   
 
References 
 
1. Clinical & Laboratory Standards Institute (CLSI). Method Comparison and Bias Estimation Using Patient  
       Samples; Approved Guideline – Second Edition. EP9-A2. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Appendix 4 
 
Method Comparison Data Collection Sheet (Case Report Form)                                                              Page 1 of 3 
 
Study Site: _____________________________ Date of Study: _____________________ 
System Operator(s): ____________________________________________________________ 
 
Specimen Type (Urine) ______________________________________ 
Reference Analyzer ______________________ 
 
 
System Name 
 
 
 
 
 
System Serial 
# 
 
 
 
 
 
Cartridge Lot 
# 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
High QC 
Range:  
 
 
 
 
 
Specimen ID 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Reference 
Analyzer 
Result Urine 
Creat mg/dL 
or mmol/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Reference 
Analyser 
Result Urine 
Albumin mg/L 
or g/L 
1 
 
 
 
 
 
 
2 
 
 
 
 
 
 
3 
 
 
 
 
 
 
4 
 
 
 
 
 
 
5 
 
 
 
 
 
 
6 
 
 
 
 
 
 
7 
 
 
 
 
 
 
8 
 
 
 
 
 
 
9 
 
 
 
 
 
 
10 
 
 
 
 
 
 
11 
 
 
 
 
 
 
12 
 
 
 
 
 
 
13 
 
 
 
 
 
 
14 
 
 
 
 
 
 
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18 
 
 
 
 
 
 
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Method Comparison Data Collection Sheet (Case Report Form)                                                              Page 2 of 3 
 
 
System Name 
 
 
 
 
 
System Serial 
# 
 
 
 
 
 
Cart Lot # 
 
 
 
 
 
Specimen ID 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Reference 
Analyzer 
Result Urine 
Creat mg/dL 
or mmol/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Reference 
Analyser 
Result Urine 
Albumin mg/L 
or g/L 
31 
 
 
 
 
 
 
32 
 
 
 
 
 
 
33 
 
 
 
 
 
 
34 
 
 
 
 
 
 
35 
 
 
 
 
 
 
36 
 
 
 
 
 
 
37 
 
 
 
 
 
 
38 
 
 
 
 
 
 
39 
 
 
 
 
 
 
40 
 
 
 
 
 
 
41 
 
 
 
 
 
 
42 
 
 
 
 
 
 
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Method Comparison Data Collection Sheet (Case Report Form)   
 
 
 
Page 3 of 3 
 
 
 
System Name 
 
 
 
 
 
System Serial 
# 
 
 
 
 
 
Cart Lot # 
 
 
 
 
 
Specimen ID 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Urine Creat 
Result 
mg/dL or 
mmol/L 
Reference 
Analyzer 
Result Urine 
Creat mg/dL 
or mmol/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Urine 
Albumin 
Result 
mg/L or g/L 
Reference 
Analyser 
Result Urine 
Albumin mg/L 
or g/L 
73 
 
 
 
 
 
 
74 
 
 
 
 
 
 
75 
 
 
 
 
 
 
76 
 
 
 
 
 
 
77 
 
 
 
 
 
 
78 
 
 
 
 
 
 
79 
 
 
 
 
 
 
80 
 
 
 
 
 
 
81 
 
 
 
 
 
 
82 
 
 
 
 
 
 
83 
 
 
 
 
 
 
84 
 
 
 
 
 
 
85 
 
 
 
 
 
 
86 
 
 
 
 
 
 
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100 
 
 
 
 
 
 
 
Name of Person Collecting Data: _______________________________ 
Signature of Person Collecting Data: ____________________________ 
Date: ________________________ 
 
 

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1. Urine Creatinine and Urine Albumin 
Summary Report Template 
 
 
 
 
 
 

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Urine Creatinine and Urine Albumin Summary Report  
Date:  
Report Author:  
Study Site:  
Study Site Principal Investigator/Department:  
Materials and Methods 
See Protocols I-IV. Any deviations from protocol should be noted here.  
Summary of Results 
I. Precision Studies 
Table 1. Urine Creatinine Within-Day Precision for Allegro 
 
 
Low Urine 
Creat 
Mid Urine 
Creat 
High Urine 
Creat 
Mean mg/dL or mmol/L 
 
 
 
Standard Deviation 
 
 
 
%CV 
 
 
 
N 
 
 
 
Matrix: Urine 
 
 
 
 
 
Table 2. Urine Albumin Within-Day Precision for Allegro 
 
 
Low Urine 
Urine 
Albumin 
Mid Urine 
Urine 
Albumin 
High Urine 
Urine 
Albumin 
Mean mg/L or g/L 
 
 
 
Standard Deviation 
 
 
 
%CV 
 
 
 
N 
 
 
 
Matrix: Urine 
 
 
 
 
 

29 
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Table 3. Urine Creatinine Between-Day Precision for Allegro 
 
 
Low Urine 
Creat 
Mid Urine 
Creat 
High Urine 
Creat 
Mean mg/dL or mmol/L 
 
 
 
Standard Deviation 
 
 
 
%CV 
 
 
 
N 
 
 
 
Matrix: Urine 
 
 
 
 
Table 4. Urine Albumin Between-Day Precision for Allegro 
 
 
Low Urine 
Urine 
Albumin 
Mid Urine 
Urine 
Albumin 
High Urine 
Urine 
Albumin 
Mean mg/L or g/L 
 
 
 
Standard Deviation 
 
 
 
%CV 
 
 
 
N 
 
 
 
Matrix: Urine 
 
 
 
 
 
II. Method Comparison  
Table 7. Summary of Linear Regression Analysis Results Urine creatinine and Urine Albumin for the Allegro  
 
 
 
 
Urine Creat 
Urine Albumin 
Slope 
 
 
y-intercept (mg/dL) 
 
 
R2 
 
 
N 
 
 
Urine Creat Range mg/dL or mmol/L 
 
 
Urine Albumin Range mg/L or g/L 
 
 
 
Table 8. Summary of Bland-Altman Results 
 
Urine Creat 
Urine Albumin 

30 
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Mean Bias (mg/dL) 
 
 
95% Limits of Agreement 
 
 
         Lower 
 
 
         Upper 
 
 
Urine Creat Range mg/dL or mmol/L 
 
 
Urine Albumin Range mg/L or gl/L 
 
 
 
 
 
Summary Findings 
Allegro demonstrated …. 
 
 
This report has been reviewed and approved by: 
 
Signature of Study Site Principal Investigator                                                                               Date 
 
 
 
Signature of MASA Director                                                                                                              Date 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

31 
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2. HbA1c 
 
 
 
 
 
I. 
Within Day Precision Study Protocol 
Protocol based on: 
 
Clinical & Laboratory Standards Institute (CLSI). Evaluation of Precision Performance of Quantitative 
Measurement Methods; Approved Guideline – Second Edition. EP5-A2. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Purpose 
This protocol is recommended and designed by Nova Biomedical to investigate the within-day precision of 
whole blood HbA1c measurements on the Allegro system. 
Materials Needed 
 
 Nova Biomedical Allegro  
 Allegro Glycaemic Control Cartridges 
 Nova Biomedical Allegro Quality Control (QC) Solution 
 Reference Analyzer and reagents 
 2mL plastic microcentrifuge tubes for decanting EDTA samples 
 Plastic 2mL disposable pipettes 
 Full range of adjustable pipettes and tips 

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 Appropriate Spiking stock solutions 
 
Important Factors to Consider 
Tests: HbA1c 
Sample Type: 
 Capillary blood (from finger stick) 
 Fresh venous whole blood with anticoagulants ethylenediamine tetra-acetic acid (EDTA); Li heparin; Na 
heparin 
Sample Size: 1.5 µL 
Time per HbA1c: 6.5 mins 
Linear Range of Operation for HbA1c:  4-15% (20.2 - 140.4 mmol/L) 
 
Potential Interferences 
HbA1c measurement is not affected by Hb variants: HbAc, HbAD, HbAE, HbF, HbAG and HbAS 
 
Control Levels: 
The Allegro supports two levels of controls.  The following are suggested ranges for each parameter and level: 
HbA1c: 
Low Level Target: 
~5% (31.1 mmol/mol) 
High Level Target: 
~10% (85.5 mmol/mol) 
Normal Values 
HbA1c: - A normal non-diabetic HbA1c is <5.5% (36mmol/mol). In diabetes about 6.5% (48 mmol/mol) is good. 
 
 
 
 
Procedure 
1. Fill out the requested information (Allegro serial numbers, cartridge lot numbers, QC lot numbers, QC 
ranges) on the provided Data Sheets (see Appendix).   
 
