ICSH guidance for internal quality control policy for blood cell counters.

This paper is a description of the ICSH guidance for internal quality control (IQC) policy for blood cell counters. It follows from and links to a separate ICSH review for such policies and practices. The ICSH has gathered information regarding the current state of practice through review of published guidance from regulatory bodies, a questionnaire to six major cell counter manufacturers and a survey issued to 191 diagnostic laboratories in four countries (China, the Republic of Ireland, Spain, and the United Kingdom) on their IQC practice and approach to the use of commercial IQC materials. This has revealed diversity both in guidance and in practice around the world. There is diversity in guidance from regulatory organizations in regard to IQC methods each recommends, clinical levels to use and frequency to run commercial controls, and finally recommended sources of commercial control materials. The diversity in practice among clinical laboratories spans the areas of IQC methods used, derivation of target values, and action limits used with commercial control materials, and frequency of running commercial controls materials. These findings and their implications for IQC Practice are addressed in this guidance document, which proposes a harmonized approach to address the issues faced by diagnostic laboratories.

ICSH review of internal quality control policy for blood cell counters.

INTRODUCTION: This paper is a report of an ICSH review of policies and practices for internal quality control (IQC) policy for haematology cell counters among regulatory bodies, cell counter manufacturers and diagnostic laboratories. It includes a discussion of the study findings and links to separate ICSH guidance for such policies and practices. The application of internal quality control (IQC) methods is an essential pre-requisite for all clinical laboratory testing including the blood count (Full Blood Count, FBC, or Complete Blood Count, CBC). METHODS: The ICSH has gathered information regarding the current state of practice through review of published guidance from regulatory bodies, a questionnaire to six major cell counter manufacturers (Abbott Diagnostics, Beckman Coulter, Horiba Medical Diagnostic Instruments & Systems, Mindray Medical International, Siemens Healthcare Diagnostics and Sysmex Corporation) and a survey issued to 191 diagnostic laboratories in four countries (China, Republic of Ireland, Spain and the United Kingdom) on their IQC practice and approach to use of commercial IQC materials. RESULTS: This has revealed diversity both in guidance and in practice around the world. There is diversity in guidance from regulatory organizations in regard to IQC methods each recommends, clinical levels to use and frequency to run commercial controls, and finally recommended sources of commercial controls. The diversity in practice among clinical laboratories spans the areas of IQC methods used, derivation of target values and action limits used with control materials, and frequency of running commercial controls materials. CONCLUSIONS: These findings and their implications for IQC Practice are discussed in this paper. They are used to inform a separate guidance document, which proposes a harmonized approach to address the issues faced by diagnostic laboratories.

Reference Interval Harmonization: Harnessing the Power of Big Data Analytics to Derive Common Reference Intervals across Populations and Testing Platforms.

BACKGROUND: Harmonization in laboratory medicine is essential for consistent and accurate clinical decision-making. There is significant and unwarranted variation in reference intervals (RIs) used by laboratories for assays with established analytical traceability. The Canadian Society of Clinical Chemists (CSCC) Working Group on Reference Interval Harmonization (hRI-WG) aims to establish harmonized RIs (hRIs) for laboratory tests and support implementation. METHODS: Harnessing the power of big data, laboratory results were collected across populations and testing platforms to derive common adult RIs for 16 biochemical markers. A novel comprehensive approach was established, including: (a) analysis of big data from community laboratories across Canada; (b) statistical evaluation of age, sex, and analytical differences; (c) derivation of hRIs using the refineR method; and (d) verification of proposed hRIs across 9 laboratories with different instrumentation using serum and plasma samples collected from healthy Canadian adults. RESULTS: Harmonized RIs were calculated for all assays using the refineR method, except free thyroxine. Derived hRIs met proposed verification criterion across 9 laboratories and 5 manufacturers for alkaline phosphatase, albumin (bromocresol green), chloride, lactate dehydrogenase, magnesium, phosphate, potassium (serum), and total protein (serum). Further investigation is needed for some analytes due to failure to meet verification criteria in one or more laboratories (albumin [bromocresol purple], calcium, total carbon dioxide, total bilirubin, and sodium) or concern regarding excessively wide hRIs (alanine aminotransferase, creatinine, and thyroid stimulating hormone). CONCLUSIONS: We report a novel data-driven approach for RI harmonization. Findings support feasibility of RI harmonization for several analytes; however, some presented challenges, highlighting limitations that need to be considered in harmonization and big data analytics.

Retrospective analysis of intra-patient laboratory variation demonstrates that the BD Vacutainer® Barricor™ blood collection tube reduces troponin variation.

OBJECTIVE: The BD Vacutainer® Barricor™ plasma blood collection tube uses a mechanical separator during centrifugation to separate plasma from the cellular elements of blood. Compared to use of plasma separator tubes (PST™) with gel, Barricor™ produces a cleaner sample with less residual cellular content. We sought to determine if Barricor™ reduces pre-analytical error compared to PST™. DESIGN & METHODS: We used a model previously published that utilizes serial differences between intra-patient measurements transformed into a Taylor series of variation vs time with the y-intercept equal to the sum of short-term analytic variation, preanalytic variation and biologic variation. The intra-patient variation of chloride, sodium, potassium, and troponin-T (hs-TnT) obtained from the Emergency Department of a large tertiary care center sampled with PST™ (May 2015-April 2018, n = 59,762 specimens) or Barricor™ (May 2018-May 2021, n = 61,512 specimens) was evaluated. All specimens were analyzed on either Roche Modular or Cobas® instruments. For each analyte, pairs of intra-patient results were tabulated and separated by 1 h intervals. The average between-pair variations were then regressed against time. We also determined the number of intra-patient outliers using the reference change value for each analyte. RESULTS: The Barricor™ hs-TnT y-intercept (-0.0132) was significantly lower than the PST™ intercept (0.9109; p = 0.022). This was also true for chloride (y-intercept = 1.0067 in Barricor™ and 1.3431 in PST™, p = 0.037). The percentage of hs-TnT outliers was significantly lower in Barricor™ (8.32 %) vs PST™ (12.2 %; p < 0.001). CONCLUSION: The analytical and biological variations are assumed to be steady over the study periods; we ascribe the difference in the y-intercept to the preanalytical effect of the Barricor™ tube reducing platelets and other cellular debris.

