User Performance of the L-PLUS Blood Glucose Monitoring System
Conditions: Diabetes Mellitus
Sponsor: Ascensia Diabetes Care

Clinical Trial Protocol GCA-PRO-2019-002-01 vs 10June2019  
Ascensia Diabetes Care 
Project:00096,   Author: MT Viggiani 
 
CONFIDENTIAL 
 
Page 1 of 34 
 
User Performance of the L-PLUS Blood Glucose Monitoring System 
Protocol GCA-PRO-2019-002-01 
Sponsor: 
Ascensia Diabetes Care   
Global Clinical Affairs 
100 Summit Lake Drive 
Valhalla, NY  10595 
 
NCT number: 04033315 
 
 
  
 
 
 
 
Pragathi Shelat, B.Pharm, MS 
Clinical Research Scientist 
Global Clinical Affairs 
 
 Date 
 
Maria Teresa Viggiani, MS, 
CCRA 
Clinical Trial Manager 
Global Clinical Affairs 
 
 
 
Date 
Scott Pardo, PhD, PStat® 
Deputy Director,  Statistics and Data 
Management 
Global Clinical Affairs 
 Date 
 
Julie Martel, PhD 
Global Head, 
Clinical Development and 
Operations 
 
 
Date 
 
  
 
 
 
 
James Venosky, NPDP 
Program Manager - LCM,   
L-PLUS Project 
 
 
 
 
 
 Date 
 
Colleen Burdel 
Regulatory Affairs Manager,  
Global Regulatory Affairs 
 
Date 
 
 

Clinical Trial Protocol GCA-PRO-2019-002-01 vs 10June2019  
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I have read this protocol and agree to conduct the study as outlined.  I also agree that prior to seeking 
approval from an Institutional Review Board (IRB), the Ascensia Diabetes Care study manager must 
approve any changes to the protocol. 
 
SITE:  Rainier Clinical Research Center, Inc 
 
 
Principal 
Investigator: 
 
Leslie J. Klaff, MD 
Rainier Clinical Research Center, Inc 
723 SW 10th Street, Suite 100 
Renton, WA 98057 
Phone: (425) 251-1720 
Fax: (425) 251-1723 
Email: ljklaff@rainier-research.com 
 
 
__________________  _______________________     _____________ 
             Signature 
Printed Name 
       Date 
 
 
 
This protocol and the data obtained from the study are confidential and may not be disclosed without prior 
written consent of Ascensia Diabetes Care. 

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Table of Contents 
 
1.0 BACKGROUND INFORMATION 
 
          5 
2.0 STUDY OBJECTIVES 
 
          5 
3.0 STUDY DESIGN SUMMARY 
 
          6 
4.0 INCLUSION/EXCLUSION CRITERIA AND SUBJECT RECRUITMENT  
          7 
5.0 MATERIALS AND METHODS 
 
          9 
6.0 RISK / BENEFIT ASSESSMENT 
 
        20 
7.0 ADVERSE EVENT MANAGEMENT AND MEDICAL DEVICE REPORTING         21 
8.0 STUDY TERMINATION AND SUBJECT WITHDRAWAL CRITERIA 
 
        23 
9.0 REGULATORY 
 
        24 
10.0 DATA COLLECTION AND MANAGEMENT 
 
        26 
11.0 DATA ANALYSIS 
 
        26 
12.0 ADMINISTRATION 
 
        27 
13.0 SPONSOR REGULATORY BINDER 
 
        28 
 
APPENDIX A - INSTR. FOR CLEANING AND DISINFECTING METERS 
 
        29 
APPENDIX B - INSTR. FOR COLLECTION/PROCESSING BLOOD FOR YSI 
        30 
APPENDIX C - INSTR. FOR HCT SAMPLE PREPARATION & TESTING 
 
        32 
APPENDIX D - HYPOGLYCEMIA PROTOCOL AT RAINIER 
 
        33 
ATTACH. 1:   PROPER HANDLING OF ASCENSIA YSI SERUM CONTROLS 
        34 
 
 
 
 
 
 

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List of Abbreviations 
 
 
AE 
Adverse Event 
AST 
Alternative site test 
BG 
Blood Glucose 
BGM 
Blood Glucose Meter 
BGMS 
Blood Glucose Monitoring System 
CLSI 
Clinical and Laboratory Standards Institute  
CSR 
Clinical Study Report 
EDC 
Electronic Data Capture 
ICF 
Informed Consent Form 
IRB 
Institutional Review Board 
ISO 
15197:2013 
(and EN ISO 
15197:2015) 
In vitro diagnostic test systems – Requirements for blood–glucose 
monitoring systems for self-testing in managing diabetes mellitus. 
Issued 15May2013, International Organization for Standardization, 
Switzerland. 
Note: All references to ISO:15197:2013 in this protocol also encompass 
EN ISO 15197:2015. 
L-PLUS               
Lightning PLUS 
PI 
Principal Investigator 
PWD 
People with diabetes 
QRG 
Quick Reference Guide 
UADE 
Unanticipated Adverse Device Effect 
UG 
User Guide 
YSI 
YSI Glucose Analyzer™ (YSI Life Sciences, Yellow Springs, OH) 
 
 
 

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1.0 Background Information 
L-PLUS (Lightning PLUS) is the code name of a blood glucose monitoring system (BGMS) 
consisting of a new meter and marketed CONTOUR PLUS® test strip.  The proposed brand name 
for this BGMS is “Contour Plus ELITE”.  This system will be marketed outside the US.  The meter 
can wirelessly transmit BG results using Bluetooth® Low Energy Technology to the meter mobile 
application that will be available on specified versions of the iOS and Android mobile platforms.   
This clinical trial will assess the performance (accuracy) of the L-PLUS meter by lay users enrolled 
as subjects in the study, and by health care professionals (also called study staff).   
The trial will follow the requirements and procedures described in ISO 15197:2013 International 
Standard.  Results will be evaluated and reported for the L-PLUS system according to the ISO 
15197 standard with one report.   
2.0  Study Objectives    
Study objectives 2.1 and 2.2.1 to 2.2.3 below are in accordance with ISO 15197:2013, Section 8. 
2.1   Primary Objectives, ISO 15197:2013:    
2.1.1 Fingerstick Test: Obtain L-PLUS BGMS performance data in the hands of lay users with 
diabetes using fingerstick capillary blood obtained with Microlet® NEXT lancing device. 
• 95% of glucose results shall fall within ±15mg/dL of laboratory method at glucose 
concentration <100 mg/dL and within ±15% at glucose concentrations ≥100 
mg/dL 
2.2 Secondary Objectives, ISO 15197:2013: Failure of any sub-objectives listed below does not 
constitute the failure of the secondary objective. 
2.2.1 Alternative Site Test (AST): Obtain L-PLUS BGMS performance data in the hands of lay 
users with diabetes using capillary blood from the palm site obtained using Microlet NEXT 
lancing device with the AST endcap.  Acceptance criteria will be the same as fingerstick 
2.1.1.  
2.2.2 Venous Blood Test: Obtain L-PLUS BGMS performance data using venous blood from 
lay users with diabetes.  Acceptance criteria will be the same as fingerstick 2.1.1. 
2.2.3 Subject Questionnaire 1:  The questionnaire will include statements about the L-PLUS 
BGMS instructions for use (product labeling) and ease of use of the basic blood testing 
operations of the meter system.  These statements will be given a rating by the subjects 
with diabetes (1=strongly disagree, 2=disagree, 3=neutral, 4=agree, 5=strongly agree), and 
frequency distributions will be tabulated.  Results will be compared to the following 
criteria:  
• 
Acceptance criteria: There must be at least a 90% probability that a user would 
respond with a score of 3 or higher to statements concerning instructions for use 
or ease of use.** 
 
**This is a “total population” (hypothesis test) requirement for the pool of all subjects with diabetes. 

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Study objectives 2.2.4 to 2.2.6 below are internal objectives. 
2.2.4 Fingerstick Test :  Obtain L-PLUS BGMS performance data in the hands of lay users with 
diabetes using fingerstick capillary blood obtained with Microlet NEXT lancing device. 
• Test hypothesis that there is at least a 95% probability that meter results would 
be within ±10.5mg/dL of laboratory method at glucose concentrations <100 mg/dL 
and within ±10.5% at glucose concentrations ≥100 mg/dL** 
2.2.5 Study staff test:  Obtain L-PLUS BGMS performance data in the hands of trained study 
staff testing subjects’ fingerstick blood obtained with Microlet NEXT lancing device. 
• Test hypothesis that there is at least a 95% probability that meter results would 
be within ±15mg/dL of laboratory method at glucose concentrations <100 mg/dL 
and within ±15% at glucose concentrations ≥100 mg/dL** 
2.2.6 Subject Questionnaire 2 :  This questionnaire for subjects with diabetes will include 
statements about user preferences and feedback regarding the features of the L-PLUS 
BGMS in routine diabetes management.  These statements will be given a rating by the 
subjects (1=strongly disagree, 2=disagree, 3=neutral, 4=agree, 5=strongly agree), and 
frequency distributions will be tabulated.  
• Acceptance criteria: There are no pass/fail criteria for Questionnaire 2.   
 