2. Perform quality control runs on the Allegro being evaluated and the reference analyzer. Record the 
results for all systems on the Data Sheet and make sure the results are within the manufacturer’s 
control range. Proceed with testing only if all control results are within range.  
 
3. Using aliquoted venous EDTA whole blood identify three levels low, medium and high.  
 
4. Suggested concentrations for low, medium and high samples: - 
 
 
 
 
HbA1c  
 
Low Target 
 
 
~5% (31.1 mmol/mol)  
 
 
Medium Target 
 
~7% (53 mmol/mol)  
 

33 
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High Target 
 
 
~10% (85.8 mmol/mol) 
 
 
5. Ensure adequate mixing before analysis. 
 
6. Cartridges are stored at 4° each cartridge the HbA1c cartridge can be used cold 
 
7. Run each Level 10 times on Allegro, the HbA1c cartridge can only be run in the 600nm channel which is on the 
right hand side of the analyser.  
 
8. Collect, review and collate data.   
 
Total Analysis Time: 6.25 mins x 10 = 62.5 mins per level. 187.5 mins for all levels. Approx. 3.15 hrs plus preparation 
time 
 
Data Analysis 
The within-run precision of the Allegro be assessed by calculating the % coefficient of variation (%CV) from the 
mean and standard deviation from the replicate measurements. Acceptability criteria are determined by the 
medical director and/or institution. Investigators are referred to organizations such as CLSI and the 
International Standards Organization (ISO) which offer published acceptance criteria [4].  
 
Data Review 
This data will be summarized in a final report which will also include an assessment of the accuracy. This report 
should be reviewed by the study site principal investigator. Results of this laboratory evaluation can be used to 
document verification of Allegro performance prior to the conductance of clinical studies. 
 
 
References 
 
1.  Clinical & Laboratory Standards Institute (CLSI). Evaluation of Precision Performance of Quantitative  
        Measurement Methods; Approved Guideline – Second Edition. EP5-A2. 
 
 
 
 
 
 
 
 
 
 
 
 

34 
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Appendix 1 
 
Within-Day Precision Study Data Collection Sheet                                                                                   
 
Hospital Name: _____________________________ Date of Study: _____________________ 
System Operator(s): ____________________________________________________________ 
Reference Analyzer ______________________ 
 
Level 1 HbA1c 
 
Reference Analyzer Result % mmol/mol__________ 
 
 
System Name 
 
 
 
 
 
 
 
System Serial # 
 
 
 
 
 
 
 
Cartridge Lot # 
 
 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
 
 
Replicate # 
HbA1c 
 
 
 
 
 
 

35 
Version 3: 18 July 2016 
 
% or mmol/Mol 
1 
 
 
 
 
 
 
 
2 
 
 
 
 
 
 
 
3 
 
 
 
 
 
 
 
4 
 
 
 
 
 
 
 
5 
 
 
 
 
 
 
 
6 
 
 
 
 
 
 
 
7 
 
 
 
 
 
 
 
8 
 
 
 
 
 
 
 
9 
 
 
 
 
 
 
 
10 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Level 2 HbA1c 
 
Reference Analyzer Result % or mmol/mol __________ 
 
 
System Name 
 
 
 
 
 
 
 
System Serial # 
 
 
 
 
 
 
 
Cartridge Lot # 
 
 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
 
 
Replicate # 
HbA1c 
% or mmol/mol 
 
 
 
 
 
 
1 
 
 
 
 
 
 
 
2 
 
 
 
 
 
 
 
3 
 
 
 
 
 
 
 
4 
 
 
 
 
 
 
 
5 
 
 
 
 
 
 
 
6 
 
 
 
 
 
 
 
7 
 
 
 
 
 
 
 
8 
 
 
 
 
 
 
 

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9 
 
 
 
 
 
 
 
10 
 
 
 
 
 
 
 
 
 
Level 3 HbA1c 
 
Reference Analyzer Result % or mmol/mol__________ 
 
 
System Name 
 
 
 
 
 
 
 
System Serial # 
 
 
 
 
 
 
 
Cartridge Lot # 
 
 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
 
 
Replicate # 
HbA1c 
% or mmol/mol 
 
 
 
 
 
 
1 
 
 
 
 
 
 
 
2 
 
 
 
 
 
 
 
3 
 
 
 
 
 
 
 
4 
 
 
 
 
 
 
 
5 
 
 
 
 
 
 
 
6 
 
 
 
 
 
 
 
7 
 
 
 
 
 
 
 
8 
 
 
 
 
 
 
 
9 
 
 
 
 
 
 
 
10 
 
 
 
 
 
 
 
 
Name of Person Collecting Data: _______________________________ 
Signature of Person Collecting Data: ____________________________ 
Date: ________________________ 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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2. HbA1c 
 
II. Between-Day Precision Study Protocol 
 
Protocol based on: 
Clinical & Laboratory Standards Institute (CLSI). Evaluation of Precision Performance of Quantitative 
Measurement Methods; Approved Guideline – Second Edition. EP5-A2. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Purpose 
This protocol is recommended and designed by Nova Biomedical to investigate the between-day precision of 
whole blood HbA1c measurements on the Allegro system. 
Materials Needed 
 
 Nova Biomedical Allegro 
 Allegro Glycemic Control Cartridges 
 Nova Biomedical Allegro Quality Control (QC) Solution 
 Reference Analyzer and reagents 
 2mL plastic microcentrifuge tubes for decanting EDTA samples 
 Plastic 2mL disposable pipettes 
 Full range of adjustable pipettes and tips 

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Version 3: 18 July 2016 
 
 If required appropriate Spiking stock solutions 
 
Important Factors to Consider 
Tests: HbA1c 
Sample Type:  
 Capillary blood (from finger stick) 
 Fresh venous whole blood with anticoagulants ethylenediamine tetra-acetic acid (EDTA); Li heparin; Na 
heparin 
Sample Size: 1.5 µL 
Time per HbA1c: 6.5 mins 
Linear Range of Operation for HbA1c:  4-15% (20.2 - 140.4 mmol/L) 
 
Potential Interferences 
HbA1c measurement is not affected by Hb variants: HbAc, HbAD, HbAE, HbF, HbAG and HbAS 
 
Control Levels: 
The Allegro supports two levels of controls.  The following are suggested ranges for each parameter and level: 
HbA1c: 
Low Level Target: 
~5% (~31.1 mmol/mol) 
High Level Target: 
~10% (~85.8 mmol/mol) 
Normal Values 
HbA1c:- A normal non-diabetic HbA1c is <5.5% (36mmol/mol). In diabetes about 6.5% (48 mmol/mol) is good. 
 
 
 
 
Procedure 
1. Fill out the requested information (Allegro serial numbers, cartridge lot numbers, QC lot numbers, QC 
ranges) on the provided Data Sheets (see Appendix).   
 
2. Perform quality control runs on the Allegro being evaluated and the reference analyzer. Analyse 
controls on Allegro every day that testing is carried out, 2 runs per day would be performed one in the 
morning and the other in the afternoon so QC would need to be performed prior to each run, make sure 
results are within specification before proceeding. Record the results for all systems on the Data Sheet 
and make sure the results are within the manufacturer’s control range. Proceed with testing only if all 
control results are within range.  
 

39 
Version 3: 18 July 2016 
 
3. Because of the logistical difficulty of using whole blood over a 2-week period the suggestion here is to 
use the Control Solutions, these provide only 2 levels, but these should be aliquoted into 40 x 50µL 
aliquots and frozen to reduce variables.  
 
4. A freshly aliquot will be thawed, mixed and analysed every morning and afternoon in duplicate for 10 
consecutive working days. 
 
6. Cartridges are stored at 4° each cartridge the HbA1c cartridge can be used cold 
 
7. Run each Level in duplicate on Allegro, the HbA1c cartridge can only be run in the 600nm channel which is on 
the right hand side of the analyser. 
 
8. Collect, review and collate data 
Total Analysis Time: 6.25 mins x 4 =25 mins in morning, 6.25 mins x 4 = 25 mins in afternoon. Approx. 50 mins 
plus preparation time   
Data Analysis 
The between-day precision of the Allegro be assessed by calculating the % coefficient of variation (%CV) from 
the mean and standard deviation from the replicate measurements. Acceptability criteria are determined by 
the medical director and/or institution. Investigators are referred to organizations such as CLSI and the 
International Standards Organization (ISO) which offer published acceptance criteria [1].  
 
Data Review 
This data will be summarized in a final report which will also include an assessment of the accuracy. This report 
should be reviewed by the study site principal investigator. Results of this laboratory evaluation can be used to 
document verification of Allegro performance prior to the conductance of clinical studies. 
 
References 
 
1.  Clinical & Laboratory Standards Institute (CLSI). Evaluation of Precision Performance of Quantitative  
        Measurement Methods; Approved Guideline – Second Edition. EP5-A2. 
 
