bluuebunny/crossref_metadata_2025 huggingface dataset:
A-101 Is That Unexpected? Education on Handling Real World Complications with Tightly Controlled Analytes During Method Evaluation
Authors: Love S A , Aguanno J , Melchior J , Burnett R , Muldowney T , Von Rotz A , Guerrero A , Pollan M
Published: October 2024

Background

A common activity in clinical laboratories is performance evaluations. Given the importance for a new analyzer being put into clinical use, this exercise typically requires extensive preparation with significant time and resources invested to bring on new instrumentation. During early adoption, studies examined real-world performance and comparability of the Atellica® CI Analyzer (Atellica CI) to clinically used instruments. Precision, linearity, and method comparison (MC) studies, conducted in accordance with CLSI protocols, were completed at several locations installing new instruments to examine the analytical performance of common clinical laboratory assays. This work examines the impact of using only unmodified specimens to verify performance during method comparison for tightly controlled biological analytes like electrolytes and illustrates the impact of the tested sample range on regression analysis.

Methods

Testing was completed using unmodified, de-identified, remnant patient serum specimens. The MC studies for these electrolytes were completed over multiple days using single replicates from at least 40 individual specimens, on two instruments, the Atellica CI and the Dimension® EXL in accordance with CLSI EP09-3A. The effect of sample range on regression was demonstrated through data restriction and inclusion of modified sample results within the data analysis. Data analysis was completed using Analyse-it for Microsoft Excel (version 6.15.4).

Results

Representative method comparison data for sodium, chloride, and potassium found slope differences were more pronounced when sample concentrations were more limited. Individual assay slopes were determined using Deming linear regression model and comparisons between restricted samples show the effects of sample range on regression and correlation parameters.

Conclusions

Understanding the relationship between sample range and regression analysis is helpful to illustrate an important fundamental concept of method comparison. These examples illustrate the importance in covering as much of the analytical measuring interval as possible and the impact on regression analysis when samples are narrowly grouped.
A-104 Study of the matrix effect for thyroglobulin and calcitonin in fine-needle aspiration of cervical nodules
Authors: Canali D , Zampieri G , Bonato C , Francisco G , Sabino C , Oliveira V , Lopes A
Published: October 2024

Background

Analysis of material from fine needle aspiration of cervical thyroglobulin nodules with Beckman Coulter reagents on the ACCESS equipment and Calcitonin using the Siemens reagent on the Immulite equipment. The matrix effect, linearity and the limit of analytical sensitivity were analyzed. This study aims to analyze the correlation between the serum/plasma results described in the suppliers' package inserts with the results obtained with the puncture matrix.

Methods

The performance characteristics were tested according to the laboratory's operating procedures, based on the recommendations of ANVISA RDC 166/2017. Such parameters involved doping, which corresponds to the introduction of a calibrator with a known and traceable value into a pool of puncture material that is known to be negative, in order to analyze the behavior of the results obtained, paying attention to the deviations that may eventually occur due to the interference of the characteristics of the puncture material. puncture. After doping the material, dilutions were performed up to the sensitivity limit of the test, also analyzing the functional sensitivity, where ten replicates of the material were dosed, thus obtaining CV%, mean and Standard Deviation values. The same process was carried out and evaluated using the serum material. During the entire process, the materials received and analyzed were centrifuged and always kept in a refrigerated environment.

Results

The recovery% of the matrix effect study for cervical aspirate puncture showed results within the expected range, ranging from 80% to 120%, as well as the results compared with serum/plasma referenced in the supplier's package insert. The functional sensitivity study for Thyroglobulin showed a CV% of 1.77%, being lower than that recommended by the package insert (2.2%) and for the Calcitonin analyte, the functional sensitivity study showed a CV% of 8.49%, being lower than recommended by the package leaflet (15.7%), in addition to having CV's lower than the 20% recommended by ANVISA. The results obtained with the matrix effect study for serum also showed values between 80% and 120%, both of which agreed.

Conclusions

The results showed excellent performance for material from fine needle aspiration for both Calcitonin and Thyroglobulin, being consistent with the serum data described by the suppliers' package inserts, thus making the reagent usable for the cervical aspirate puncture matrix.
A-105 Cherry-red plasma: unraveling the impact of PV-10 on chemistry assays
Authors: Chen L , Zhang Y , Meng Q
Published: October 2024

Background

The accuracy of clinical laboratory measurements can be compromised by laboratory test interferences. PV-10 (Rose Bengal), a chemotherapy medication used in the treatment of metastatic melanoma, was observed to cause a cherry-red discoloration in plasma. This plasma color change, resembling hemolysis, raises serious concerns about potential interference in clinical laboratory tests. Our study focused on understanding the potential interference of PV-10, particularly the impact on the hemolysis index (H index) and various chemistry assays to ensure result accuracy in cancer treatment.

Methods

To investigate the potential interference of PV-10, pooled normal and high baseline bilirubin plasma samples were spiked with a gradient of PV-10 concentrations (0 to 22 µg/100µL). Samples were analyzed on Roche Cobas 8000 analyzer with H index and all the chemistry assays measured.