 
3.0 Study Design Summary 
This study is designed to satisfy the ISO 15197:2013, Section 8  requirements. The ISO 
15197:2013 standard states that the study population shall be people with diabetes only. 
The study will be conducted at a minimum of one clinical site and enroll a total of approximately 
130 lay users.    
For this study, there must be at least 120 evaluable results each for fingerstick and AST self-test 
results, as well as for staff testing subject fingertip and venous blood using the L-PLUS BGMS. 
Subject enrollment will continue until the required evaluable samples are collected.   
Sites are not required to replace subjects who did not have evaluable venous blood results, once 
120 venous tests are completed. 
The subjects will be 18 years and older.  Each subject will make one visit, lasting approximately 
1 hour at the clinical site.  Study sessions will be conducted by study staff members who have been 
trained on all aspects of the protocol, the use of the L-PLUS  system, data collection, and GCP.   
All subjects will complete the informed consent process before performing any study procedures.   
Procedures - All Subjects  
Subjects will be assigned the L-PLUS  meter and randomized to use one of three Contour Plus test 
strip lots (Red, Green, or Blue) throughout the visit.   
 

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The untrained subjects, who have never used Ascensia’s Contour Next (Lightning Next) and 
Lightning PLUS (L-PLUS or Contour Plus ELITE) BGMS previously , will be sequestered in such 
a way that they cannot observe or be influenced by the testing technique of other study participants.  
They will learn to use the L-PLUS meter by reading the corresponding User Guide (UG) and Quick 
Reference Guide (QRG).  Each subject will perform one fingerstick self-test using a Microlet 
NEXT or similar lancing device and the L-PLUS meter.  Immediately following the subject self-
test, a Tenderlett™ or similar device will be used by the study staff on the subject’s finger 
(preferably the same hand) to collect a capillary sample for testing on the YSI Glucose Analyzer.   
Immediately following the Tenderlett lancing, subjects will lance their palm using the Microlet 
NEXT lancing device (with AST clear endcap) and will perform a self-test using the meter.   
Immediately following the AST test, the staff person will perform a fingerstick test on the subject 
with the Microlet device and the L-Plus meter.  
All subject capillary and AST self-test results and study staff fingerstick results will be compared 
to YSI results from blood obtained from the Tenderlett fingerstick. 
Hematocrit will be measured for all subjects.    
All Subjects will then complete a questionnaire (Questionnaire 1) to provide feedback on the meter 
system and instructions for use (UG and QRG).   
Subjects will be given a venipuncture.  The venous blood will be tested by the study staff using 
the L-PLUS  system and the YSI lab analyzer.  The venipuncture may be performed any time after 
the subject is enrolled (i.e. completed informed consent and passed inclusion/exclusion criteria). 
After all self-testing and Questionnaire 1 is completed, staff will demonstrate to the subjects 
several features of the meter system that would not necessarily be experienced during the brief 
accuracy testing.  The subjects will be given a second questionnaire (Questionnaire 2) to provide 
feedback about the new features of the system.   
 
4.0 Inclusion/Exclusion Criteria and Subject Recruitment 
4.1 
 Inclusion Criteria: 
4.1.1 Males and females, 18 years of age and older 
4.1.2 People with diabetes (PWD) – type 1 or type 2 
4.1.3 Ability to speak, read and understand English.  Subjects must demonstrate ability to read a 
paragraph from the first page of the UG to qualify for the study. 
4.1.4 Willing to complete all study procedures 
4.2 
Exclusion Criteria: 
4.2.1 Hemophilia or any other bleeding disorder 
4.2.2 Pregnancy (self-reported) 
4.2.3 Physical, visual or neurological impairments that would make the person unable to perform 
testing with the BGMS.  

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4.2.4 Previous participation in a blood glucose monitoring study using the Ascensia’s Contour 
Next (Lightning Next) and Lightning PLUS (L-PLUS or Contour Plus ELITE) BGMS. 
4.2.5 Working for a medical laboratory, hospital or other clinical setting that involves training on 
or clinical use of blood glucose monitors. 
4.2.6 Being in this trial during or less than two days since  xylose absorption testing (Xylose in the 
blood is known to cause interference).  However, subjects can be enrolled as soon as 2 days 
after the test being performed (25gms of oral xylose administered during this test is cleared 
within 480 mins or 8).  
4.2.7 Working for a competitive medical device company, or having an immediate family member 
or someone who is not a family member but is living within the household of someone who 
works for such a company.   
• Immediate family members are the subject’s parents, spouse, children, and siblings, 
including the parent’s spouse; step-children and adopted children and their spouses.   
• A competitive medical device company is a company that provides a medical device 
or components of a device that is related to diabetes.   
For example, people who are not eligible are those who work for companies that create or 
manufacture the following (or a company that is in a partnership with a company that 
provides such devices): lancing devices, blood glucose monitoring systems, continuous 
glucose monitoring systems, insulin pens, or systems related to the measurement of HbA1c.   
People who are eligible are those who work for companies associated with products such as 
wound dressings, medications or dietary products. 
4.2.8 A condition, which in the opinion of the investigator or designee, would put the person or 
study conduct at risk.  The reason for exclusion will be clearly documented by investigator 
or designee on the subject disposition form. 
 
4.3 
Subject Recruitment 
4.3.1 The following population criteria are study goals, with the site recruiting to meet these goals: 
• At least 130 subjects who are PWD (type 1 or type 2), and who have never previously 
used Ascensia’s Contour Next (Lightning Next) and Lightning PLUS (L-PLUS or 
Contour Plus ELITE) BGMS, will be enrolled.  This is to have a total of at least 120 
evaluable results each for fingerstick and AST self-test results, as well as staff testing 
subject fingertip and venous blood using the L-PLUS BGMS. 
• At least 70% of subjects will be younger than age 65.  
• At least 20% of subjects will have type 1 diabetes.  
4.3.2 No laboratory tests (e.g., pregnancy) are required to assure qualification.  Subjects’ verbal 
responses will be accepted and recorded on  source documents. 
 
 

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5.0 Materials and Methods 
5.1 
Resources Supplied by Investigational Site 
5.1.1 Staffing 
1. Principal Investigator and, as needed, Sub-Investigator(s) 
2. Study Coordinator 
3. Two or more study staff to conduct the study  
4. Staff person(s) to measure hematocrit, prepare samples for YSI analysis, and perform YSI 
analysis of venous and capillary blood (plasma) and serum controls  
5. Data entry person for EDC data entry (Note: This requires EDC training.) 
5.1.2 Other 
1. Final informed consent forms (ICF), and other necessary supporting documents, as 
appropriate 
2. Computer with internet connection for site and monitor access to EDC system 
3. Facility with adequate space for conducting all testing.  Site facilities for conducting 
subject visits must allow sufficient privacy so subjects cannot observe other subjects’ 
BGMS testing procedures.  Testing area must also be conducive to allow subjects to focus 
on testing (i.e. background distraction to be kept to a minimum). 
4. YSI 2300 analyzers, reagents, and supplies. Up to date maintenance documentation will 
be required.  
5. Supplies to treat any hypoglycemic events 
6. Venipuncture supplies 
7. Laboratory supplies including: disposable pipets, gloves, alcohol wipes, band aids, gauze, 
parafilm, disposable benchtop covers   
8. Tenderlett™ or similar single-use lancing device 
9. Micro collection tubes with lithium heparin anticoagulant (no gel separator) 
10. Hand warmers 
11. Biohazardous bags and sharps containers  
12. Microhematocrit tubes without anticoagulant (for testing venous blood) 
13. Microhematocrit tubes with anticoagulant (for testing capillary blood) 
 