Appendix 2 
 
Between-Day Precision Study Data Collection Sheet                                                                                   
 
Hospital Name: _____________________________ Date of Study: _____________________ 
System Operator(s): ____________________________________________________________ 
Reference Analyzer ______________________ 
 
Level 1 HbA1c 
 
Morning 
Reference Analyzer Result % or mmol/mol __________ 
Afternoon 

40 
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Reference Analyzer Result % or mmol/mol __________ 
 
 
 
System Name 
 
 
 
 
System Serial # 
 
 
 
 
Cartridge Lot # 
 
 
 
 
Low QC 
Range:  
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
Morning 
Afternoon 
 
Replicate # 
HbA1c Result 
% or 
mmol/mol 
HbA1c Result 
% or 
mmol/mol 
HbA1c Result 
% or 
mmol/mol 
HbA1c Result 
% or 
mmol/mol 
Day 1 
 
 
 
 
Day 2 
 
 
 
 
Day 3 
 
 
 
 
Day 4 
 
 
 
 
Day 5 
 
 
 
 
Day 6 
 
 
 
 
Day 7 
 
 
 
 
Day 8 
 
 
 
 
Day 9 
 
 
 
 
Day 10 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Level 2 HbA1c 
 
Morning 
Reference Analyzer Result % or mmol/mol __________ 
Afternoon 
Reference Analyzer Result % or mmol/mol __________ 
 
 
System Name 
 
 
 
 
System Serial # 
 
 
 
 
Cartridge Lot # 
 
 
 
 
Low QC 
Range:  
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
Morning 
Afternoon 

41 
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Replicate # 
HbA1c Result 
% or 
mmol/mol 
HbA1c Result 
% or 
mmol/mol 
HbA1c Result 
% or 
mmol/mol 
HbA1c Result 
% or 
mmol/mol 
Day 1 
 
 
 
 
Day 2 
 
 
 
 
Day 3 
 
 
 
 
Day 4 
 
 
 
 
Day 5 
 
 
 
 
Day 6 
 
 
 
 
Day 7 
 
 
 
 
Day 8 
 
 
 
 
Day 9 
 
 
 
 
Day 10 
 
 
 
 
 
Name of Person Collecting Data: _______________________________ 
Signature of Person Collecting Data: ____________________________ 
Date: ________________________ 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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2. HbA1c 
 
 
 
 
 
III. 
Method Comparison Study Protocol 
 
Protocol based on the following guidance documents: 
 
Clinical & Laboratory Standards Institute (CLSI). Method Comparison and Bias Estimation Using Patient Samples; 
Approved Guideline – Second Edition. EP9-A2. 
 
 
 
 
 
 
 
 
 
 

43 
Version 3: 18 July 2016 
 
Purpose 
This protocol is designed to investigate how Allegro results compare to a recognized central laboratory 
reference method. These studies are important in order to establish the relationship between the Allegro 
System and the reference method with respect to statistical correlation and measurement bias. This protocol is 
designed and recommended for the Allegro System.  
Background 
Good practice dictates that the performance of medical devices such as the Allegro System be verified before 
the conductance of clinical studies. Verification studies include: method comparisons to determine how the 
results of the Allegro Systems compare to a recognized reference method, studies of imprecision to determine 
the reproducibility of the various tests available on the Allegro to determine the analytical trueness of the 
Allegro measurements. The protocol below describes how to conduct a method comparison (Allegro vs. 
Reference Analyser).  
Materials Needed 
 
 Nova Biomedical Allegro 
 Allegro Glycaemic Control Cartridges 
 Nova Biomedical Allegro Quality Control (QC) Solution 
 Reference Analyzer and reagents 
 2mL plastic microcentrifuge tubes for decanting EDTA samples 
 Plastic 2mL disposable pipettes 
 Full range of adjustable pipettes and tips 
 If required appropriate Spiking stock solutions 
 
Important Factors to Consider 
Tests: HbA1c 
Sample Type:  
 Capillary blood (from finger stick) 
 Fresh venous whole blood with anticoagulants ethylenediamine tetra-acetic acid (EDTA); Li heparin; Na 
heparin 
Sample Size: 1.5 µL 
Time per HbA1c: 6.25 mins 
Linear Range of Operation for HbA1c:  4-15% (20.2 - 140.4 mmol/L) 
 
Potential Interferences 
HbA1c measurement is not affected by Hb variants: HbAc, HbAD, HbAE, HbF, HbAG and HbAS 
 
Control Levels: 
The Allegro supports two levels of controls.  The following are suggested ranges for each parameter and level: 

44 
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HbA1c: 
Low Level Target: 
~5% (~31.1 mmol/mol) 
High Level Target: 
~10% (~85.5 mmol/mol) 
 
Normal Values 
HbA1c:- A normal non-diabetic HbA1c is <5.5% (36mmol/mol). In diabetes about 6.5% (48 mmol/mol) is good. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

45 
Version 3: 18 July 2016 
 
Procedure 
1. Fill out the requested information (Allegro serial numbers, cartridge lot numbers, QC lot numbers, QC 
ranges) on the provided Data Sheets (see Appendix).   
 
2. Perform quality control runs on the Allegro being evaluated and the reference analyzer. Record the 
results for all systems on the Data Sheet and make sure the results are within the manufacturer’s 
control range. Proceed with testing only if all control results are within range.  
 
3. Select 40-50 fresh discarded HbA1c EDTA samples that cover the measuring range if possible. 
 
4. Ensure adequate mixing before analysis. 
 
5.  Cartridges are stored at 4° each cartridge the HbA1c cartridge can be used cold 
 
6. Run all samples in duplicate on Allegro following analysis on the reference analyser. Cartridges Allegro, the 
HbA1c cartridge can only be run in the 600nm channel which is on the right hand side of the analyser.  
 
7. Collect, Review and collate data.   
 
8. Total Analysis Time: 6.25 mins x 80 - 100 = 500 - 625 mins. Approx. 8.3 – 10.4 hrs plus preparation time 
 
9. To reduce analysis time consider running samples in singles or perhaps less samples in duplicate whichever 
best suits site 
 
Data Analysis 
As part of this study paired HbA1c measurements will be collected. Analysis of accuracy is based on pairing 
individual results from the Allegro with the corresponding reference analyser measurement and calculating the 
difference (bias). Scatter plots and linear regression analysis are appropriate tools to assess the relationship 
between the Allegro results and the reference analyzer. Moreover, Bland-Altman plots can be used to assess 
the bias relative to the range studied. Investigators are referred to organizations such as CLSI and the 
International Standards Organization (ISO) which offer published acceptance criteria [2]. The validity of the 
conclusions is dependent on experimental design, chiefly number of samples, the distribution of results, and 
the calibration of the reference analyzer.  
 
Data Review 
This data will be summarized in a summary report which will also include an assessment of precision. This 
report should be reviewed and signed off by the study site principal investigator. Results of this laboratory 
evaluation can be used to document verification of the Allegro System performance prior to the conductance of 
clinical studies.   
References 
 
1. Clinical & Laboratory Standards Institute (CLSI). Method Comparison and Bias Estimation Using Patient  
       Samples; Approved Guideline – Second Edition. EP9-A2. 

46 
Version 3: 18 July 2016 
 
Appendix 4 
 
Method Comparison Data Collection Sheet (Case Report Form)                                                            Page 1 of 3 
 
Study Site: _____________________________ Date of Study: _____________________ 
System Operator(s): ____________________________________________________________ 
 
Specimen Type (Venous EDTA Whole Blood) ______________________________________ 
Reference Analyzer ______________________ 
 
 
System Name 
 
 
System Serial #  
 
Cartridge Lot # 
 
 
Low QC 
Range:  
 
 
High QC 
Range:  
 
 
Specimen ID 
Reference 
Analyzer Result % 
or mmol/mol 
HbA1c result 
% or 
mmol/mol 
HbA1c result 
% or 
mmol/mol 
1 
 
 
 
2 
 
 
 
3 
 
 
 
4 
 
 
 
5 
 
 
 
6 
 
 
 
7 
 
 
 
8 
 
 
 
9 
 
 
 
10 
 
 
 
11 
 
 
 
12 
 
 
 
13 
 
 
 
14 
 
 
 
15 
 
 
 
16 
 
 
 
17 
 
 
 
18 
 
 
 
19 
 
 
 
20 
 
 
 
21 
 
 
 
22 
 
 
 
23 
 
 
 
24 
 
 
 
25 
 
 
 
26 
 
 
 
27 
 
 
 
28 
 
 
 
29 
 
 
 
30 
 
 
 
 
 
 
 