Results

The presence of PV-10 in plasma significantly increased the H index, in a dose-dependent manner. Specifically, the H index increased from 14 (control plasma) to 2643 at the highest PV-10 concentration (22 µg/100µL) in pooled plasma. Among all chemistry assays, most of them were not affected by PV-10. However, certain assays like IRON have dose dependent increase with PV-10 concentration. CHO2A (Total Cholesterol), TP (Total Protein), and TRIG (Triglycerides) have shown a consistent increase trend. In contrast, the DBILI values decreased with increasing PV-10 concentrations, from 6.7 mg/dL to 0.2 mg/dL. TBIL levels also demonstrated a gradual decline from 9.6 mg/dL to 8.4 mg/dL as PV-10 concentration increased in pooled high baseline bilirubin plasma.

Conclusions

This study provides insights into the potential interference of PV-10 in clinical chemistry assays. The elevated H index by PV-10 is not attributed to hemolysis. It is imperative for both laboratory professionals and care providers to recognize the implications of PV-10-induced plasma discoloration and its potential interference in laboratory tests.
A-113 Evaluation of Cystatin C Ordering Trends in a Quaternary Care Health System
Authors: Agana B , Ness M , Marzinke M A
Published: October 2024

Background

Cystatin C is an alternative and adjunct kidney biomarker to creatinine in the estimation of glomerular filtration rate (eGFR). The National Kidney Foundation and American Society of Nephrology have called for increased use of cystatin C in the evaluation of kidney function. Our institution brought cystatin C testing in-house in August 2021. The goal of this study was to evaluate the utilization and longitudinal ordering trends of cystatin C within our quaternary care health system.

Methods

Cystatin C is measured on a Roche cobas® c502 using an assay traceable to IFCC-certified reference material. The electronic health record system (Epic) was queried using Epic SlicerDicer to identify ordering trends within our academic medical center between July 2021 and January 2024. The frequency of unique (one-time) cystatin C orders and concomitant orders with creatinine was also conducted. An in-depth analysis and chart review was performed over a one-month period to evaluate ordering locations, underlying clinical conditions, including previous diagnosis of kidney disease, and correlations between cystatin C and creatinine measurements.

Results

Between July 2021 and January 2024, 11,577 tests were performed from 8,415 unique patients. Cystatin C orders remained stable since April 2022, with an average 370 orders (363-378) per month. Eighty percent of cystatin C test requests were accompanied with creatinine testing during the same encounter. In a detailed analysis of a one month period, cystatin C testing was primarily performed in patients with an existing diagnosis of kidney disease (66%) and in the evaluation of renal function in patients with a cancer diagnosis (23%).

Conclusions

There has been sustained adoption of cystatin C by providers at our institution. The majority of orders have occurred in patients with an underlying renal diagnosis, as well as comorbid or chronic conditions that may affect kidney function.
A-116 Pleural Fluid pH - Are We Collecting and Measuring It Correctly?
Authors: Tan C , Tan S , Chiong S , Yeo C
Published: October 2024

Background

Pleural fluid pH has diagnostic, therapeutic, and prognostic implications in the management of pleural space infections and malignancy. Pleural fluid sample collection, transportation and processing are known for source of pre-analytical error which contribute to inaccuracy of measured pH with clinical implication to patient care. Some clinical practices observed locally include pleural fluid collection in sterile container for ease of other biochemistry analytes measurement in addition to pH. This study aimed to compare the differences in pleural fluid pH and other commonly ordered biochemistry analytes collected using sterile container against heparinized blood gas syringe.

Methods

Total 20 Pleural fluid samples were collected from patient using a procedure called thoracentesis from, with approximately 2 to 3ml of fluid directly transferred into heparinized blood gas syringe as well as in a sterile container. Both samples were then transported to Clinical Biochemistry Laboratory within an hour. Both samples were analyzed on 3 analytical systems; 1) pH on Roche COBAS b221 blood gas analyzer, 2) Glucose, Total Protein, LDH on Beckman AU System and 3) Specific Gravity on Reflectometer. Statistical analysis was performed using Analyze-It software.

Results

pH fluid collected in sterile container demonstrates significant high bias (absolute of 0.4 units or 5%) as compared to sample collected in heparinizes blood gas syringe. Other commonly ordered tests in fluid profile (Glucose, Total Protein, LDH and SG) demonstrate good correlation (Altman-Bland Difference Plot <5%) for pleural fluid collected in sterile container as compared to heparinized syringe.

Conclusions

This positive bias observed is likely attributed to air exposure in pleural fluid collection in sterile container as compared to heparinized syringe. The accuracy of measured pleural pH is critically dependent on the method of sample collection. This study highlights the need for a standardized protocol for pleural pH collection for accurate measurement.
A-120 Analytical and Clinical Verification of the Elecsys® Alzheimer’s Disease Cerebrospinal Fluid Biomarkers pTau/Abeta42 and Total-Tau/Abeta42
Authors: Troike K , Zilka S , Bekris L M , Leverenz J , Khrestian M , DiFilippo F P , Lowe M , Colón-Franco J M
Published: October 2024

Background

Alzheimer’s disease (AD) is the most common form of dementia characterized by amyloid beta (Abeta) plaques and neurofibrillary tau tangles in the brain. Cerebrospinal fluid (CSF) biomarkers Abeta42, total tau (tTau), and phosphorylated tau (pTau181) offer a high degree of sensitivity and specificity in the diagnosis of AD and have utility in monitoring disease progression. AD biomarker ratios, pTau181/Abeta42 and tTau/Abeta42, correlate with amyloid positron emission tomography (PET) imaging, and several assays have received FDA clearance. This study was designed to evaluate the analytical performance and clinical utility of AD-related Abeta and tau biomarkers on the Roche cobas e601.