5.2 
Resources Supplied by Ascensia 
5.2.1 Staffing 
1. Ascensia Study Manager 
2. Study Monitor(s)  
 

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5.2.2 Materials 
1. Protocol, EDC system, paper forms (CRF) and questionnaires, as needed 
2. ICF template 
3. Investigational L-PLUS meters (Note: Meters will be provided with Bluetooth turned ON 
and will have it ON throughout the study. Note: The meters are labeled with the proposed 
brand name only – Contour Plus ELITE.) 
4. Investigational L-PLUS User Guides (UG) and Quick Reference Guides (QRG). Note: 
UG and QRG refer to the proposed meter brand name only – Contour Plus ELITE.)  
5. CONTOUR PLUS test strips - 3 lots labeled as RED, GREEN, BLUE (R, G, B) and 
Contour Plus Test Strips Inserts.  A randomization table for the assignment of strip lot 
will be provided to the site.  Note: CONTOUR PLUS test strips are not registered for sale 
in the US. 
6. Contour PLUS control solution (Normal level equivalent to Level 2) and Contour Plus 
control solution inserts.  Note: CONTOUR PLUS control solution is not registered for 
sale in the US. 
7. Microlet NEXT lancing device systems including clear, AST endcap.  (All subjects will 
be provided new lancing devices and lancets; lancing devices will not be shared among 
subjects.) 
8. Thermometer / hygrometer  
9. Statspin™ microcentrifuge and tube rotors  
10. Microhematocrit tube reader (or equivalent hematocrit measurement system), hematocrit 
rotors, sealant 
11. Ascensia serum glucose traceability controls (four levels) 
12. Disinfectant wipes for meters (listed in Appendix A) 
13. Package delivery forms and envelopes (UPS or Fed Ex) 
 
5.3 
Site Staff Pre-Study Orientation 
The Ascensia Study Manager, or designee, will review all study documents, procedures (including 
procedures for data collection and management), GCP, and usage of the L-PLUS BGMS with the 
site staff during the Site Initiation Visit.  All site staff involved in subject testing sessions will 
successfully perform at least one control solution test with each L-PLUS BGMS.  Study-specific 
EDC training will also be provided to study staff as necessary and training will be documented by 
the site.  
 
5.4 
Safety and Disinfection Procedures: 
1. Universal precautions will be used throughout the study. 
2. Only study staff will perform disinfection procedures per Appendix A.  Subjects will not 
perform disinfection.  

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3. It is intended that a new L-PLUS meter will be provided to each subject.  At the end of the 
subject’s visit, the meters will be disinfected and stored until they are returned to Ascensia. 
4. A new Microlet NEXT or similar lancing device will be provided to each subject.  At the end 
of the subject’s visit, the lancing device will either be discarded or offered to the subject to 
take home. 
5. The lancets used in the study (Tenderlett and Microlet lancets) will be disposed of in a 
biohazard sharps container after use.  
6. Microlet lancets must be changed for each lancing by the subject.  (Insert, remove, and 
discard into sharps container.)   
7. Disposable exam gloves will be worn by site staff during study procedures where the risk of 
transmission of disease is present.  These procedures include performing blood glucose 
measurements, sample processing, and laboratory procedures involving blood samples.  A 
new pair of gloves will be used when working with each subject and will be changed between 
subjects. 
8. All test strips are single-use and will be appropriately discarded after use. 
9. Disposable bench top covers should cover the work area and must be changed between 
subjects. 
10. A marketed BGM device may be used for determining the subject’s glucose level during the 
study as needed for safety reasons or to ascertain the glucose level for study reasons.  The 
meter will be disinfected after use and store away for return. 
5.5 
Study Staff Quality Control Testing of Meters  
Prior to enrollment of the first subject, the study staff will perform testing with Contour Plus 
normal level control solutions on all L-PLUS  meters.  All staff control testing will be done out of 
sight of the study subjects.  If the results are out of range, troubleshooting will be performed (see 
meter UGs) and the test will be repeated.  If the meter tests are out of range after troubleshooting 
is complete, the meter will be removed from the study.  This will be documented on the Problem 
Reporter form, as described in Section 5.15 ‘Device Problem Reporting’.  
5.6 
Subject Enrollment 
5.6.1 The study staff will perform the following when the subject arrives at the clinical trial site: 
1. Complete the informed consent process.  
2. Assign a unique number to each subject. 
5.6.2 The study staff will document subject demographics (including both screen-failed and 
enrolled subjects), diabetes history, medical conditions and medications information. 
Subjects will also be asked when they last ate, took diabetes medications, and performed 
vigorous exercise.  Vigorous exercise is considered to be more than the subject’s personal 
normal level. 
5.6.3 According to the randomization schedule, subjects will be assigned to use one of three 
Contour Plus test strip lots (R,B or G) throughout the visit.  

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5.6.4  After enrollment, subject venipuncture may be performed at any time during the visit. 
 
5.7 
Tests (See Table 5.1 for test scheme.) 
Each subject will be provided with all the materials needed for self-testing during the visit: one L-
PLUS meter, a Microlet NEXT or similar lancing device, lancets, Contour PLUS test strips, 
associated meter UGs and QRGs.  The subject will be given time to review the UG and QRG to 
learn how to use the system but no training will be provided.   
The subject must wash his/her hands thoroughly with warm water and soap, rinse and dry well.   
5.7.1 Fingerstick Testing  
1. The subject will perform a fingerstick using the Microlet NEXT lancing device and test 
his/her blood on L-PLUS meter.  The study staff person will record the time and test result 
of the successful meter test on the appropriate form.  For instructions on handling error codes 
and repeat testing, refer to protocol section 5.10 ‘Errors’. 
2. Immediately, but at least within 5 minutes of the fingerstick self-test, the study staff person 
will lance the subject’s finger (preferably the same hand as above) with a Tenderlett device 
in order to collect enough blood for testing on the YSI Analyzer (approx. 200–300 uL).  The 
time of this Tenderlett lancing will be recorded. 
3. If adequate quantity of blood is not obtained from the Tenderlett lancing, 2 additional 
Tenderlett lancings are allowed (total of up to 3 Tenderlett lancings).  The blood is to be 
combined into one tube for YSI analysis.  
4. The blood sample will be centrifuged as soon as possible after sample collection but not 
longer than 15 minutes after the meter test.  The study staff person will record the time of 
centrifugation.  
5. In order to reduce glycolysis of the plasma, it is recommended that the plasma be transferred 
from the microtainer tube to a micro/dispo tube or similar container, prior to analysis on the 
YSI Analyzer. 
6. The plasma sample will be assayed on the YSI Analyzer: 1 sip = 1 result from black probe 
and 1 result from white probe.  See 5.12.3  for details on testing of plasma samples. 
7. Immediately following the subject plasma sample, a serum control close to the level of the 
subject plasma will be tested. (No re-calibration is required).  The serum control test result 
should be within the range shown in Table 5.2.  See 5.12.3  for details. 
8. Additional details regarding processing of blood samples for the YSI Analyzer are found in 
Appendix B.  
9. If the time from meter test to blood centrifugation exceeds 15 minutes,  the test is considered 
non-evaluable.  This will not be considered a protocol deviation.  Times of meter tests and 
centrifugation will be recorded.  

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5.7.2  AST (palm) testing 
1. Only proceed to AST testing if an adequate blood sample is obtained from Tenderlett lancing 
for YSI analysis. 
2. The same meter and test strip lot used for fingerstick testing will be used for AST testing. 
3. Immediately following Step 5.7.1, the subject will perform an AST (palm) stick using the 
lancing device (new lancet), preferably on the same hand as the fingerstick lancing, and test 
his/her blood on the meter.  
4. If adequate blood is not obtained from the Microlet palm lancings for the meter test, up to 2 
additional Microlet palm lancings are allowed (total of up to 3 AST palm lancings). 
5. The time of the successful AST meter test will be recorded.  The time interval between the 
blood centrifugation and the AST meter test should be as soon as possible, and within 15 
minutes or less.  
6. If the time from meter test to blood centrifugation exceeds 15 minutes, the test is considered 
non-evaluable.  This will not be considered a protocol deviation.  Times of meter tests and 
centrifugation will be recorded. 
5.7.3 Study Staff test 
1. Immediately following the AST testing, the study staff person will perform a fingerstick 
lancing using the same Microlet lancing device (new lancet) on the subject, preferably on the 
same hand as the other fingerstick lancing. 
2. The staff personnel will test the fingerstick blood using the same meter and test strip lot used 
by the subject.   
3. The study staff will record the time of the meter test.  The time interval between the blood 
centrifugation and the final staff meter test should be as soon as possible, and within 15 
minutes or less.    
4. If the time from meter test to blood centrifugation exceeds 15 minutes, the test is considered 
non-evaluable.  This will not be considered a protocol deviation.  Times of meter tests and 
centrifugation will be recorded. 
5.8 
Venous Blood Testing by Study Staff   
5.8.1 The venipuncture and subsequent test may be performed at any time during the subject visit.   
5.8.2 A venipuncture will be performed by a trained site phlebotomist.  The subject’s blood will 
be collected into one 4 mL heparin-coated Vacutainer tube.   
5.8.3 The study staff person will place one drop of venous blood onto Parafilm®, then immediately 
perform one test with the same lot of test strips and the same meter used for capillary tests.  
Record the time of the successful venous meter test. (Refer to Section 5.10 for information 
on handling error codes and repeat testing.) 
5.8.4 The venous blood will be centrifuged as soon as possible after the meter test (within 15 
minutes or less).  Record the centrifugation time. If the time from meter test to blood 
centrifugation exceeds 15 minutes, the test is considered non-evaluable.  This will not be 
considered a protocol deviation.  Times of meter tests and centrifugation will be recorded. 