47 
Version 3: 18 July 2016 
 
Method Comparison Data Collection Sheet (Case Report Form)                                                             Page 2 of 3 
 
 
System Name 
 
 
System Serial #  
 
Cart Lot # 
 
 
Specimen ID 
Reference 
Analyzer Result % 
or mmol/mol 
HbA1c result 
% or 
mmol/mol 
HbA1c result 
% or 
mmol/mol 
31 
 
 
 
32 
 
 
 
33 
 
 
 
34 
 
 
 
35 
 
 
 
36 
 
 
 
37 
 
 
 
38 
 
 
 
39 
 
 
 
40 
 
 
 
41 
 
 
 
42 
 
 
 
43 
 
 
 
44 
 
 
 
45 
 
 
 
46 
 
 
 
47 
 
 
 
48 
 
 
 
48 
 
 
 
49 
 
 
 
50 
 
 
 
51 
 
 
 
52 
 
 
 
53 
 
 
 
54 
 
 
 
55 
 
 
 
56 
 
 
 
57 
 
 
 
58 
 
 
 
59 
 
 
 
60 
 
 
 
61 
 
 
 
62 
 
 
 
63 
 
 
 
64 
 
 
 
65 
 
 
 
66 
 
 
 
67 
 
 
 
68 
 
 
 
69 
 
 
 
70 
 
 
 
71 
 
 
 
72 
 
 
 
73 
 
 
 

48 
Version 3: 18 July 2016 
 
Method Comparison Data Collection Sheet (Case Report Form)   
 
 
 
Page 3 of 3 
 
 
 
System Name 
 
 
System Serial #  
 
Cart Lot # 
 
 
Specimen ID 
Reference 
Analyzer Result % 
or mmol/mol 
HbA1c result 
% or 
mmol/mol 
HbA1c result 
% or 
mmol/mol 
74 
 
 
 
75 
 
 
 
76 
 
 
 
77 
 
 
 
78 
 
 
 
79 
 
 
 
80 
 
 
 
81 
 
 
 
82 
 
 
 
83 
 
 
 
84 
 
 
 
85 
 
 
 
86 
 
 
 
87 
 
 
 
88 
 
 
 
89 
 
 
 
90 
 
 
 
91 
 
 
 
92 
 
 
 
93 
 
 
 
94 
 
 
 
95 
 
 
 
96 
 
 
 
97 
 
 
 
98 
 
 
 
99 
 
 
 
100 
 
 
 
 
Name of Person Collecting Data: _______________________________ 
Signature of Person Collecting Data: ____________________________ 
Date: ________________________ 
 
 
 
 
 
 
 
 
 
 
 

49 
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2. HbA1c 
Summary Report Template 
 
 
 
 
 
 
 
 
 
 

50 
Version 3: 18 July 2016 
 
HbA1c Summary Report  
Date:  
Report Author:  
Study Site:  
Study Site Principal Investigator/Department:  
Materials and Methods 
See Protocols I-IV. Any deviations from protocol should be noted here.  
Summary of Results 
I. Precision Studies 
Table 1. HbA1c Within-Day Precision for Allegro 
 
 
Low HbA1c 
Mid HbA1c 
High HbA1c 
Mean % or mol/mol 
 
 
 
Standard Deviation 
 
 
 
%CV 
 
 
 
N 
 
 
 
Matrix 
 
 
 
 
Table 2. HbA1c Between-Day Precision for Allegro 
 
 
Low HbA1c 
Mid HbA1c 
High HbA1c 
Mean % or mol/mol 
 
 
 
Standard Deviation 
 
 
 
%CV 
 
 
 
N 
 
 
 
Matrix 
 
 
 
 
 
 
 
 
 
 
 

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Version 3: 18 July 2016 
 
II. Method Comparison 
Table 5. Summary of Linear Regression Analysis Results HbA1C for Allegro 
 
 
 
HbA1c 
 
Slope 
 
y-intercept (% or mmol/mol)) 
 
R2 
 
N 
 
HbA1c Range % or mmol/mol 
 
 
 
 
 
Table 6. Summary of Bland-Altman Results 
 
 
HbA1c 
Mean Bias (% or mmol/mol) 
 
95% Limits of Agreement 
 
         Lower 
 
         Upper 
 
HbA1c (% or mmol/mol) 
 
 
 
 
 
 
 
Summary Findings 
Allegro demonstrated …. 
 
 
This report has been reviewed and approved by: 

52 
Version 3: 18 July 2016 
 
 
Signature of Study Site Principal Investigator                                                                               Date 
 
 
Signature of MASA Director                                                                                                              Date 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
3. Total Cholesterol, Triglyceride and HDL Cholesterol 
 
 
 
 
 
I. 
Within Day Precision Study Protocol 
Protocol based on: 
 
Clinical & Laboratory Standards Institute (CLSI). Evaluation of Precision Performance of Quantitative 
Measurement Methods; Approved Guideline – Second Edition. EP5-A2. 

53 
Version 3: 18 July 2016 
 
 
 
 
 

54 
Version 3: 18 July 2016 
 
Purpose 
This protocol is recommended and designed by Nova Biomedical to investigate the within day precision of 
whole blood total cholesterol, triglycerides and HDL cholesterol measurements on the Allegro system. 
Materials Needed 
 
 Nova Biomedical Allegro  
 Allegro Lipid Risk Cartridges 
 Nova Biomedical Allegro Quality Control (QC) Solution 
 Reference Analyzer and reagents 
 2mL plastic microcentrifuge tubes for decanting Na Heparin or Li Heparin samples 
 Plastic 2mL disposable pipettes 
 Full range of adjustable pipettes and tips 
 If required appropriate Spiking stock solutions 
 
Important Factors to Consider 
Tests: Total Cholesterol, Triglycerides, HDL Cholesterol 
Sample Type: Subject to change but to date: 
 Capillary whole blood (from finger stick) 
 Venous whole blood with anticoagulants Na heparin; Li heparin 
 Venous plasma with anticoagulants Na heparin; Li heparin 
 Serum 
Sample Size: 5 µL 
Time per Lipid: 10 mins 
Linear Range of Operation for Total Cholesterol: 100-450 mg/dL (2.59-11.64 mmol/L) 
Linear Range of Operation for Triglyceride: 50-600 mg/dL (0.56-6.76 mmol/L) 
Linear Range of Operation for HDL Cholesterol: 20-100 mg/dL (0.52- 2.59 mmol/L) 
 
Please note that all parameters are processed at the same time. 
 
Normal Values: 
Total Cholesterol: Desirable is below 200 mg/dL (<5.17 mmol/L) 
Triglycerides: Desirable is below 150 mg/dL (<1.69 mmol/L) 
HDL Cholesterol: Desirable is 60 mg/dL and above (> 1.55 mg/dL) 
 
 
 
 
 
 
 
 

55 
Version 3: 18 July 2016 
 
 
Control Levels: 
The Allegro supports two levels of controls.  The following are suggested ranges for each parameter and level: 
 
 
    
  Total Cholesterol 
 
Triglycerides  
HDL Cholesterol 
Low Level Target: 
~ 170 mg/dL      
 
 ~150 mg/dL   
~40 mg/dL  
 
 
 
 (~4.40 mmol/L) 
 
(~1.69 mmol/L) 
 
(~1.04 mmol/L) 
 
High Level Target: 
~ 240 mg/dL  
 
~300 mg/dL 
 
~60 mg/dL (1.55 mmol/L) 
 
 
 
(~6.21 mmol/L) 
 
(~3.39 mmol/L) 
 
(~1.55 mmol/L) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Procedure 

56 
Version 3: 18 July 2016 
 
1. Fill out the requested information (Allegro serial numbers, cartridge lot numbers, QC lot numbers, QC 
ranges) on the provided Data Sheets (see Appendix).   
 
2. Perform quality control runs on the Allegro being evaluated and the reference analyzer. Record the 
results for all systems on the Data Sheet and make sure the results are within the manufacturer’s 
control range. Proceed with testing only if all control results are within range.  
 
3. If possible using fresh aliquoted venous Na or Lithium heparin whole blood samples from qualified 
patients identify levels low, medium and high Total Cholesterol, Triglycerides and HDL Cholesterol 
levels.  
 