Methods

Precision studies were performed using Roche PreciControl kits. Intra- and inter-precision samples were tested ten times in a single day or twice a day for ten days, respectively. Five levels of Roche Calcheck material were measured in triplicate for linearity. Method comparison was carried out using 39 samples from the Elecsys CSF Correlation Sample Panel from Roche. Qualitative comparisons to amyloid PET Centiloid scores were performed using 35 CSF samples (n=11 PET negative, n=24 PET positive). CSF samples and PET interpretation were obtained from the Cleveland Clinic Institutional Review Board approved Cleveland Alzheimer's Disease Research Center (CADRC) and Cleveland Clinic Lou Ruvo Center for Brain Health Aging and Neurodegenerative Disease Biobank (LRCBH-Biobank).

Results

Conclusions

Evaluation of AD biomarkers on the Roche cobas e601 revealed reliable precision and analytical accuracy. The pTau181/Abeta42 and tTau/Abeta42 ratios demonstrated excellent positive agreement when compared to amyloid PET, but poor negative agreement. This may be due to the small sample size of PET-negative patients. Alternatively, as amyloid PET detects neuritic plaque accumulation, the CSF biomarker-positive/PET-negative patients may have early non-neuritic amyloid plaque pathology not detectable by imaging. Overall, this assay is suitable for use and may serve as a valuable tool in AD diagnosis.
A-126 The Role of Lysozyme and Other Inflammatory Markers in Understanding GI Microbiome Changes
Authors: Reynolds D , Gruzdys V , Mortensen J E
Published: October 2024

Background

The role of GI microbiome in gut health, intestinal permeability, and the development of inflammation has been well-established however, few clinical studies assess microbiome (i.e. PCR) and biochemical markers from the same stool collection. Doctor’s Data Inc. (DDI) has been performing a GI microbiome test in conjunction with other stool biochemical markers for the past 3 years, with over 7000 patient samples analyzed. The aim of this study was to analyze GA-map® Probe results for possible correlations with elevated values for fecal calprotectin, lactoferrin, and lysozyme.

Methods

Microbiome characterization The microbiome analysis test used in this study identifies 48 targeted analytes across six phyla using the GA-map® Dysbiosis test (Genetic Analysis AS, Oslo). The test algorithm has been trained and validated using 48 bacterial markers signal intensity obtained from 668 adults, including healthy subjects, patients with IBS and IBD. GA-map® Dysbiosis Analyzer software calculates the degree of dysbiosis (DI) of a given sample, assigning a DI ranging from 1 to 5. DI greater than 2 indicates that the microbiota is deviant from a healthy reference population (“dysbiotic”), while a score of 2 or lower is reported as “normobiotic”. Biochemical marker characterization Each stool sample was analyzed for a suite of biochemical markers - in particular calprotectin, lactoferrin and lysozyme - with the concentration determined using commercially available ELISA kits (ALPCO, Techlab).

Results

Using clinical cut-offs of 100.0 ug/g (calprotectin) and 7.3 ug/g (lactoferrin) the patient data was sorted into cohorts indicating disease state of “Normal”, “Intermediate” and “IBD Likely”, and further differentiated using the lysozyme result. The biochemical marker results were correlated against DI score for each stool sample (Figure 1).

Conclusions

DI score increases as biochemical inflammatory markers increase, with lysozyme as the key differentiating factor between patient cohorts.
A-129 Evaluation of the Analytical Performance of Magnesium and Calcium General Chemistry Assays on the Atellica CI Analyzer
Authors: Carter R H , Varhue W , Gupta P , Shalhoub V , Camacho J , Snyder J
Published: October 2024

Background

The Atellica® CI Analyzer is an automated, mid-throughput, integrated chemistry and immunoassay analyzer utilizing both Atellica® CH and Atellica® IM assays. This study was designed to evaluate the analytical performance of the Atellica® CH Magnesium (Mg) and Calcium (Ca), Assays on the Atellica CI Analyzer.

Methods

The Atellica CI Mg and Ca assays use the same reagents and calibrators as the Atellica CH assays. Precision studies were performed according to CLSI EP05-A3 using serum and urine samples. One aliquot of each sample pool was tested in duplicate in two runs per day >2 hours apart on each analyzer for 20 days. MC studies were performed according to CLSI EP09-A3. Individual human urine and serum samples were analyzed using the Atellica CH and Atellica CI Analyzers. LoB, LoD, LoQ were determined according to CLSI EP17-A2.