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5.8.5 The venous plasma sample will be assayed on the YSI Analyzer: 1 sip = 1 result from black 
probe and 1 result from white probe.  See 5.12.3 for details on testing of plasma samples and 
Appendix B regarding processing blood samples for the YSI analyzer. 
5.8.6 Up to three venipuncture attempts are allowed per subject, if subject and study staff agree, 
in order to obtain sufficient venous blood.  If the venipuncture is ultimately unsuccessful, 
this will be recorded and will not be considered a protocol deviation.  
 
Table 5.1 – Subject Testing Schematic 
Step 
Test type 
Operator 
Description 
1 
Fingerstick 
Self-test 
Subject 
 Subject lances finger – Microlet NEXT lancing device.  
 Test meter with 1 lot of test strip  (Repeat 2 more times, only if needed). 
 Record time of successful meter test. 
2 
Tenderlett 
deep stick 
Study 
staff 
 Immediately following Step 1 meter  test (≤  5 min), staff lances subject 
fingertip (up to 3 lancings allowed). 
 Record time of lancing.  
 Collect blood in Li Heparin tube (approx. 200 – 300 uL).  
 Centrifuge blood and record time (should be within 15 min of meter test in step 
1). 
3 
AST palm 
Self-test 
Subject 
 Immediately following Step 2, subject lances palm – Microlet NEXT lancing 
device. 
 Test with same test strip lot and meter as above.  (Repeat 2 more times, only if 
needed.) 
 Record time of successful meter test (should be within 15 min of centrifugation 
in Step 2). 
4 
Study staff 
test 
Study 
staff 
 Immediately following Step 3, staff lances subject fingertip - Microlet NEXT 
lancing device. 
 Test with same test strip lot and meter as above.  (Repeat 2 more times, only if 
needed.) 
 Record time of successful meter test (should be within 15 min of centrifugation 
in Step 2). 
5 
Venous test 
Study 
staff 
 May be done prior to Steps 1-4. 
 Staff may perform up to 3 venipuncture attempts. 
 Collect blood in 4 ml Vacutainer tube with heparin anticoagulant. 
 The next two sub-steps may be performed in either order; they should be within 
15 minutes maximum of each other.  
o 
Staff  tests blood  (out of view of subject) with same meter and strip lot as 
fingerstick test. 
o 
Centrifuge blood and record time (should be within 15 min of venous 
blood meter test). 
6 
Hematocrit 
Study 
staff  
 Fill 2 microhematocrit tubes  (only 1 tube to be measured).  
 Blood is from capillary or venous blood. 
7 
Question-
naire 1 
Study 
staff 
 Study staff administers Questionnaire 1 to all subjects. 
8 
Question-
naire 2  
Study 
staff 
 Study staff demonstrates BGMS features to subjects.  
 Study staff administers Questionnaire 2 to subjects. 
 

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5.9 
General 
5.9.1 If adequate blood is not obtained from the subject or staff Microlet lancings, up to 2 
additional lancings are allowed in each case (up to a total of 6 fingertip Microlet lancings). 
5.9.2 If adequate blood is not obtained from the staff Tenderlett lancing, up to 2 additional lancings 
are allowed in each case (up to a total of 3 fingertip Tenderlett lancings). The blood from 
each lancing should be combined into one Microtainer tube.  
5.9.3 If adequate blood is not obtained from the Microlet AST palm lancings, up to 2 additional 
palm lancings are allowed (total of up to 3 AST palm Microlet lancings). 
5.9.4 If adequate blood is not obtained from the first venipuncture attempt, up to 2 additional 
attempts are allowed if subject and study staff agree (total of up to 3 venipuncture attempts). 
5.9.5 If the time from meter test to blood centrifugation exceeds 15 minutes, the test is 
considered non-evaluable.  This will not be considered a protocol deviation.  Times of meter 
tests and centrifugation will be recorded.  
5.9.6  Hand warmers may be used throughout the testing to increase blood flow. 
5.9.7  It is preferred that all fingerstick and AST lancings are from the same hand if possible, 
throughout the study 
5.9.8 Study staff will record all meter results on the appropriate form.  If meters have turned off, 
meter memory will be reviewed and the results will be obtained from meter memory. 
5.9.9 Study staff will record in the comments section of the meter testing form, any obvious testing 
errors by the subject or staff as appropriate for staff tests, and any abnormalities observed 
in the testing procedure or BGMS. 
5.9.10 Note that once the fingerstick testing is started in 5.7.1, the subjects will be advised not eat, 
or use medications until the study staff testing is completed in 5.7.3, to avoid changing 
glucose levels.  The subject can eat/take medicines at any time to avoid or treat 
hypoglycemia, hyperglycemia or an adverse event.  If the subject eats or takes medications, 
the subject’s participation in the study will be discontinued.  
5.10 Errors  
5.10.1 If the meter reports an error code, the instructions shown in the UG should be followed.  
Re-testing is recommended (per the UG).  No more than a total of 3 attempts are allowed. 
5.10.2 If the subject feels that he/she made an error while testing, without any prompting from the 
study staff, he/she will repeat the test.  The reason for the repeated test will be documented 
in the comments section of the form.  No more than a total of 3 attempts are allowed. 
5.10.3 If the study staff feels that he/she made an error while performing staff fingerstick or venous 
testing, he/she will repeat the test.  The reason for the repeated test will be documented in 
the comments section of the form.  No more than a total of 3 attempts are allowed. 
5.10.4 The study staff will record all non-numeric codes - QNS (Quantity not sufficient), NO (Not 
obtainable), and meter error codes as appropriate for the meter tests. 

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5.11   Hematocrit  
Two hematocrit tubes will be filled with either the subject’s venous or capillary blood, and then 
centrifuged.  The hematocrit will be measured from only one tube; the second tube will be collected 
as a backup in case the first tube is not measureable (i.e., tube spun out, broken tube, clay seal 
compromised, etc.).  Subject hematocrit measurement should be within 0-70% in order for the 
subject BG results to be considered evaluable for the ISO 15197 analysis.  
Note:  Microhematocrit tubes with anticoagulant will be used for capillary blood hematocrit 
measurements collected directly off the fingertip and microhematocrit tubes without anticoagulant 
will be used for venous blood. 
Additional details regarding hematocrit measurements are found in Appendix C.  
 
5.12 YSI Glucose Analyzer  
5.12.1 YSI Analyzer Operation and Control Tests 
The YSI Glucose Analyzer will be maintained and operated according to the instructions in the 
manufacturer’s operating manual.   
Before the study begins, the YSI will be set up and appropriate maintenance will be performed.  
The YSI clock will be set to 24 hour format using an accurate time device, such as site central 
clock or atomic clock. 
A set of four glucose control sera1 that have been assayed by a method traceable to one proposed 
for use as a national glucose reference method developed through the combined efforts of the 
Center for Disease Control, the National Bureau of Standards, the American Association for 
Clinical Chemistry, and the FDA will be provided by Ascensia and used to verify the accuracy of 
the YSI Glucose Analyzer2.  These controls will be run for pre-study and daily control tests, and 
will be within the lower and upper limits of the target levels noted in Table 5.2.   The procedure 
for proper thawing and handling of the serum controls is found in Attachment 1.  
Table 5.2: YSI serum control target values and limits, mg/dL  
LOT# 
LEVEL TARGET 
Lower 
Limit 
Upper 
Limit 
7AP204 
2 
48.02 
45.52 
50.52 
7AP304 
3 
95.89 
91.10 
100.69 
7AP404 
4 
193.19 
183.53 
202.85 
7AP505 
5 
388.88 
369.44 
408.32 
 
 
1 Caution:  The glucose traceability controls are human serum-based and must be handled using universal precautions. 
2 Neese J, Duncan P, Bayse D, et al.  Development and evaluation of a hexokinase/glucose-6-phosphate dehydrogenase procedure 
for use as a national glucose reference method. HEW Publication No. (CDC) 77-8330. 1976. Atlanta, Centers for Disease Control. 