4. Suggested concentrations for low, medium and high samples: - 
Total Cholesterol 
Triglycerides  
HDL Cholesterol 
Low Level Target:  
~ 150 mg/dL      
 ~100 mg/dL   
~40 mg/dL  
 
 
 
  
(~3.88 mmol/L) 
(~1.13mmol/L) 
(~1.03 mmol/L) 
 
Medium Level Target: 
~ 250 mg/dL  
~200 mg/dL 
 
~60 mg/dL  
 
 
 
 
(~6.47 mmol/L) 
(~2.26 mmol/L) 
(~1.55 mmol/L) 
 
High Level Target:  
~ 400 mg/dL  
~300 mg/dL 
 
~80 mg/dL  
 
 
 
 
(~10.35 mmol/L) 
~3.39 mmol/L) 
(~2.07 mmol/L) 
 
2. Cartridges are stored at 4° each cartridge the HbA1c cartridge can be used cold 
 
3. Cartridges are stored at 4° each cartridge the lipids cartridge is only stable at RT for 1 hour 
 
4. Run each Level 10 times on Allegro, the Lipids cartridge can only be run in the 660nm channel which is on the left 
hand side of the analyser. 
 
6. Collect, review and collate data 
 
Total Analysis time: 10 x 10 mins=100 mins per level. 300 mins for all levels this is just run time. Approx. 5 
hrs. plus preparation time  
 
 
Data Analysis 
The within-run precision of the Allegro be assessed by calculating the % coefficient of variation (%CV) from the 
mean and standard deviation from the replicate measurements. Acceptability criteria are determined by the 
medical director and/or institution. Investigators are referred to organizations such as CLSI and the 
International Standards Organization (ISO) which offer published acceptance criteria [1, 2].  
 
 
Data Review 

57 
Version 3: 18 July 2016 
 
This data will be summarized in a final report which will also include an assessment of the accuracy. This report 
should be reviewed by the study site principal investigator. Results of this laboratory evaluation can be used to 
document verification of Allegro performance prior to the conductance of clinical studies. 
References 
 
1.  Clinical & Laboratory Standards Institute (CLSI). Evaluation of Precision Performance of Quantitative  
        Measurement Methods; Approved Guideline – Second Edition. EP5-A2. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Appendix 1 
 
Within-Day Precision Study Data Collection Sheet                                                                                   

58 
Version 3: 18 July 2016 
 
 
Hospital Name: _____________________________ Date of Study: _____________________ 
System Operator(s): ____________________________________________________________ 
Reference Analyzer ______________________ 
 
Level 1 Total Cholesterol, Triglycerides and HDL Cholesterol 
 
Reference Analyzer Result mg/dL or mmol/L Total Cholesterol __________ 
Reference Analyzer Result mg/dL or mmol/L Triglycerides __________ 
Reference Analyzer Result mg/dL or mmol/L HDL Cholesterol __________ 
 
 
 
System Name 
 
 
 
 
 
 
 
System Serial # 
 
 
 
 
 
 
 
Cartridge Lot # 
 
 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
 
 
Replicate # 
Total 
Cholesterol 
mg/dL or 
mmol/L 
Triglycerides 
mg/dL or 
mmol/L 
HDL Cholesterol 
mg/dL or 
mmol/L 
 
 
 
 
1 
 
 
 
 
 
 
 
2 
 
 
 
 
 
 
 
3 
 
 
 
 
 
 
 
4 
 
 
 
 
 
 
 
5 
 
 
 
 
 
 
 
6 
 
 
 
 
 
 
 
7 
 
 
 
 
 
 
 
8 
 
 
 
 
 
 
 
9 
 
 
 
 
 
 
 
10 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Level 2 Total Cholesterol, Triglycerides and HDL Cholesterol 
 
Reference Analyzer Result mg/dL or mmol/L Total Cholesterol __________ 
Reference Analyzer Result mg/dL or mmol/L Triglycerides __________ 

59 
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Reference Analyzer Result mg/dL or mmol/L HDL Cholesterol __________ 
 
 
System Name 
 
 
 
 
 
 
 
System Serial # 
 
 
 
 
 
 
 
Cartridge Lot # 
 
 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
 
 
Replicate # 
Total 
Cholesterol 
mg/dL or 
mmol/L 
Triglycerides 
mg/dL or 
mmol/L 
HDL Cholesterol 
mg/dL or 
mmol/L 
 
 
 
 
1 
 
 
 
 
 
 
 
2 
 
 
 
 
 
 
 
3 
 
 
 
 
 
 
 
4 
 
 
 
 
 
 
 
5 
 
 
 
 
 
 
 
6 
 
 
 
 
 
 
 
7 
 
 
 
 
 
 
 
8 
 
 
 
 
 
 
 
9 
 
 
 
 
 
 
 
10 
 
 
 
 
 
 
 
 
Level 3 Total Cholesterol, Triglycerides and HDL Cholesterol 
 
Reference Analyzer Result mg/dL or mmol/L Total Cholesterol __________ 
Reference Analyzer Result mg/dL or mmol/L Triglycerides __________ 
Reference Analyzer Result mg/dL or mmol/L HDL Cholesterol __________ 
 
 
System Name 
 
 
 
 
 
 
 
System Serial # 
 
 
 
 
 
 
 
Cart Lot # 
 
 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
 
 
Replicate # 
Total 
Cholesterol 
mg/dL or 
mmol/L 
Triglycerides 
mg/dL or 
mmol/L 
HDL Cholesterol 
mg/dL or 
mmol/L 
 
 
 
 
1 
 
 
 
 
 
 
 
2 
 
 
 
 
 
 
 
3 
 
 
 
 
 
 
 
4 
 
 
 
 
 
 
 
5 
 
 
 
 
 
 
 
6 
 
 
 
 
 
 
 
7 
 
 
 
 
 
 
 
8 
 
 
 
 
 
 
 
9 
 
 
 
 
 
 
 
10 
 
 
 
 
 
 
 
 
Name of Person Collecting Data: _______________________________ 
Signature of Person Collecting Data: ____________________________ 
Date: ________________________ 
 
 

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3. Total Cholesterol, Triglyceride and HDL Cholesterol 
 
II. Between-Day Precision Study Protocol 
 
Protocol based on: 
Clinical & Laboratory Standards Institute (CLSI). Evaluation of Precision Performance of Quantitative 
Measurement Methods; Approved Guideline – Second Edition. EP5-A2. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

61 
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Purpose 
This protocol is recommended and designed by Nova Biomedical to investigate the between-day precision of 
urine creatinine and urine albumin measurements on the Allegro system. 
Materials Needed 
 
 Nova Biomedical Allegro  
 Allegro Lipid Cartridges 
 Nova Biomedical Allegro Lipid Quality Control (QC) Solution 
 Reference Analyzer and reagents 
 2mL plastic microcentrifuge tubes for decanting Na Heparin or Li Heparin samples 
 Plastic 2mL disposable pipettes 
 Full range of adjustable pipettes and tips 
 If required appropriate Spiking stock solutions 
 
Important Factors to Consider 
Tests: Total Cholesterol, Triglycerides, HDL Cholesterol 
Sample Type: Subject to change but to date: 
 Capillary whole blood (from finger stick) 
 Venous whole blood with anticoagulants Na heparin; Li heparin 
 Venous plasma with anticoagulants Na heparin; Li heparin 
 Serum 
Sample Size: 5 µL 
Time per Lipid: 10 mins 
Linear Range of Operation for Total Cholesterol: 100-450 mg/dL (2.59-11.64 mmol/L) 
Linear Range of Operation for Triglyceride: 50-600 mg/dL (0.56-6.76 mmol/L) 
Linear Range of Operation for HDL Cholesterol: 20-100 mg/dL (0.52- 2.59 mmol/L) 
 
Please note that all parameters are processed at the same time. 
 
Normal Values: 
Total Cholesterol: Desirable is below 200 mg/dL (<5.17 mmol/L) 
Triglycerides: Desirable is below 150 mg/dL (<1.69 mmol/L) 
HDL Cholesterol: Desirable is 60 mg/dL and above (> 1.55 mg/dL) 
 
Accuracy Expectations 
Total Cholesterol: 
Target value + 3%   
 
Triglycerides:  
Target value + 5%   
HDL Cholesterol: 
Target value + 5%   
 
 
 

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Version 3: 18 July 2016 
 
Control Levels: 
The Allegro supports two levels of controls.  The following are suggested ranges for each parameter and level: 
 
 
    
  Total Cholesterol 
 
Triglycerides  
HDL Cholesterol 
Low Level Target: 
~ 170 mg/dL      
 
 ~150 mg/dL   
~40 mg/dL  
 
 
 
 (~4.40 mmol/L) 
 
(~1.69 mmol/L) 
 
(~1.04 mmol/L) 
 
High Level Target: 
~ 240mg/dL 
 
 
~300 mg/dL 
 
~60 mg/dL (1.55 mmol/L) 
 
 
 
(~6.21 mmol/L) 
 
(~3.39 mmol/L) 
 
(~1.55 mmol/L) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Procedure 

63 
Version 3: 18 July 2016 
 
1. Fill out the requested information (Allegro serial numbers, cartridge lot numbers, QC lot numbers, QC 
ranges) on the provided Data Sheets (see Appendix).   
 