Results

Representative precision and MC results observed across indicated sample ranges are listed for each assay in the table below. Over the two assays tested, repeatability and within-lab %CVs were <4.9%. Overall, slopes determined using the Deming linear regression model were approximately equal to 1. LoB, LoD, LoQ for serum Mg were 0.07 mg/dL (0.03 mmol/L), 0.11 mg/dL (0.05 mmol/L), and 0.50 mg/dL (0.21 mmol/L), and for urine Mg, 0.07 mg/dL (0.03 mmol/L), 0.14 mg/dL (0.06 mmol/L), and 1.00 mg/dL (0.41 mmol/L). LoB and LoD for serum Ca were 0.2 mg/dL (0.05 mmol/L) and 0.3 mg/dL (0.08 mmol/L), and for urine Ca, 0.3 mg/dL (0.08 mmol/L) and 0.5 mg/dL (0.13 mmol/L), respectively. Linearity was demonstrated for both assays.

Conclusions

Atellica CH Mg and Ca Assays on the Atellica CI Analyzer demonstrated good precision and equivalent performance to the same assays on the Atellica CH Analyzer.
A-130 Intercomparison of Methods between Atellica Solution® and CI® Analyzers for Alanine Aminotransferase (ALT) and Aspartate Aminotransferase (AST)
Authors: Barrionuevo-Gonzalez M , Lapeña-Garcia S , Garcia-Valdecasas S , Costales-Lucia T , De La Rubia-Maestu J , Ruiz-Alvarez M
Published: October 2024

Background

Objectives: To verify that the results of ALT and AST measured by the Atellica Solution® analyzer (and the new CI® analyzer (Siemens Healthineers) are interchangeable.

Methods

For each assay, 40 patient samples were selected, covering the entire measurement range, and analyzed by both instruments. To determine the degree of agreement between the instruments, Bland Altman analysis of mean differences and Passing Bablok regression were performed. Statistics are expressed with their 95% confidence intervals (CI). The evaluation of results was carried out using the statistical program Method Validator.

Results

ALT: In Bland-Altman analysis, no constant systematic error was found as the 95% confidence interval of the mean difference includes zero. Similarly, in Passing Bablok regression analysis, the confidence interval of the intercept includes zero. No systematic proportional differences were observed based on the slope, as the 95% confidence interval includes the value one. AST: In both Bland-Altman and Passing Bablok regression analyses, a constant systematic error was detected as the 95% confidence interval of the observed mean difference Bland-Altman and the 95% confidence interval of the intercept of the regression line do not include zero. A slight systematic proportional difference was also observed based on the slope, as the 95% confidence interval does not include one.

Conclusions

Based on the observed data, we can conclude that the Atellica Solution® and CI® analyzers are interchangeable for the ALT assay. There are constant systematic differences and a slight proportional systematic error for the AST assay, which should be re-evaluated after a new calibration of the method. It is noteworthy that this variation in the data is more pronounced for high AST values, not being clinically significant differences at these concentrations; nor as pronounced in clinical decision values. Therefore, we can conclude that the Atellica Solution® and CI® analyzers are interchangeable for AST.
A-143 Comparative Evaluation of K2EDTA Performance in Multinational Vacutainers: A Case Study of “DBO” by Schraubg and “BD” by Becton, Dickinson
Authors: Patel T , Patel K , Raj D , Bapu R
Published: October 2024

Background

This research paper presents a thorough comparison of the anticoagulant properties of K2EDTA in two multinational vacutainer brands, “DBO” by Schraubg and “BD” by Becton, Dickinson, with a specific focus on critical hematological parameters. The study encompassed the analysis of 50 samples, spanning a wide range of values from 60 fl to 110 fl, evaluating key parameters such as Hemoglobin (Hb), Hematocrit (HCT), Mean Corpuscular Volume (MCV), Mean Corpuscular Hemoglobin Concentration (MCHC), and Red Blood Cell (RBC) count.

Methods

Peripheral blood samples were gathered from 50 outpatient participants using the specified tubes. The samples underwent assessment for six hematological analytes, including RBC Count, hemoglobin, HCT, MCV, MCH, and MCHC. Comparative analysis was performed using the paired t-test. Differences exceeding the total allowable error were considered clinically significant. To assess stability, initial results were compared with those obtained from samples preserved for a duration of 2 hours

Results

Our findings indicate that the use of “DBO” vacutainers resulted in slightly higher values for Hb, HCT, MCV, MCHC, and RBC count compared to the “BD” vacutainers. Notably, the statistical significance was observed in MCV, HCT, and MCHC across nearly all samples, while Hb and RBC parameters showed no significant changes. Given the crucial role of K2EDTA as an anticoagulant in maintaining the hypotonicity of Red Blood Cells (RBCs), ensuring accurate hematological analyses is imperative.