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5.12.2 Pre Study and Daily Control Tests  
These four glucose control sera will be used to verify the accuracy of each YSI analyzer before the 
study begins (Pre-study testing).  Six (6) pre-study runs will be performed on each YSI that will 
be used in the trial: 2 runs per day for 3 days.  The Ascensia Study Manager or designee will review 
the YSI pre-study glucose control data before the study begins.  
The daily controls  will be tested: 1 sip = 1 result from white probe and 1 result from black probe.  
Any YSI used on a given day to test subject blood samples will be monitored by assaying the 4 
glucose controls at the beginning of the day.  The YSI will be calibrated before each serum control 
sample is tested.  The sequence of testing will be: 
1. Calibrate the YSI 
2. Test Control 2 (1 sip) 
3. Calibrate 
4. Test Control 3 (1 sip) 
5. Calibrate  
6. Test Control 4 (1 sip) 
7. Calibrate 
8. Test Control 5 (1 sip) 
Site staff will keep a maintenance log for each YSI, including daily operational checks, 
maintenance and membrane changes.  A full set of serum controls should also be tested after YSI 
parts are changed (such as buffers, probes, or membranes).    
5.12.3 Testing of Subject Plasma Samples 
The subject plasma samples (capillary and venous) will be tested using 1 sip per test (=2 results, 1 
from white and 1 from black probe) with calibration of the YSI before the first sip.  A serum 
control (1 sip) will be tested after each subject plasma sample to verify the YSI Analyzer has 
remained in control during the plasma sample test.  The control with target value close to the 
concentration of the subject plasma sample will be tested.  The average of the black and white 
probe results will be verified to be within the ranges shown in Table 5.2.  
If the control result is out of range, the YSI will be recalibrated and the control sample will be 
retested.  If the control result is now within the appropriate range (Table 5.2), the control sample 
is confirmed as appropriate for use.  The YSI analyzer will then be re-calibrated, the plasma will 
be re-tested, and the control will be tested per the protocol’s plasma testing procedure already 
described.  
If the control result is not within the appropriate range after the plasma sample has been tested 
twice, the results for this subject sample will be non-evaluable.  
The sequence of testing for each subject sample will be: 
1. Calibrate the YSI 
2. Test plasma sample (1 sip) 
3. Test control (1 sip) 
4. Verify control result (mean of black and white probe results) is within limits in Table 
5.2.  If it is, stop and record result. 
5. If not, repeat Steps 1, 3 and 4 to verify control sample is satisfactory for use. 
6. When control result is shown to be within range in Table 5.2,  repeat Steps 1-3. 

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If the control is again outside the limits, the results for this subject sample are non-evaluable. 
Glucose values of the YSI replicates (both controls and plasma) should be within 4 mg/dL or 4% 
of each other (i.e. 4 mg/dL for values < 100 mg/dL and 4% for values ≥ 100 mg/dL).  If they are 
not, an additional assay should be run. 
 
5.12.4 Recording YSI Results  
The YSI source documents will be labeled as follows throughout the study to facilitate matching 
glucose results on the source document with results recorded on the CRF: 
Sample identification will include: 
1) Ascensia serum traceability control level or 
2) Subject ID plus either ‘capillary’ or ‘venous’ blood 
The YSI operator or designee will label all samples on YSI tape and record YSI results for all 
plasma and control sample runs on the appropriate source documents. YSI source documents 
(tapes) will be photocopied to preserve the image on the thermal paper and will be stored in 
individual subject’s study file for  source data verification.  Site staff or designees who photocopy 
the YSI tapes will certify (with signature, date, and YSI ID) that the copy is a true and accurate 
representation of the original. 
NOTE:  If glucose results are not to be used due to operator errors, insufficient sample, etc., this 
should be clearly marked on the YSI source document.  The YSI operator will record all non-
numeric codes - QNS (Quantity not sufficient), NO (Not Obtainable), and error codes. 
 
5.13 Subject Questionnaires  
Site staff will administer two questionnaires to each subjects corresponding to L-PLUS  meter.    
5.13.1 Questionnaire 1: Evaluation of Instructions for Use.   
Staff will administer a questionnaire to all subjects who perform fingerstick self-testing after the 
individual meter testing is completed.  The questionnaire will include statements for which a 
numerical score or rating will be provided by subjects (1=strongly disagree, 2=disagree, 3=neutral, 
4=agree, 5=strongly agree).  These numerically scored statements will be tabulated into frequency 
distributions.  The following statements will be subject to the pass/fail criteria listed in the study 
objectives section: 
• I find it easy to do a fingerstick blood test with this meter. 
• The meter display is easy to see and read. 
• It was easy to understand my test results. 
• The instructions (User Guide and Quick Reference Guide) were easy to understand. 
• The instructions clearly explain how to run a test. 
• The instructions clearly explain what to do if an error message is displayed by the 
meter.   Note: an error message will be demonstrated by the study staff if the subject 
does not experience an error message during his/her testing. 

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Per ISO 15197:2013, Section 8.8.2, the questionnaire will also contain a section whereby the 
subject can provide unrestricted comments on their experience about using the BGMS.  This 
questionnaire will be administered by the staff who will record the subject’s responses.  The subject 
will be given an example questionnaire to assist with providing the responses.  
5.13.2 Questionnaire 2: BGMS Features.  
After all testing and the first questionnaire have been completed, the study staff will demonstrate 
to the subjects several features of the BGMS that would not necessarily be experienced during the 
brief accuracy testing.  This will ensure that the subjects are able to understand the features, and 
answer questions in the questionnaire.  
There are no acceptance criteria for this questionnaire.  This questionnaire may be 1) administered 
by the staff who will record the subject’s responses or 2) completed by the subjects and verified 
by the study staff, as per site procedures.  In the first scenario, the subject will be given an example 
questionnaire to assist with providing the responses. 
5.14 Device Problem Reporting 
Any functional problems with the L-PLUS BGMS will be documented by the study staff and 
reported timely to Ascensia.  The study staff should be specific about describing the problem and 
the sequence of events that led to it.  All information will be documented on the Problem Reporter 
Form, including the meter type, serial number and test strip lot.  Malfunctioning BGMs will be 
replaced and this will be documented for study tracking.  If appropriate, such meters will be 
disinfected and shipped back to Ascensia for evaluation. 
5.15 Temperature and Humidity Monitoring  
The study staff will measure the temperature and humidity of the meter and YSI testing area(s) 
twice a day (AM and PM) using an audited thermometer/hygrometer supplied by Ascensia, and 
record the results on the appropriate form.   
5.16  Miscellaneous  
Ascensia representatives may observe subject visits as part of study staff training, study monitoring 
or for troubleshooting problems with the investigational devices.  This will be done under 
supervision of the investigator.  
Ascensia representatives may assist with disinfection of the meters, and perform centrifugation of 
blood samples and measurement of hematocrit.   
Ascensia representatives will not train or interact in any way with the subjects.  The presence of 
Ascensia representatives will be noted in the ICF. 
Subjects will receive nominal compensation for their time and inconvenience. 
 
 

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6.0  Risk / Benefit Assessment 
Risks associated with this study include those associated with obtaining blood from finger, palm 
and venipuncture.   
The following risks are anticipated and will not be reported as Adverse Events (AE). (See section 
7.0 for further details.) 
1) Bleeding: Mild bleeding similar to being stuck with a pin 
2) Swelling (Edema): A mild swelling similar to a mosquito bite 
3) Bruising: Less than one inch across (fingerstick) or less than 2 inches across (venipuncture) 
4) Redness (Erythema): Mild reddening or darkening of the skin in the immediate area around 
the puncture site 
5) Soreness: Mild discomfort at the puncture site resolving (going away) within one to two days 
(fingerstick or palm) or within hours (venipuncture) 
The following risks are unlikely, but possible, and will be reported as anticipated Adverse Events: 
1) Bleeding: Continued bleeding requiring changing a bandage 
2) Bruising: A worsening bruise that is firm to the touch 
3) Redness (Erythema): Bright redness or skin darkening spreading across the area (fingerstick 
or palm) or across the arm (venipuncture) 
4) Soreness: Worsening pain and/or pain that interferes with activity 
5) Infection: Bright redness or skin darkening spreading across the area with warmness and/or 
swelling, and/or red streaking 
6) Nerve injury: Numbness or shooting pain in the forearm 
7) Numbness in the area of the puncture wound 
8) Dizziness 
9) Fainting   
Subjects should be instructed to report such events to the investigator or designee.  
Benefits 
A benefit associated with the study is the sense of well-being gained by contributing to the 
development of improved or new blood glucose monitoring systems, which may be beneficial to 
people with diabetes or other associated diseases or conditions. 
 