2. Perform quality control runs on the Allegro being evaluated and the reference analyzer. Analyse 
controls on Allegro every day that testing is carried out, 2 runs per day would be performed one in the 
morning and the other in the afternoon so QC would need to be performed prior to each run, make sure 
results are within specification before proceeding. Record the results for all systems on the Data Sheet 
and make sure the results are within the manufacturer’s control range. Proceed with testing only if all 
control results are within range.  
 
3. Because of the logistical difficulty of using whole blood over a 2-week period the suggestion here is to 
use the Control Solutions, these provide only 2 levels, but these should be aliquoted into 40 x 50µL 
aliquots and frozen to reduce variables.  
 
Note: If Allegro can analyse plasma then the samples used for within day precision can be centrifuged, 
aliquoted into 40 x 50µL microcentrifuge tubes, and frozen. 
 
4. A freshly thawed aliquot will then be analysed every morning and afternoon in duplicate for 10 
consecutive working days 
 
5. Ensure adequate mixing before analysis. 
 
5. Cartridges are stored at 4° each cartridge the lipids cartridge is only stable at RT for 1 hour 
 
6. Run each Level in duplicate on the Allegro, the Lipids cartridge can only be run in the 660nm channel which is on 
the left hand side of the analyser. 
 
6. Collect, review and collate data 
 
Total Analysis Time: 10 mins x 4 = 40 mins in morning 10 mins x 4 = 40 mins in afternoon plus preparation 
time   
Data Analysis 
The between-day precision of the Allegro be assessed by calculating the % coefficient of variation (%CV) from 
the mean and standard deviation from the replicate measurements. Acceptability criteria are determined by 
the medical director and/or institution. Investigators are referred to organizations such as CLSI and the 
International Standards Organization (ISO) which offer published acceptance criteria [1, 2].  
 
 
 
 
 
Data Review 

64 
Version 3: 18 July 2016 
 
This data will be summarized in a final report which will also include an assessment of the accuracy. This report 
should be reviewed by the study site principal investigator. Results of this laboratory evaluation can be used to 
document verification of Allegro performance prior to the conductance of clinical studies. 
References 
 
1.  Clinical & Laboratory Standards Institute (CLSI). Evaluation of Precision Performance of Quantitative  
        Measurement Methods; Approved Guideline – Second Edition. EP5-A2. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Appendix 2 
 
Between-Day Precision Study Data Collection Sheet                                                                                   

65 
Version 3: 18 July 2016 
 
 
Hospital Name: _____________________________ Date of Study: _____________________ 
System Operator(s): ____________________________________________________________ 
Reference Analyzer ______________________ 
 
Level 1 Total Cholesterol, Triglycerides and HDL Cholesterol 
 
Morning 
Reference Analyzer Result mg/dL or mmol/L Total Cholesterol __________ 
Reference Analyzer Result mg/dL or mmol/L Triglycerides __________ 
Reference Analyzer Result mg/dL or mmol/L HDL Cholesterol __________ 
Afternoon  
Reference Analyzer Result mg/dL or mmol/L Total Cholesterol __________ 
Reference Analyzer Result mg/dL or mmol/L Triglycerides __________ 
Reference Analyzer Result mg/dL or mmol/L HDL Cholesterol __________ 
 
 
System Name 
 
 
 
 
 
 
 
 
 
 
 
 
System Serial # 
 
 
 
 
 
 
 
 
 
 
 
 
Cartridge Lot # 
 
 
 
 
 
 
 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
 
 
 
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Morning 
 
 
Afternoon 
 
 
 
Replicate # 
Total Chol 
mg/dL or 
mmol/L 
Total Chol 
mg/dL or 
mmol/L) 
Trigs 
mg/dL 
or 
mmol/L 
Trigs 
mg/dL 
or 
mmol/L 
HDL 
Chol 
mg/dL 
or 
mmol/L 
HDL 
Chol 
mg/dL 
or 
mmol/L 
Total Chol 
mg/dL or 
mmol/L 
Total Chol 
mg/dL or 
mmol/L) 
Trigs 
mg/dL 
or 
mmol/L 
Trigs 
mg/dL 
or 
mmol/L 
HDL 
Chol 
mg/dL 
or 
mmol/L 
HDL 
Chol 
mg/dL 
or 
mmol/L 
Day 1 
 
 
 
 
 
 
 
 
 
 
 
 
Day 2 
 
 
 
 
 
 
 
 
 
 
 
 
Day 3 
 
 
 
 
 
 
 
 
 
 
 
 
Day 4 
 
 
 
 
 
 
 
 
 
 
 
 
Day 5 
 
 
 
 
 
 
 
 
 
 
 
 
Day 6 
 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
 
 
 
 
 
 
 
 
 
 
 
 
Day 8 
 
 
 
 
 
 
 
 
 
 
 
 
Day 9 
 
 
 
 
 
 
 
 
 
 
 
 
Day 10 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Level 2 Total Cholesterol, Triglycerides and HDL Cholesterol 
 
Morning 
Reference Analyzer Result mg/dL or mmol/L Total Cholesterol __________ 

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Reference Analyzer Result mg/dL or mmol/L Triglycerides __________ 
Reference Analyzer Result mg/dL or mmol/L HDL Cholesterol __________ 
Afternoon  
Reference Analyzer Result mg/dL or mmol/L Total Cholesterol __________ 
Reference Analyzer Result mg/dL or mmol/L Triglycerides __________ 
Reference Analyzer Result mg/dL or mmol/L HDL Cholesterol __________ 
 
 
System Name 
 
 
 
 
 
 
 
 
 
 
 
 
System Serial # 
 
 
 
 
 
 
 
 
 
 
 
 
Cartridge Lot # 
 
 
 
 
 
 
 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
 
 
 
 
 
 
 
High QC 
Range: 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Morning 
 
 
Afternoon 
 
 
 
Replicate # 
Total 
Chol 
mg/dL 
or 
mmol/L 
Total 
Chol 
mg/dL or 
mmol/L) 
Trigs 
mg/dL 
or 
mmol/L 
Trigs 
mg/dL 
or 
mmol/L 
HDL 
Chol 
mg/dL 
or 
mmol/L 
HDL 
Chol 
mg/dL 
or 
mmol/L 
Total 
Chol 
mg/dL or 
mmol/L 
Total 
Chol 
mg/dL 
or 
mmol/L) 
Trigs 
mg/dL or 
mmol/L 
Trigs 
mg/dL 
or 
mmol/L 
HDL 
Chol 
mg/dL 
or 
mmol/L 
HDL 
Chol 
mg/dL 
or 
mmol/L 
Day 1 
 
 
 
 
 
 
 
 
 
 
 
 
Day 2 
 
 
 
 
 
 
 
 
 
 
 
 
Day 3 
 
 
 
 
 
 
 
 
 
 
 
 
Day 4 
 
 
 
 
 
 
 
 
 
 
 
 
Day 5 
 
 
 
 
 
 
 
 
 
 
 
 
Day 6 
 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
 
 
 
 
 
 
 
 
 
 
 
 
Day 8 
 
 
 
 
 
 
 
 
 
 
 
 
Day 9 
 
 
 
 
 
 
 
 
 
 
 
 
Day 10 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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3. Total Cholesterol, Triglyceride and HDL Cholesterol 
 
 
 
 
 
III. 
Method Comparison Study Protocol 
 
Protocol based on the following guidance documents: 
 
Clinical & Laboratory Standards Institute (CLSI). Method Comparison and Bias Estimation Using Patient Samples; 
Approved Guideline – Second Edition. EP9-A2. 
 
 
 
 
 
 
 
 
 

68 
Version 3: 18 July 2016 
 
 
 
Purpose 
This protocol is designed to investigate how Allegro results compare to a recognized central laboratory 
reference method. These studies are important in order to establish the relationship between the Allegro 
System and the reference method with respect to statistical correlation and measurement bias. This protocol is 
designed and recommended for the Allegro System.  
Background 
Good practice dictates that the performance of medical devices such as the Allegro System be verified before 
the conductance of clinical studies. Verification studies include: method comparisons to determine how the 
results of the Allegro Systems compare to a recognized reference method, studies of imprecision to determine 
the reproducibility of the various tests available on the Allegro studies to determine the analytical trueness of 
the Allegro measurements. The protocol below describes how to conduct a method comparison (Allegro vs. 
Reference Method).  
Materials Needed 
 
 Nova Biomedical Allegro 
 Allegro Lipid Cartridges 
 Nova Biomedical Allegro Lipid Quality Control (QC) Solution 
 Reference Analyzer and reagents 
 2mL plastic microcentrifuge tubes for decanting Na heparin or Li heparin samples 
 Plastic 2mL disposable pipettes 
 Full range of adjustable pipettes and tips 
 If required appropriate Spiking stock solutions 
 
Important Factors to Consider 
Tests: Total Cholesterol, Triglycerides, HDL Cholesterol 
Sample Type: Subject to change but to date: 
 Capillary whole blood (from finger stick) 
 Venous whole blood with anticoagulants Na heparin; Li heparin 
 Venous plasma with anticoagulants Na heparin; Li heparin 
 Serum 
Sample Size: 5 µL 
Time per Lipid: 10 mins 
Linear Range of Operation for Total Cholesterol: 100-450 mg/dL (2.59-11.64 mmol/L) 
Linear Range of Operation for Triglyceride: 50-600 mg/dL (0.56-6.76 mmol/L) 
Linear Range of Operation for HDL Cholesterol: 20-100 mg/dL (0.52- 2.59 mmol/L) 
 

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Version 3: 18 July 2016 
 
Please note that all parameters are processed at the same time. 
 