Conclusions

Although the T-value for Hemoglobin did not exhibit statistical significance, the study identified notable differences in Hematocrit (HCT) and Mean Corpuscular Volume (MCV). This underscores the importance of selecting the appropriate vacutainer brand to ensure reliability and accuracy in hematological measurements, particularly when assessing MCV, where even slight variations can carry clinical implications. The results underscore the necessity for clinicians and laboratories to carefully consider the choice of vacutainers based on their specific requirements and the critical nature of the hematological parameters under analysis.
A-150 Extended Pre-Analytical Stability Validation for Coagulation Tests in Brazilian Clinical Laboratory
Authors: Canali D , Fijita A , Sluzula M , Vieira L , Nakatani S , Colombini M , Silva C , Dias C , Lopes A
Published: October 2024

Background

The transport of samples for coagulation tests is a challenge for clinical laboratories, due to their limited stability and short time for processing according to the manufacturer's recommendations. The objective of this study was to evaluate the possibility of extending the stability of samples collected for Prothrombin Time and Activity (PTA), activated Partial Thromboplastin Time (aPTT) and Fibrinogen tests for up to 10 hours at room temperature, in comparison with the stability described in the literature and by the manufacturer of the reagents (4 hours for aPTT, 8 hours for PTA and Fibrinogen). We verified the acceptability of the samples with a longer period between collection and analysis, including its impact on the clinical management

Methods

Four tubes of 3.2% sodium citrate from 40 volunteers were analyzed at times of up to 4 h, 6 h, 8 h and 10 h. The results obtained were compared with the dosage performed within 4 hours (recommended by manufacturer) and the Total Error obtained in the analysis of the samples was evaluated. The tubes remained at controlled room temperature of 15 to 25°C and were centrifuged at 1500 g for 15 minutes. Then, the analyses were performed by coagulometric method.

Results

The results showed safety in the analysis of samples for coagulation tests even within 10 hours after collection at ambient temperature and were within the Maximum Allowable Total Error (Carmen Ricós): 95% of the PTA results (TE 5,3%); 92.2% of aPTT results (TE 4,5%) and 98% of Fibrinogen analyses (TE 13,6%) (Figures 1, 2 and 3). In addition, even the results that exceeded the specified Total Error showed no impact on the clinical decision.

Conclusions

The study demonstrated that there was no significant loss of stability for the analyses of PTA, aPTT and Fibrinogen performed on samples kept at room temperature within 10 hours after collection. Most of the results showed variation within the specified Total Error and all results showed acceptability from a medical point of view
A-158 Multi-site verification and performance of body fluids in-between Horiba Medical Yumizen h2500 with Sysmex XN-10 automated hematology analyzers and manual microscopy for morphology assessment
Authors: Hege M L
Published: October 2024

Background

The analysis of body fluids plays an important role in the diagnosis and management of a wide variety of conditions. As standards for method validation and laboratory developed tests evolve, the body fluids analysis is receiving increased attention from both laboratories and regulatory bodies. We present the evaluated result of the validation and clinical performance accuracy of HORIBA Medical Yumizen H2500 analyzers as compared to SYSMEX XN-10 analyzers and to manual slide microscopy examination as a reference method for differential analysis for body fluids samples.

Methods

Depending on each laboratory collected procedures, synovial samples were collected with Sodium Heparin or K2EDTA or without anticoagulant and the other samples were collected with K2EDTA or without anticoagulant. As recommended, most (94%) of the synovial fluids were treated with the hyaluronidase. For the comparison of Yumizen H2500 versus XN-10 analyzers, the study was performed in 2 US sites and 1 French site collecting 390 samples totally: 138 serous fluids (Ascites, Pleural, Peritoneal, Pericardial), 169 synovial fluids and 83 CSF fluids. Samples were analyzed as soon as possible after collection (with a limit of 24 hours for serous and synovial fluids and 4 hours for CSF) and within 2 hours between both instruments. For the comparison of Yumizen H2500 versus reference method (manual microscopy), the study was performed in 2 US sites collecting 58 samples: 34 serous fluids (Ascites, Pleural, Peritoneal, Pericardial), 12 synovial fluids and 12 CSF fluids. Two qualified morphology examiners made the manual microscopy analysis and the average of the results of the 2 slide readings was used for the analysis.

Results

Considering all types of body fluids, the H2500/XN results are very well correlated for BFWBC (0.969), BFRBC (0.999), BFMN% (0.912), BFMN# (0.89), BFPN% (0.912), BFPN# (0.97). Similarly, the statistical analysis per body fluids type shows very good correlation. For serous type, all results are higher than 0.876, for synovial type all results are higher than 0.879 and for cerebrospinal fluids type, all results are higher than 0.788. Among all samples, less than 8 samples (<2%) were out of the 95% interval. The comparison Yumizen H2500 versus manual microscopy, all the correlation results (r²) are very good: 0,9705 for Polynuclear and 0,9742 for Mononuclear. The study demonstrated better results of Yumizen H2500 versus manual microscopy than versus XN-10 for BFPN% (0.9705 versus 0.912) and BFMN% (0.9742 versus 0.912). Among 58 samples, only 4 (6.9%) were out of the 95% interval for BFPN% and BFMN%.