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7.0  Adverse Event Management and Medical Device Reporting   
The procedures to be performed under this protocol are considered to be low risk. 
7.1 
Description and Classification 
7.1.1. An AE during a clinical evaluation is “any untoward medical occurrence, unintended 
disease or injury, or untoward clinical signs (including abnormal laboratory findings) in subjects, 
users or other persons, whether or not related to the investigational medical device.”  It is not 
dependent on whether the event is considered to be related to the investigational product or the 
study.  An AE includes events not seen at the beginning of the study, or worsened if present at the 
beginning.  Adverse events can be classified as presented in Table 7.1. 
7.1.2. The next classification to consider is whether or not the AE is Anticipated or Unanticipated.  
In order for an AE to be considered ‘anticipated’ it must be listed in the protocol and ICF prior to 
any occurrence. 
7.1.3. For studies on products that have not yet been cleared for market, it is important that all 
AEs be documented and included in the study file. 
7.1.4. An unanticipated adverse device effect (UADE) is:  Any serious adverse effect on health 
or safety or any life-threatening problem or death caused by, or associated with a device, if that 
effect, problem or death was not previously identified in nature, severity or degree of incidence in 
the investigational plan or application (including a supplementary plan or application), or any other 
unanticipated serious problem associated with a device that relates to the rights, safety or welfare 
of subjects [21CFR812.3(s)]. 
7.1.5. All AEs observed by the Investigator or reported by the subject will be documented on the 
AE form (including all symptoms) and included in the study file.  
7.2 
Events related to hypoglycemia and hyperglycemia 
Hypoglycemia and hyperglycemia can be a common occurrence with subjects who have diabetes.   
If it is found that a subject has a hypoglycemic event, as determined by the Principal Investigator 
(PI) during a study visit, the subject will be treated by the site staff accordingly. Appendices D and 
E note each site’s treatment plan.  
Classification of the hypoglycemic or hyperglycemic event is at the discretion of the PI or 
designee.  If the hypoglycemic or hyperglycemic event is considered by the PI as an AE, it will be 
considered to be an anticipated AE and will be recorded on the AE Form. 
Subjects who are hypo- or hyperglycemic may continue with fingerstick testing if they so choose, 
and if the PI determines that it is safe.  The PI will define what is considered to be a hypoglycemic 
or hyperglycemic state. 
 
 

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Table 7.1: Classification of Adverse Event 
Serious* 
Non-Serious 
An adverse event that leads to:  
a) death;  
b)  a serious deterioration in health of the subject that,  
1) 
resulted in a life-threatening illness or 
injury,  
2) 
resulted in a permanent impairment of a 
body structure or a body function,  
3) 
required in-patient hospitalization or 
prolongation of existing hospitalization,  
4) 
resulted in medical or surgical intervention 
to prevent permanent impairment to body 
structure or a body function;  
c) fetal distress or a congenital abnormality or birth 
defect.  
May be anticipated or unanticipated. 
*EN ISO 14155.  Clinical investigation of medical 
devices for human subjects – Good Clinical Practice 
All adverse events not reported as serious.  At each visit, all 
adverse events observed by the investigator/designate or 
reported by subject must be evaluated and recorded on an 
adverse event form.   
A non-serious adverse event is further classified with respect to 
severity and relationship to the trial product.  A non-serious 
adverse event may be anticipated or unanticipated. 
Further classification with regard to severity 
• 
Mild: Transient symptoms, easily tolerated, no interference 
with subjects daily activities. 
• 
Moderate: 
Marked symptoms, moderate interference 
with subjects daily activities, and tolerable. 
• 
Severe:  Considerable interference with subject’s daily 
activities, not tolerable. 
Relationship to trial product: 
• 
Definitely Related: Causal relationship has been established and documented. 
• 
Probably Related: Good reasons and sufficient documentation to assume causal relationship have been established. 
• 
Possibly Related: Causal relationship is likely and cannot be excluded.  
• 
Probably Not Related: Good reasons and sufficient documentation not established to assume causal relationship.  
• 
Not Related: The event is most likely related to an etiology other than the trial treatment. 
 
7.3 
Procedure for Reporting an Adverse Event  
Adverse events will be documented during this study by completing the AE Form.  The 
Investigator or designee will sign and date an AE Form for each AE experienced by any subject.  
During the visit, AEs will be evaluated by a member of the study staff.  The nature of each event 
and date of onset, outcome, course, maximum intensity and action taken with respect to treatment 
should be established.  Details of any corrective treatment should be recorded on the AE Form.  
Investigators should follow-up on the status of subjects experiencing an ongoing AE until the event 
has been resolved, or until the condition has stabilized.   
7.3.1. The Investigator or designee will notify the Study Manager or Monitor within 24 hours of 
any Serious Adverse Event that occurred during the study.  The sponsor will promptly review all 
information relevant to the safety of the investigational device. 
7.3.2. The investigator or designee must notify the sponsor and reviewing IRB by phone or fax 
of Unanticipated Serious Device-Related Adverse Events within 24 hours of learning of the event, 
followed by a written a report within 10 working days after learning of the event.   

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7.3.3.   Upon receipt of a report of an UADE by the Ascensia Study Manager or Monitor, the report 
will be immediately forwarded to: 
Andreas Stuhr, MD 
Ascensia Diabetes Care  
Head of Global Medical Affairs 
5 Wood Hollow Road 
Parsippany, NJ  07054 
Phone: 973-906-3613 
e-mail: Andreas.Stuhr@Ascensia.com 
 
7.3.4. The sponsor (Ascensia) must report UADEs to the FDA, all participating investigators, and 
reviewing IRBs within 10 working days after the sponsor first receives notice of the event. 
7.3.5. Regardless of the above definitions, any additional adverse experience that the investigator 
considers serious, and/or of concern in relationship to the study must be documented and reported 
to the Study Manager or Monitor within 24 hours. 
7.4 
Medical Device Reporting 
The Microlet NEXT lancing device system is an Ascensia-marketed product in the US.  If an AE 
occurs that is directly caused by the marketed product, the AE should be reported immediately, 
but in any case within 24 hours, to the Customer Service Department at 1-800-348-8100.  Also 
contact the Ascensia representative listed in Section 12.0.   
8.0 Study Termination and Subject Withdrawal Criteria  
8.1 
Criteria to Withdraw / Discontinue a Subject from Study  
Subjects who experience an AE will be withdrawn from the study.  Subjects may be withdrawn 
from the study at their own request for any reason, or at the discretion of the PI (or designee) for 
one of the following reasons: 
1) 
Illness of a subject 
3) 
Subject non-compliance with protocol requirements 
4) 
Other, at the discretion of the investigator 
In such cases, the subject will be withdrawn from further study participation.  Any evaluable data 
collected from withdrawn subjects will be analyzed and results will be retained for safety 
assessments.  If available, data will be retained in analyses related to the objectives of the study 
unless there is a valid reason to believe that the data may be biased, incorrect, or confounded.  

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9.0 Regulatory  
9.1 
Regulatory Status of the Investigational Devices 
In addition to the regulations outlined in the Ascensia Diabetes Care Global Clinical Affairs 
standard operating procedures, this protocol also complies with the clinical trial regulations of 
British Standard BS EN 13612:2002, “Performance evaluation of in vitro diagnostic medical 
devices.” 
The L-Plus meters, which is the focus of this study, is an investigational devices. For studies 
involving an investigational device, the sponsor is responsible for determining whether submission 
of an IDE application to FDA is required before a study may proceed. The IDE regulations (21 
CFR 812) describe three types of device studies: significant risk (SR), nonsignificant risk (NSR), 
and exempt studies.  
Ascensia has determined that the L-Plus study is a Non-Significant Risk (NSR) study 
which  must follow the abbreviated IDE requirements of 21 CFR 812.2(b), including informed 
consent and IRB review, and do not require submission of an IDE application to FDA. As a result 
of this conclusion, Ascensia has not had correspondence with the FDA regarding determination of 
study classification and risk level. 
Determination of the NSR classification is based on the following: 
• L-Plus  meters are not intended to be implantable devices.   
• L-Plus  meters are not purported or represented to be for use in supporting or sustaining 
human life  
• L-Plus  meters are not intended to provide substantial importance in diagnosing, curing, 
mitigating, or treating diabetes, or otherwise preventing impairment of human health. 
• L-Plus meters do not present a potential for serious risk to the health, safety, or welfare of 
a subject. 
 