Normal Values: 
Total Cholesterol: Desirable is below 200 mg/dL (<5.17 mmol/L) 
Triglycerides: Desirable is below 150 mg/dL (<1.69 mmol/L) 
HDL Cholesterol: Desirable is 60 mg/dL and above (> 1.55 mg/dL) 
 
 
Control Levels: 
The Allegro supports two levels of controls.  The following are suggested ranges for each parameter and level: 
 
 
    
  Total Cholesterol 
 
Triglycerides  
HDL Cholesterol 
Low Level Target: 
~ 170 mg/dL      
 
 ~150 mg/dL   
~40 mg/dL  
 
 
 
 (~4.40 mmol/L) 
 
(~1.69 mmol/L) 
(~1.04 mmol/L) 
 
High Level Target: 
~ 240 mg/dL  
 
~300 mg/dL 
 
~60 mg/dL (1.55 mmol/L) 
 
 
 
(~6.21 mmol/L) 
 
(~3.39 mmol/L) 
(~1.55 mmol/L) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Version 3: 18 July 2016 
 
 
 
Procedure 
1. Fill out the requested information (Allegro serial numbers, cartridge lot numbers, QC lot numbers, QC 
ranges) on the provided Data Sheets (see Appendix).   
 
2. Perform quality control runs on the Allegro being evaluated and the reference analyzer. Record the 
results for all systems on the Data Sheet and make sure the results are within the manufacturer’s 
control range. Proceed with testing only if all control results are within range.  
 
3. 40-50 freshly collected/discarded whole blood venous, Na or Lithium heparin samples should be 
selected, that if possible,  cover the analytical range across all analytes.  Again spiking with appropriate 
spiking solutions may be required to completely cover the analytical range.  
 
4. Ensure adequate mixing before analysis. 
 
5. Analyse all samples in duplicate on the Allegro.  
 
6. Samples should be spun down promptly (plasma) and then be analysed on the reference analyser. 
  
7. Cartridges are stored at 4° each cartridge the lipids cartridge is only stable at RT for 1 hour 
 
8. Run all samples in duplicate Allegro, the Lipids cartridge can only be run in the 660nm channel which is on the 
left hand side of the analyser. 
 
9. Collect, Review and collate data.   
 
10. Total Analysis Time: 10 mins x 80-100 = 800-1000 mins. Approx. 13.3 – 16.6  hrs plus preparation time 
 
11. To reduce analysis time consider running samples in singles or perhaps less samples in duplicate whichever 
best suits site 
Data Analysis 
As part of this study paired total cholesterol, triglycerides and HDL cholesterol measurements will be collected. 
Analysis of accuracy is based on pairing individual results from the Allegro with the corresponding reference 
analyser measurement and calculating the difference (bias). Scatter plots and linear regression analysis are 
appropriate tools to assess the relationship between the Allegro results and the reference analyzer. Moreover, 
Bland-Altman plots can be used to assess the bias relative to the range of urine creatinine and urine Albumin 
concentrations studied. Investigators are referred to organizations such as CLSI and the International Standards 
Organization (ISO) which offer published acceptance criteria [1, 2]. The validity of the conclusions is dependent 
on experimental design, chiefly number of samples, the distribution of results, and the calibration of the 
reference analyzer.  
 

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Data Review 
This data will be summarized in a summary report which will also include an assessment of precision. This 
report should be reviewed and signed off by the study site principal investigator. Results of this laboratory 
evaluation can be used to document verification of the Allegro System performance prior to the conductance of 
clinical studies.   
References 
 
1. Clinical & Laboratory Standards Institute (CLSI). Method Comparison and Bias Estimation Using Patient  
       Samples; Approved Guideline – Second Edition. EP9-A2. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Appendix 4 
 
Method Comparison Data Collection Sheet (Case Report Form)                                                            Page 1 of 3 
 
Study Site: _____________________________ Date of Study: _____________________ 
System Operator(s): ____________________________________________________________ 
 
Specimen Type (Na or Lithium heparin venous blood) ______________________________________ 
Reference Analyzer Result mg/dL or mmol/L Total Cholesterol __________ 
Reference Analyzer Result mg/dL or mmol/L Triglycerides __________ 
Reference Analyzer Result mg/dL or mmol/L HDL Cholesterol __________ 
 
 
System Name  
 
 
 
 
 
 
 
System Serial 
# 
 
 
 
 
 
 
 
 
Cartridge Lot 
# 
 
 
 
 
 
 
 
 
Low QC 
Range:  
 
 
 
 
 
 
 
 
High QC 
Range:  
 
 
 
 
 
 
 
 
Specimen ID 
Total Chol 
mg/dL or 
mmol/L 
Total Chol 
mg/dL or 
mmol/L 
Reference 
Analyzer 
Result Total 
Chol  (mg/dL 
or mmol/L) 
Trigs 
mg/dL or 
mmol/L 
Trigs 
mg/dL/ or 
mmol/L 
Reference 
Analyzer 
Result Trig 
(mg/dL or 
mmol/L) 
HDL Chol 
mg/dL or 
mmol/L 
HDL Chol 
mg/dL or 
mmol/L 
Reference 
Analyzer 
Result HDL 
Chol(mg/dL 
or mmol/L) 
1 
 
 
 
 
 
 
 
 
 
2 
 
 
 
 
 
 
 
 
 
3 
 
 
 
 
 
 
 
 
 
4 
 
 
 
 
 
 
 
 
 
5 
 
 
 
 
 
 
 
 
 
6 
 
 
 
 
 
 
 
 
 
7 
 
 
 
 
 
 
 
 
 
8 
 
 
 
 
 
 
 
 
 
9 
 
 
 
 
 
 
 
 
 
10 
 
 
 
 
 
 
 
 
 
11 
 
 
 
 
 
 
 
 
 
12 
 
 
 
 
 
 
 
 
 
13 
 
 
 
 
 
 
 
 
 
14 
 
 
 
 
 
 
 
 
 
15 
 
 
 
 
 
 
 
 
 
16 
 
 
 
 
 
 
 
 
 
17 
 
 
 
 
 
 
 
 
 
18 
 
 
 
 
 
 
 
 
 
19 
 
 
 
 
 
 
 
 
 
20 
 
 
 
 
 
 
 
 
 
21 
 
 
 
 
 
 
 
 
 
22 
 
 
 
 
 
 
 
 
 
23 
 
 
 
 
 
 
 
 
 
24 
 
 
 
 
 
 
 
 
 
25 
 
 
 
 
 
 
 
 
 

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26 
 
 
 
 
 
 
 
 
 
27 
 
 
 
 
 
 
 
 
 
28 
 
 
 
 
 
 
 
 
 
29 
 
 
 
 
 
 
 
 
 
Method Comparison Data Collection Sheet (Case Report Form)                                                              Page 2 of 3 
 
 
System Name 
 
 
 
 
 
 
 
 
System Serial #  
 
 
 
 
 
 
 
Cart Lot # 
 
 
 
 
 
 
 
 
Specimen ID 
Total Chol 
mg/dL or mmol/L 
Total Chol 
mg/dL or 
mmol/L 
Reference 
Analyzer 
Result Total 
Chol  (mg/dL 
or mmol/L) 
Trigs 
mg/dL or 
mmol/L 
Trigs 
mg/dL/ or 
mmol/L 
Reference 
Analyzer 
Result Trig 
(mg/dL or 
mmol/L) 
HDL Chol 
mg/dL or 
mmol/L 
HDL Chol 
mg/dL or 
mmol/L 
Reference 
Analyzer 
Result HDL 
Chol(mg/dL or 
mmol/L) 
30 
 
 
 
 
 
 
 
 
 
31 
 
 
 
 
 
 
 
 
 
32 
 
 
 