Conclusions

The Yumizen H2500 results for body fluid WBC differentiation (Polynuclear and Mononuclear) are very well correlated with the results from manual slide microscopy evaluation. The comparison of the results analyzed on Yumizen H2500 and on XN does not show any significant difference.
A-171 Novel Fluorometric Assay for Pediatric Anti-Factor Xa Testing: Minimizing Bilirubin and Hemolysis Interference in Whole Blood
Authors: Pamula V , Basmajian M , Ullal A , Nichols L , Adhikari S , Moser M , Emani S , Emani S
Published: October 2024

Background

High bilirubin levels are common in pediatric patients and are noted in patients undergoing extracorporeal membrane oxygenation (ECMO). Acute anticoagulation therapy necessitates frequent monitoring of anti-Factor Xa activity (aFXa), and high bilirubin levels cause interference in chromogenic assays. Moreover, large volumes of blood lead to iatrogenic anemia in pediatric and neonatal patients. We developed a fluorescence assay to measure aFXa activity on a near-patient digital microfluidic (DMF) platform using low volume whole blood samples (< 50 μL) and present the results of interference of bilirubin and hemolysis on aFXa activity.

Methods

aFXa assay was performed on a DMF cartridge wherein 50 μL whole blood was loaded into the cartridge which separates plasma droplets through agglutination within the cartridge followed by incubation with a droplet containing exogenous FXa and a fluorogenic substrate. The fluorescence is inversely proportional to the concentration of heparin in the sample. All required reagents for the aFXa assay are dried within the cartridge, allowing for integrated sample preparation and assay automation for point of care use. To test the interference of bilirubin, we spiked into whole blood different concentrations of unconjugated bilirubin, ranging from 0-40 mg/dL. We also tested hemoglobin interference by testing 4 different concentrations of hemolysate (0-1,000 mg/dL) on the aFXa assay. All experiments were performed with 6 replicates of each concentration.

Results

Across each bilirubin concentration, we saw uniform measurements of aFXa activity (see Table). We observed < 5% CV in aFXa activity with 0, 13.33 and 26.67 mg/dL bilirubin levels. All measured unfractionated heparin values for hemolysate interference were <5% CV.

Conclusions

Our interference results suggest that high bilirubin and hemolysate levels do not interfere with our novel aFXa fluorescence assay. Further clinical studies are underway to establish clinical performance.
A-172 Feasibility of PT, aPTT, and Fibrinogen on a Multifunctional Digital Microfluidic Cartridge
Authors: Pamula V , Ceasrine A , Ruddiman C , Caudle B , Ng R
Published: October 2024

Background

Excessive bleeding is prevalent in severe trauma patients, and measurement of multiple coagulation parameters is vital to correcting coagulopathy and improving outcomes. There are no rapid, near-patient technologies that can perform multiple coagulation assays simultaneously on a single consumable. We are developing a point-of-care digital microfluidic (DMF) platform to test three critical coagulation markers: prothrombin time (PT), activated partial thromboplastin time (aPTT), and fibrinogen.

Methods

Five proficiency testing plasma samples were obtained from the College of American Pathologists (CAP) with normal, intermediate, and prolonged response of PT and aPTT, and normal and elevated fibrinogen levels. After loading 10 μL of sample onto the cartridge, autonomous digital microfluidic operations divided the sample into three droplets, which were then combined with reagents for PT, aPTT or fibrinogen. The viscosity of these droplets, measured from the electrical impedance on a toaster-sized instrument, changes at a rate proportional to the amount of coagulation precursors present from which time/activity was calculated.

Results

Classification of samples into normal, intermediate or prolonged categories along with the respective times obtained from the DMF cartridges matched those obtained on the Stago Compact Max instrument (Figure). We also demonstrated separation of plasma from whole blood within the DMF cartridge using agglutinating reagents, enabling an easy-to-use workflow without the need for centrifugation.

Conclusions

Our preliminary results demonstrate that PT, aPTT and fibrinogen can be measured on a single cartridge with performance similar to comparator methods, with a time to result of <15 minutes. To our knowledge, this is the first time these 3 assays have been performed simultaneously on a single consumable and furthermore, this is the first time viscoelastic tests have been performed on a multifunctional instrument that also performs chemistry and molecular tests. Studies are underway to evaluate analytical and clinical performance.
A-173 Evaluation of Artificial Intelligence (AI)-assisted Flow Cytometry Analysis for Plasma Cell Neoplasms
Authors: Yeager T , Lyu Y , Lu Z , Morita M , Wang Z , Fan G
Published: October 2024

Background

Flow cytometry plays a critical role in diagnosing hematologic disorders, yet manual analysis is time-consuming and prone to variability. Significant advancements in artificial intelligence (AI) within healthcare have occurred in recent years. Previously, we developed and evaluated an AI-assisted automated system for diagnosing acute leukemia. This study aims to assess the effectiveness of the AI system in diagnosing plasma cell neoplasms.

Methods

A total of 76 bone marrow samples were analyzed using a panel of antibodies (CD38, CD56, CD138, Kappa, and Lambda) to determine plasma cell percentages and immunophenotype. All samples were used for initial diagnosis of plasma cell neoplasm or therapeutic monitoring. Manual analysis using Kaluza™ software served as the gold standard. The same flow cytometry data was processed by the AI software (DeepFlow™) for comparison. Pearson correlation coefficients were used to compare AI and manual analysis for diagnosis and plasma cell count.