9.2 
Institutional Review Board (IRB) Approval  
Prior to study initiation, an IRB must review this protocol, the ICF, and any other supporting study 
documents which impact subject safety.  The investigational site may not begin the study until the 
IRB has given its written and dated approval via a letter that identifies the version/date of the 
protocol and ICF.  A copy of the IRB approval letter and approved ICF must be provided to the 
Investigator and to Ascensia prior to the Study Initiation Visit. 
9.3 
Informed Consent Requirement  
Each subject must provide informed consent before she/he can participate in the study.  The 
informed consent process fully apprises the subjects of the risks and benefits to them and to society 
for participating in the study.  The ICF will clearly state that designated study personnel will be 
able to view the subject’s medical records.  The ICF will also state that Ascensia representatives 
may observe some subjects as part of study staff training, study monitoring or for troubleshooting 
problems with the investigational devices.  

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If the subject understands and agrees to participate in the study, she/he will sign the ICF.  All 
subjects will be given a signed copy of the ICF.  If a subject has a question about his/her rights, 
he/she may contact a member of the IRB at any time during or after study participation. 
9.4 
Study Documentation Procedures 
The investigator will keep study records for a minimum of three years.  Alternatively, other 
arrangements may be made with Ascensia for study document storage.  
9.5 
Study Monitoring   
The study will be monitored by Ascensia representatives.  A monitoring plan will be completed 
by the Study Manager/designee prior to the study.  The Study Manager or designee will conduct a 
study initiation visit combined with 1st monitoring visit, up to 2 interim monitoring visits and a 
close-out visit.  Note: the latter two visits may occur within a continuous timeframe.  
Study monitoring will be performed on site as well as by remote monitoring as per the monitoring 
plan. Per site/sponsor agreement, selected source documents will be scanned and sent to the 
sponsor for remote monitoring by the sponsor. Some of the critical data points that will be 
monitored 100% include, but are not limited to:  
• Meter results 
• YSI control and subject plasma results 
• Centrifugation times 
Ascensia representatives may observe some subject visits as part of study monitoring.  This will 
be done under supervision of the investigator.  Ascensia representatives will maintain subject 
confidentiality and will not interfere with rights of human subjects, safety or bias study conduct.  
9.6 
Investigator’s Report of Study Closure 
Ascensia representatives will send a letter to the site informing them that the study is closed.  The 
study will be considered closed when the data has been locked for data analysis.  
The Investigator or designee will submit a report summarizing subject disposition and other study 
details, as appropriate, to the Ascensia Study Manager and the reviewing IRB.  This report will be 
completed within 3 months of the study closure date.   
In addition, the Ascensia Study Manager, or designee, will report the completion of the study to 
the IRB within 6 months of study closure.  
 
9.7 
Registration of Trial 
The clinical trial will be registered with www.clinicaltrials.gov as required.  
 
9.8 
Clinical Study Report (CSR) 
One CSR will be written by the sponsor for L-PLUS: this report will evaluate data according to 
ISO 15197:2013 criteria.    

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10.0 Data Collection and Management 
A unique number will identify each subject.  The unique number will be the only identifying 
information entered.  A master list of subject names, with their subject IDs, will be kept by the 
Investigator at the study site.  The Investigator will retain all signed IC documents. 
Clinical data will be primarily collected through an EDC system used by Ascensia.  Study data 
will be recorded on forms that will serve as source documents for the EDC system.  These original 
Forms will be retained by the site. 
In addition to data collection, the EDC system will be used for data cleaning as well as monitoring 
operations.  Site and sponsor users will be trained on the system prior to the study start, and their 
access to EDC system will be contingent upon successful completion of training requirements. 
Study personnel will complete and sign all appropriate source document forms in compliance with 
Good Clinical Practice (GCP).  Source documents (Forms) should be completed legibly, in black 
or blue ink.  If it is necessary to make corrections, a single line should be drawn through the original 
entry, the new entry written in, and the correction initialed and dated by the individual making the 
correction. 
The original forms will be retained by the Investigator for a minimum of three years.  
 
11.0 Data Analysis 
The statistical analysis plan for this study is described in document GCA-SAP-2019-002-01. 
In general, meter results satisfying the pertinent objective criteria will be considered “successes” 
in the Bernoulli sense. 
All fingerstick and AST meter test results will be compared to the capillary YSI laboratory results.  
All venous blood meter test results will be compared to the venous blood YSI laboratory results.  
 
 
11.1  Sample Size Considerations 
 
Let MeterBG = meter blood glucose result; 
LabBG = laboratory method (YSI) comparator blood glucose result. 
 
D = MeterBG – LabBG 
RD = 100*(MeterBG – LabBG) / LabBG 
 
Assume 100 evaluable results from people with diabetes.  The ISO 15197:2103 acceptance 
criterion is that 95% of those evaluable results must satisfy the accuracy criteria: 

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|D| = |MeterBG – LabBG | ≤ 15 mg/dL, for LabBG < 100 mg/dL 
or 
 
|RD| = 100*|MeterBG – LabBG| / LabBG ≤ 15%, for LabBG ≥ 100 mg/dL. 
 
With n = 100, Xc = 95 results would be required to satisfy the accuracy criteria.  At P0 = 97.35%, 
there is approximately a 95% chance of satisfying the ISO objective.  Conversely, at Pa = 89.78%, 
there is approximately a 95% chance of failing to satisfy the ISO objective. 
 
11.2 Data Listing  
A listing of the data (excel sheet) is needed for the CSR.  The listing should include:  subject and 
staff fingerstick meter results, AST (palm) meter results, venous meter results, capillary and 
venous mean YSI results, subject ID, hematocrit. 
A listing of subject comments including subject IDs from the Ease of Use Questionnaire is needed 
for the CSR.   
Note: If supplemental or unplanned analyses are requested, they will be presented in a 
supplemental report.   
 
12.0 Administration 
All investigator and site staff communications regarding the study should be directed to the 
Ascensia Study Manager.  Please contact the Ascensia Study Managers noted below for any 
questions or problems concerning the clinical trial. 
 
Maria Teresa Viggiani, MS, CCRA 
Phone: 914-296-2876   Cell: 914-673-7675 
Fax: 1-914-462-4285@efaxsend.com 
e-mail: Maria.Viggiani@Ascensia.com 
 
AND 
 
Pragathi Shelat, B.Pharm, MS 
Phone: 862-229-6679  Cell: 516-313-5289 
e-mail: Pragathi.Shelat@Ascensia.com 
 
 

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13.0 Sponsor Regulatory Binder 
Documents to be collected prior to the start of the study include, but are not limited to: 
1) CVs of Investigator and Sub-Investigators  
2) Signature of PI on page 2 of protocol 
3) IRB approval letter 
4) IRB-approved ICF 
5) Financial Disclosures for PI and Sub- Investigators  
6) Clinical Trials Agreement (including Investigator Agreements for PIs and Sub-Investigators) 
7) Site staff GCP training documentation  
Data / documents to be collected during or after completion of the study include, but are not 
limited to: 
1) EDC database  
2) Progress Notes 
3) Signature and Delegation Log (for all study staff) 
4) Investigator’s Report of Study Closure and IRB acknowledgement 
5) Study-related training documentation 

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Appendix A - Instructions for Cleaning and Disinfecting Meters   
Single-use items, such as test strips, test strip vials or lancing devices, shall not be used with 
multiple subjects and will be discarded in biohazardous containers after use.  Blood glucose meters 
will be for single subject-use, and will be cleaned and disinfected before returning to Ascensia by 
study staff only.  Study staff will disinfect meters using the EPA-registered disinfectant noted 
below that is suitable for use on hard, non-porous surfaces.  Ascensia will provide the disinfectant. 
  
EPA Registration 
Manufacturer 
Product Name 
Active Ingredient 
67619-12 
Clorox Professional 
Products Company 
Clorox (Bleach) 
Germicidal Wipes 
0.55% sodium 
hypochlorite 
 
1. Follow the manufacturer’s directions printed on the container for handling the Clorox 
Germicidal Wipes.  Wear gloves.  
2. Before disinfecting, carefully clean the meter with germicidal wipes until all soil is removed 
and dry as necessary with a clean paper towel.  
3. Note: For proper disinfection, you must keep all meter surfaces wet for 60 seconds.  
4. Using a new germicidal wipe, carefully wipe all outer surfaces of your meter until wet.  
5. After wiping for 60 seconds, use a clean paper towel to dry the meter surfaces and the test strip 
port.  
6. Dispose of all materials used in the cleaning and disinfecting procedure according to current 
laboratory biohazard procedures. 
 
References 
 Protection of laboratory workers from occupationally acquired infections; approved guideline – 
third edition.  CLSI document M29-A3.  ISBN1-56238-567-4. 
 Rutala W, Weber D, Healthcare Infection Control Practices Advisory Committee.  Guideline for 
disinfection and sterilization in healthcare facilities, 2008.  CDC. 
FDA, Processing/Reprocessing Medical Devices in Health Care Settings: Validation Methods 
and Labeling, May 2, 2011. 