 
 
 
 
 
 
33 
 
 
 
 
 
 
 
 
 
34 
 
 
 
 
 
 
 
 
 
35 
 
 
 
 
 
 
 
 
 
36 
 
 
 
 
 
 
 
 
 
37 
 
 
 
 
 
 
 
 
 
38 
 
 
 
 
 
 
 
 
 
39 
 
 
 
 
 
 
 
 
 
40 
 
 
 
 
 
 
 
 
 
41 
 
 
 
 
 
 
 
 
 
42 
 
 
 
 
 
 
 
 
 
43 
 
 
 
 
 
 
 
 
 
44 
 
 
 
 
 
 
 
 
 
45 
 
 
 
 
 
 
 
 
 
46 
 
 
 
 
 
 
 
 
 
47 
 
 
 
 
 
 
 
 
 
48 
 
 
 
 
 
 
 
 
 
48 
 
 
 
 
 
 
 
 
 
49 
 
 
 
 
 
 
 
 
 
50 
 
 
 
 
 
 
 
 
 
51 
 
 
 
 
 
 
 
 
 
52 
 
 
 
 
 
 
 
 
 
53 
 
 
 
 
 
 
 
 
 
54 
 
 
 
 
 
 
 
 
 
55 
 
 
 
 
 
 
 
 
 
56 
 
 
 
 
 
 
 
 
 
57 
 
 
 
 
 
 
 
 
 
58 
 
 
 
 
 
 
 
 
 
59 
 
 
 
 
 
 
 
 
 
60 
 
 
 
 
 
 
 
 
 
61 
 
 
 
 
 
 
 
 
 
62 
 
 
 
 
 
 
 
 
 
63 
 
 
 
 
 
 
 
 
 
64 
 
 
 
 
 
 
 
 
 
65 
 
 
 
 
 
 
 
 
 

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Version 3: 18 July 2016 
 
66 
 
 
 
 
 
 
 
 
 
67 
 
 
 
 
 
 
 
 
 
68 
 
 
 
 
 
 
 
 
 
69 
 
 
 
 
 
 
 
 
 
Method Comparison Data Collection Sheet (Case Report Form)   
 
 
 
Page 3 of 3 
 
 
 
System Name 
 
 
 
 
 
 
 
 
System Serial #  
 
 
 
 
 
 
 
Cart Lot # 
 
 
 
 
 
 
 
 
Specimen ID 
Total Chol 
mg/dL or mmol/L 
Total Chol 
mg/dL or 
mmol/L 
Reference 
Analyzer 
Result Total 
Chol  (mg/dL 
or mmol/L) 
Trigs 
mg/dL or 
mmol/L 
Trigs 
mg/dL/ or 
mmol/L 
Reference 
Analyzer 
Result Trig 
(mg/dL or 
mmol/L) 
HDL Chol 
mg/dL or 
mmol/L 
HDL Chol 
mg/dL or 
mmol/L 
Reference 
Analyzer 
Result HDL 
Chol(mg/dL or 
mmol/L) 
70 
 
 
 
 
 
 
 
 
 
71 
 
 
 
 
 
 
 
 
 
72 
 
 
 
 
 
 
 
 
 
73 
 
 
 
 
 
 
 
 
 
74 
 
 
 
 
 
 
 
 
 
75 
 
 
 
 
 
 
 
 
 
76 
 
 
 
 
 
 
 
 
 
77 
 
 
 
 
 
 
 
 
 
78 
 
 
 
 
 
 
 
 
 
79 
 
 
 
 
 
 
 
 
 
80 
 
 
 
 
 
 
 
 
 
81 
 
 
 
 
 
 
 
 
 
82 
 
 
 
 
 
 
 
 
 
83 
 
 
 
 
 
 
 
 
 
84 
 
 
 
 
 
 
 
 
 
85 
 
 
 
 
 
 
 
 
 
86 
 
 
 
 
 
 
 
 
 
87 
 
 
 
 
 
 
 
 
 
88 
 
 
 
 
 
 
 
 
 
89 
 
 
 
 
 
 
 
 
 
90 
 
 
 
 
 
 
 
 
 
91 
 
 
 
 
 
 
 
 
 
92 
 
 
 
 
 
 
 
 
 
93 
 
 
 
 
 
 
 
 
 
94 
 
 
 
 
 
 
 
 
 
95 
 
 
 
 
 
 
 
 
 
96 
 
 
 
 
 
 
 
 
 
97 
 
 
 
 
 
 
 
 
 
98 
 
 
 
 
 
 
 
 
 
99 
 
 
 
 
 
 
 
 
 
100 
 
 
 
 
 
 
 
 
 
 
Name of Person Collecting Data: _______________________________ 
Signature of Person Collecting Data: ____________________________ 
Date: ________________________ 
 
 

75 
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3. Total Cholesterol, Triglyceride and HDL Cholesterol 
Summary Report Template 
 
 
 
 
 
 
 

76 
Version 3: 18 July 2016 
 
 
 
Total Cholesterol, Triglyceride and HDL Cholesterol Summary Report  
Date:  
Report Author:  
Study Site:  
Study Site Principal Investigator/Department:  
Materials and Methods 
See Protocols I-IV. Any deviations from protocol should be noted here.  
Summary of Results 
I. Precision Studies 
Table 1. Total Cholesterol Within-Day Precision for Allegro 
 
 
Low 
Cholesterol 
Mid 
Cholesterol 
High 
Cholesterol 
Mean mg/dL or mmol/L 
 
 
 
Standard Deviation 
 
 
 
%CV 
 
 
 
N 
 
 
 
Matrix 
 
 
 
 
 
Table 2. Triglyceride Within-Day Precision for Allegro 
 
 
Low 
Triglyceride 
Mid 
Triglyceride 
High 
Triglyceride 
Mean mg/dL or mmol/L 
 
 
 
Standard Deviation 
 
 
 
%CV 
 
 
 
N 
 
 
 
Matrix 
 
 
 
 
 
 

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Table 3. HDL Cholesterol Within-Day Precision for Allegro 
 
 
Low 
Cholesterol 
Mid 
Cholesterol 
High 
Cholesterol 
Mean (mg/dL) 
 
 
 
Standard Deviation 
 
 
 
%CV 
 
 
 
N 
 
 
 
Matrix 
 
 
 
 
Table 4. Total Cholesterol Between-Day Precision for Allegro 
 
 
Low 
Cholesterol 
Mid 
Cholesterol 
High 
Cholesterol 
Mean (mg/dL) 
 
 
 
Standard Deviation 
 
 
 
%CV 
 
 
 
N 
 
 
 
Matrix 
 
 
 
 
Table 5. Triglyceride Between-Day Precision for Allegro 
 
 
Low 
Triglyceride 
Mid 
Triglyceride 
High 
Triglyceride 
Mean (mg/dL) 
 
 
 
Standard Deviation 
 
 
 
%CV 
 
 
 
N 
 
 
 
Matrix 
 
 
 
 
Table 6. HDL Cholesterol Between-Day Precision for Allegro 
 
 
Low HDL 
Chol 
Mid HDL 
Chol 
High HDL 
Chol 
Mean (mg/dL) 
 
 
 
Standard Deviation 
 
 
 
%CV 
 
 
 
N 
 
 
 
Matrix 
 
 
 
 
 

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Version 3: 18 July 2016 
 
 
III. Method Comparison  
Table 11. Summary of Linear Regression Analysis Results: Total Cholesterol, Triglycerides and HDL Cholesterol 
for Allegro 
 
 
Total Cholesterol 
Triglyceride 
HDL Cholesterol 
Slope 
 
 
 
y-intercept (mg/dL) 
 
 
 
R2 
 
 
 
N 
 
 
 
Total Cholesterol Range  
mg/dL or mmol/L 
 
 
 
Triglyceride Range 
mg/dL or mmol/L 
 
 
 
HDL Cholesterol  
mg/dL or mmol/L 
 
 
 
 
 
 
 
 
Table 12. Summary of Bland-Altman Results 
 
Total Cholesterol 
Triglyceride 
HDL Cholesterol 
Mean Bias (mg/dL) 
 
 
 
95% Limits of Agreement 
 
 
 
         Lower 
 
 
 
         Upper 
 
 
 
Total Cholesterol Range  
mg/dL or mmol/L 
 
 
 
Triglyceride Range 
mg/dL or mmol/L 
 
 
 
HDL Cholesterol  
mg/dL or mmol/L 
 
 
 
 
 
 
 

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Summary Findings 
Allegro demonstrated …. 
 
 
This report has been reviewed and approved by: 
 
Signature of Study Site Principal Investigator                                                                               Date 
 
 
 
Signature of MASA Director                                                                                                              Date 
 
 
 
 

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