Results

Among the 76 cases, manual analysis identified 57 positive cases for monoclonal plasma cells (36 cases of CD38+/CD138+/Kappa+ and 21 cases of CD38+/CD138+/Lambda+), with 89% showing aberrant CD56 expression. AI identified 49 monoclonal cases, missing 8 cases with low plasma cell levels (<0.07%) or brighter fluorescence. AI accurately matched 97% of immunophenotypes with manual analysis. Pearson correlation coefficients demonstrated excellent correlation (r=1.0) between manual and AI plasma cell percentages. Furthermore, AI has also enhanced laboratory efficiency, reducing analysis time per case from 10 minutes with manual methods to just 3 minutes with AI analysis.

Conclusions

This study shows promise for AI-assisted flow software in diagnosing plasma cell neoplasms. AI has enhanced operational efficiency by reducing the manual processing and analysis time. Further training is needed to improve the identification of variations in fluorescence intensity and low plasma cell levels.
A-174 Comparison of Bleeding Risk Scales in Elderly Patients With Persistent Atrial Fibrillation
Authors: Menendez Mite A R , Camacho F , Vaca R , Valverde A
Published: October 2024

Background

Atrial fibrillation is a significant cause of thromboembolism and sudden death in the world making anticoagulant treatment a cornerstone; however, it is associated with bleeding risk. Currently, there are no studies in Ecuador that support the prognostic validity of the recent bleeding risk scales, as well as the lack of research to discover novel methods that are applicable and more accessible to use within the medical care setting.The Hemorr2hages, ORBIT and ATRIA scores represent a promising approach for those health centers that do not have INR available and consequently cannot measure risk through HASBLED (internationally recommended score). In this way, hemorrhagic events can be prevented in patients with atrial fibrillation with anticoagulant treatment without exception. Our main goal is to compare the predictive bleeding risk scores of 3 scales (Hemorr2hages, ORBIT, ATRIA) versus traditional HASBLED score.

Methods

In this retrospective cohort study, we assessed chronic persistent atrial fibrillation in 100 patients using the electronic medical records in an outpatient clinic in Guayaquil, Ecuador. We calculated 3 bleeding risk scores for each patient according to the variables measured in each one of them and used those to compare the categories against HASBLED’s. A Pearson correlation index was obtained, and ROC performance curves were made. P value <0.05 was deemed statistically significant.

Results

Of the 100 patients included, 65% were males, and mean age was 75±10.73. We reported only 6% bleeding events during the follow up. According to HASBLED score: 31% were at high risk. In comparison: 40% and 37% were also at high risk in Hemorr2hages and ATRIA scores, respectively. HASBLED was fairly correlated with Hemorr2hages and ATRIA scores (Pearson’s R: 0.617, p:<0.001; R:0.528, p:<0.001, respectively). Only 22% were classified at high risk by the ORBIT score. ROC curve analysis showed ORBIT’s AUC was 0.963, while Hemorr2hages, ATRIA and HASBLED were 0.931, 0.925, and 0.732 (p:<0.001), respectively.

Conclusions

In our sample, ORBIT score was better correlated with bleeding episodes than the other scales. This a promising approach for those health centers that do not count with INR reactive materials available for it to be measured at their laboratories. Further research is needed to accurately analyze whether criterion included in ORBIT score might predict bleeding scores better than HASBLED, Hemorr2hages and ORBIT tools; since the items required to calculate it are more readily accessible.
A-176 Do you have a hemoglobin variant?
Authors: Fan S
Published: October 2024

Background

Hemoglobinopathies are the most common genetically inherited red blood cell disorder. While there are more than 1,000 hemoglobin variants currently identified, the most common ones include hemoglobin S, Hemoglobin C and hemoglobin E. Patients with homozygous variants are usually identified at young age due to symptoms, whereas patients with heterozygous variants (carriers) can be undiagnosed due to clinically benign nature. Although at low risk, extreme conditions such as high-intensity physical activity can lead to serious health issues in sickle cell trait (carriers). Moreover, children from carrier parents may have homozygous conditions that can produce devastating consequences. This study is to find out a rough estimate of patients’ awareness of their own carrier state of a hemoglobin variant in the population of our medical center.

Methods

Hemoglobin A1c (HbA1c) is performed on the BioRad D-100 in our laboratory. It is a HPLC-based technology, similar to the BioRad VARIANT II that performs the hemoglobin variant analysis with the Beta-Thal program. Although HbA1c results are not based of any peaks after A0, any variants, if present, are marked in designated windows. A medical chart review is performed, and if the condition of hemoglobin variant is not mentioned, it is considered as “undiagnosed”, and vise versa.

Results

In the duration of 95 days, a total of 35684 HbA1c samples were analyzed. Of those, 491 samples (1.4%) were marked with hemoglobin variant present. The rest of the samples (98.6%, n=35193) had no hemoglobin variant marked. Of the 491 samples marked with variants, hemoglobin S accounted for 72.5% (n=356), hemoglobin C accounted for 19.1% (n=94), hemoglobin E accounted for 4.9% (n=24), and the rest were marked with various hemoglobinopathies (3.5%, n=17). In any category, vast majority of the patients were “undiagnosed”, ranging from 85% to 100%. Only 8% to 15% of the patients were aware of their hemoglobin variant. There is no difference of awareness in different age groups, either, given that hemoglobin variants are included in the Newborn Screen Program in most states. One patient, at age 55, with hemoglobin C disease (HbCC) was undiagnosed.