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Appendix B -  Instructions for Collection/Processing of Blood for YSI  
Capillary Blood Collection 
1. The study staff will perform a deep finger puncture using a Tenderlett, or similar, lancing 
device, on the subject’s finger.  
2. The study staff will express the blood and collect approximately 200 - 300 µL in a blood 
collection tube (e.g. BD Microtainer with lithium heparin).  It is optional that a hand warmer 
may be used to improve blood flow and facilitate sample collection.  Detailed instructions are 
as follows: 
• Typically the best finger to choose is the middle finger.  It is recommended to practice the 
finger massaging technique prior to puncture. 
• If a hand-warmer is used:, give subject a hand-warmer to hold for several minutes prior to 
puncture.  Note that the hand-warmer should be moved to the subject’s wrist when ready 
for puncture and kept there during the whole collection procedure. 
• Angle finger with tip slightly downward and squeeze fingertip gently prior to puncture. 
• Consider placement of finger puncture device:  Choose a puncture site that will allow you 
to use your dominant hand to collect the blood.  The device should be aligned so that the 
opening (where the blade comes out) is perpendicular to the grooves of the fingerprint and 
so that the opening is placed to the side of the finger (not too close to the nail). 
• While continuing to hold the fingertip, squeeze firmly, hold device firmly against side of 
finger and trigger the blade.  Then turn the finger so that the nail is facing up. 
• Remove pressure from fingertip (to allow capillaries to fill with blood) and collect the first 
drop. 
• Begin massaging motion – continuous message of sides of the finger – this is critical. 
• Continue massaging finger from base to tip as you continue to collect the blood drops into 
the tube.  If blood flow reduces it may help to press on the nail a bit in between massaging 
the finger.  Having the subject stand up and drop their hand may also help. 
 
Processing of Capillary or Venous Blood for YSI Analysis  
1. Once collected, process the microtainer tube for laboratory glucose analysis as follows:   
• Label all tubes with the subject number and a C or a V (Capillary or Venous) as appropriate 
using a Sharpie marker.   
• Gently invert the microtainer tube several times to mix the anticoagulant.  
• Centrifuge the whole blood immediately to separate the plasma from the red blood cells.  
Centrifugation must occur within 15 minutes of first meter blood test.  Timing of all meter 
testing/centrifugation will be recorded on the appropriate CRF.   
• It is recommended that after centrifugation, the plasma is transferred from the microtainer 
tube to a micro/dispo tube or similar container.  Ensure the container is labeled and the cap 
is secured.  
• The capped plasma sample may be kept at room temperature for up to 4 hours before 
testing. 

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2. If testing on the YSI Analyzer is not completed within 4 hours of sample collection, the plasma 
should be refrigerated for up to 24 hours, after which it should be frozen until the sample is 
tested. 
3. Shipment of samples to Ascensia for glucose determinations may be recommended if YSI 
Analyzers do not meet the accuracy guidelines for the Ascensia traceability control sera. 
Arrangements will be made to ship the samples with dry ice via prepaid express mail should it 
become necessary. 
4. Glucose values of the YSI replicates (controls and plasma) should be within 4 mg/dL or 4% of 
each other; if not, an additional assay should be run.  
5. Insufficient (or suspected to be insufficient) samples, missing samples, and other sample issues 
should be recorded in the CRF and on the YSI printout as appropriate.  If a sample is 
hemolyzed, record in the CRF.   

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Appendix C - Instructions for Hematocrit Sample Preparation & Testing  
 
Staff performing the hematocrit will be trained in this procedure during the initiation visit by the 
study manager or designee.    
1. Two hematocrit tubes will be obtained from each subject’s blood.  One tube will be measured 
and the result recorded on the form.  The second tube will be measured only if the first tube 
was not adequate for measurement.  Note: 
• Sample that is collected from venous blood will be collected in un-treated micro-
hematocrit tubes.   
• Samples collected from capillary fingerstick blood will be collected in treated micro-
hematocrit tubes.    
2. Place a microhematocrit tube against the surface of the blood drop.  Do not place the opening 
of the tube flush against the skin or it will be occluded. 
3. Fill the microhematocrit tube about halfway.  Do not over-fill.  Then immediately place one of 
the microhematocrit tube openings in the clay sealant.  Press into the clay 2-3 times to make 
sure enough sealant is transferred. 
4. Place the sealed microhematocrit tube into the micro-centrifuge.  Note: it is very important to 
place the microhematocrit tube in centrifuge rotor with clay side pointed outward.  Spin for 
approx. 2 mins.  Use dialed setting on StatSpin centrifuge.  
5. Reading of hematocrit using StatSpin Crit-reader 
• After collection and centrifugation of hematocrit, place tube rotor in StatSpin Crit-reader, 
with clay side to the right.   
• Slide the marker on the crit-reader to the exact line where the clay and red blood cells meet 
(this is the “0” line).  Press set button until “0” shows in the display.   
• Then move marker all the way over to the other end of the tube to the exact line where the 
plasma ends – press the set button until “100” shows in the display.   
• Now you are ready to read the hematocrit result.  Slide the marker to the exact line where 
the red blood cells meet the plasma. 
• The result in the display is the hematocrit percentage for that sample. 
6. An alternative to Step 5 is manual reading of the hematocrit by the operator using Manual 
StatSpin Micro Hematocrit Capillary Tube Reader.    
7. Record hematocrit result on the appropriate form. 
 
 

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Appendix D - Hypoglycemia Protocol at Rainier 
Rainier Clinical Research Center 
HYPOGLYCEMIA ACTION PLAN 
 
CLINICAL CONDITION 
HYPOGLYCEMIA ACTION PLAN 
• 
Blood glucose <70 mg/dL, but ≥50 mg/dL 
• 
With or without symptoms 
• 
Treat if patient exhibits neuroglycopenic 
symptoms, patient requests or 
investigator discretion 
• 
Treat hypoglycemia with fast-acting 
carbohydrates: 16 grams initially, with 
repeat treatment every 15-20 minutes at 
discretion of investigator until blood 
glucose is increasing or blood glucose 
≥100 mg/dL 
• 
Blood glucose <50 mg/dL 
• 
With or without symptoms 
• 
Subject able to self-administer fast-acting 
carbohydrates 
• 
Treat hypoglycemia with fast-acting 
carbohydrates: 16-32 grams (amount at 
investigator discretion) initially , with 
repeat treatment every 15-20 minutes at 
discretion of investigator until blood 
glucose is increasing or blood glucose 
≥100 mg/dL 
• 
Blood glucose <50 mg/dL 
• 
Subject is NOT able to self-administer 
fast-acting carbohydrates 
Severe hypoglycemia (defined as a hypoglycemic 
event requiring assistance of another individual to 
actively administer treatment) 
• 
Treat hypoglycemia with 1 mg 
intramuscular glucagon or intravenous 
glucose 20cc of 50% glucose  
• 
Monitor vital signs and blood glucose 
until glucose returns to normal range and 
patient regains full consciousness 
• 
If on an insulin pump, suspend insulin 
delivery until glucose is ≥70 mg/dL 
• 
Transfer to acute care facility at 
discretion of the investigator 
 
 
 
 

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 Attachment 1:   Proper Handling of Ascensia YSI Serum Controls 
 
 
Ascensia R&D completed characterization of six-level serum controls purchased from Bio-
Techne® (formerly Bionostics).  During the characterization, staff observed that sample 
stratification during thawing can occur if the control vials are not handled judiciously.  The 
recommended procedure is as follows. 
 
1. Thaw the frozen control amber glass vials at 2-8ºC for 2-3 days in the refrigerator.  Each vial 
holds approximately 2.0 mL. 
2. Before opening, mix the vials well by gently inverting them at least 10 times. 
3. Prepare an aliquot of each level in capped microcentrifuge tubes.  Cap tightly.  We prefer 
Fisherbrand™ Microcentrifuge 0.5mL tubes (PN 02-681-333) with O-ring seal caps (PN 02-
681-358). 
4. Store any remaining control in the amber glass vials at 2-8ºC. 
5. Before running a control sample on the YSI, mix the aliquot tubes by gently inverting them. 
6. At the beginning of each day, remix the amber glass vials and refill the aliquot tubes.  When 
not in use, store the aliquots at 2-8ºC. 
7. The thawed control use life is two weeks.  Discard all thawed control—aliquot tubes and amber 
glass vials—after two weeks. 
8. Since slow thawing for 2-3 days is recommended, it is advisable to keep an extra thawed set 
available for testing.