Conclusions

Although most patients who are carriers of hemoglobin variants can have a normal life and often go undiagnosed, it still poses risks in specific situations. Sickle cell trait patients are under risks at high-intensity physical activity. Carriers of hemoglobin C and hemoglobin E often have microcytosis but are misled as iron-deficiency anemia. Screening of hemoglobinopathies has become increasingly important in antenatal diagnosis and prevention of hemoglobin disorders.
A-178 Quality Improvement of Cystic Fibrosis-Related Diabetes Screening at a Large Pediatric Center
Authors: Chokkalla A , Tuley P , Kurtca M , Ona H , Ruiz F E , Devaraj S
Published: October 2024

Background

Cystic Fibrosis Foundation guidelines recommend annual diabetes screening by oral glucose tolerance test in pediatric patients with cystic fibrosis starting at the age of 10 years. Adherence to these guidelines proves to be challenging, and the nationwide screening rates are still considered suboptimal. The aim of this study is to assess and improve the screening rates at our large pediatric center.

Methods

A four-year retrospective audit of oral glucose tolerance test completion among pediatric patients with cystic fibrosis of age >10 years who have not yet been diagnosed with diabetes was conducted. A collaborative working group with the cystic fibrosis center was formed to identify the barriers to screening (Figure 1) and formulate a quality improvement plan, which was evaluated for a 9-month period post-intervention. The 3 key interventions included: 1) scheduling annual summer labs for all the eligible patients, 2) best practice advisory to guide providers with oral glucose tolerance test ordering, and 3) a streamlined process for oral glucose tolerance test appointments at the outpatient lab.

Results

The diabetes screening rates determined by oral glucose tolerance test completion at our center showed a gradual decline during the COVID-19 pandemic from 2019 to 2022. Following the implementation of the quality improvement plan during the summer of 2023, there was a marked increase in test ordering compliance by the provider as well as test completion by the patients. Notably, the fractional test completion rate rose from 45% during the pre-intervention phase (January-April, 2023) to 70% during the post-intervention phase (May-September, 2023).

Conclusions

Diabetes screening in pediatric patients with cystic fibrosis can be effectively improved by refining the practices related to patient experience, care coordination and laboratory testing strategies.
A-187 Program for Recognition of Patient Safety Cores in Diagnostic Services
Authors: Dias C , Marques F A , Morales A M , Canali D G , Teixeira M R , Mata L S , d. Melo G , Godoi C T , Quiarato P A , Maximo A C
Published: October 2024

Background

Health services are complex and challenging structures, including diagnostic services. Therefore, it is everyone's responsibility to ensure patient safety. The scope of this study is to present the challenges and opportunities in structuring the Quality Management and Patient Safety System (QM&PS) of a large diagnostic service that culminated in the first laboratory to receive the Patient Safety Core Recognition Seal (NSP), a program established by the partnership of QGA (Quality Global Assurance) and IBSP (Brazilian Institute for Patient Safety).

Methods

The restructuring of the QM&PS began in 2018 and since then, 6 NSPs have been established, distributed across 6 regions of the Brazil, with procedures, regulations, confidentiality agreements, and responsibilities clearly defined. The NSP are multidisciplinary and discuss 13 safety indicators monthly, namely: adverse events, contrast extravasation, falls and faints, reactions to medications, contrasts, and vaccines, immunization errors - vaccine application and exchange, loss of noble samples, technovigilance, and pharmacovigilance. With an engaged and participative Medical Board, the Patient Safety Committee was created in 2020, with the participation of top management and representatives of strategic leaderships to discuss cases of serious adverse events and deaths with systemic outputs and plans. In 2022, we conducted the first Patient Safety Perception Survey, from which we extracted insights to further develop the safety culture. Six working groups were organized focusing on interaction between company committees and programs, institutional communication for sharing information and Patient Safety initiatives in the organization, identification, prevention, and monitoring of incidents, risk management tool for analysis of clarity, application, and relevance of use as support for management and continuing education.

Results

The working groups involved 58 professionals and resulted in 65 implemented actions. Among the main results achieved, we can highlight the implementation of the rapid response flow to SAE (serious adverse event), Just Culture Policy, Disclosure Protocol, Second Victim Care Flow, Safety Rite in Customer Service Units, Patient Safety Trail, Institutional Communication, and the Patient Safety Core Recognition Seal.

Conclusions

Patient Quality and Safety must be part of the institutional pillar, composing the Strategic Objectives to be achieved. It is necessary to engage the organization and rely on the involvement of leadership and physicians. As a large service, our biggest challenges are demonstrating the effectiveness of implemented improvement actions in the face of identified risk modifications, defining strategies to reduce underreporting, and being able to involve all collaborators and operational teams in the safety culture.