Transformation of Sequential Hospital and Outpatient Laboratory Data into Between-Day Reference Change Values.

BACKGROUND: Serial differences between intrapatient consecutive measurements can be transformed into Taylor series of variation vs time with the intersection at time = 0 (y0) equal to the total variation (analytical + biological + preanalytical). With small preanalytical variation, y0, expressed as a percentage of the mean, is equal to the variable component of the reference change value (RCV) calculation: (CVA2 + CVI2)1/2. METHODS: We determined the between-day RCV of patient data for 17 analytes and compared them to healthy participants' RCVs. We analyzed 653 consecutive days of Dartmouth-Hitchcock Roche Modular general chemistry data (4.2 million results: 60% inpatient, 40% outpatient). The serial patient values of 17 analytes were transformed into 95% 2-sided RCV (RCVAlternate), and 3 sets of RCVhealthy were calculated from 3 Roche Modular analyzers' quality control summaries and CVI derived from biological variation (BV) studies using healthy participants. RESULTS: The RCVAlternate values are similar to RCVhealthy derived from known components of variation. For sodium, chloride, bicarbonate calcium, magnesium, phosphate, alanine aminotransferase, albumin, and total protein, the RCVs are equivalent. As expected, increased variation was found for glucose, aspartate aminotransferase, creatinine, and potassium. Direct bilirubin and urea demonstrated lower variation. CONCLUSIONS: Our RCVAlternate values integrate known and unknown components of analytic, biologic, and preanalytic variation, and depict the variations observed by clinical teams that make medical decisions based on the test values. The RCVAlternate values are similar to the RCVhealthy values derived from known components of variation and suggest further studies to better understand the results being generated on actual patients tested in typical laboratory environments.

Average of Patient Deltas: Patient-Based Quality Control Utilizing the Mean Within-Patient Analyte Variation.

BACKGROUND: Because traditional QC is discontinuous, laboratories use additional strategies to detect systematic error. One strategy, the delta check, is best suited to detect large systematic error. The moving average (MA) monitors the mean patient analyte value but cannot equitably detect systematic error in skewed distributions. Our study combines delta check and MA to develop an average of deltas (AoD) strategy that monitors the mean delta of consecutive, intrapatient results. METHODS: Arrays of the differences (delta) between paired patient results collected within 20-28 h of each other were generated from historical data. AoD protocols were developed using a simulated annealing algorithm in MatLab (Mathworks) to select the number of patient delta values to average and truncation limits to eliminate large deltas. We simulated systematic error by adding bias to arrays for plasma albumin, alanine aminotransferase, alkaline phosphatase, amylase, aspartate aminotransferase, bicarbonate, bilirubin (total and direct), calcium, chloride, creatinine, lipase, sodium, phosphorus, potassium, total protein, and magnesium. The average number of deltas to detection (ANDED) was then calculated in response to induced systematic error. RESULTS: ANDED varied by combination of assay and AoD protocol. Errors in albumin, lipase, and total protein were detected with a mean of 6 delta pairs. The highest ANDED was calcium, with a positive 0.6-mg/dL shift detected with an ANDED of 75. However, a negative 0.6-mg/dL calcium shift was detected with an ANDED of 25. CONCLUSIONS: AoD detects systematic error with relatively few paired patient samples and is a patient-based QC technique that will enhance error detection.

Five-Year Two-Center Retrospective Comparison of Central Laboratory Glucose to GEM 4000 and ABL 800 Blood Glucose: Demonstrating the (In)adequacy of Blood Gas Glucose.

PURPOSE: To evaluate the glucose assays of two blood gas analyzers (BGAs) in intensive care unit (ICU) patients by comparing ICU BGA glucoses to central laboratory (CL) glucoses of almost simultaneously drawn specimens. METHODS: Data repositories provided five years of ICU BGA glucoses and contemporaneously drawn CL glucoses from a Calgary, Alberta ICU equipped with IL GEM 4000 and CL Roche Cobas 8000-C702, and an Edmonton, Alberta ICU equipped with Radiometer ABL 800 and CL Beckman-Coulter DxC. Blood glucose analyzer and CL glucose differences were evaluated if they were both drawn either within ±15 or ±5 minutes. Glucose differences were assessed graphically and quantitatively with simple run charts and the surveillance error grid (SEG) and quantitatively with the 2016 Food and Drug Administration guidance document, with ISO 15197 and SEG statistical summaries. As the GEM glucose exhibits diurnal variation, CL-arterial blood gas (ABG) differences were evaluated according to time of day. RESULTS: Compared to the GEM glucoses measured between 0200 and 0800, the run charts of (GEM-CL) glucose demonstrate significant outliers between 0800 and 0200 which are identified as moderate to severe clinical outliers by SEG analysis (P < .002 and P < .0005 for 5- and 15-minute intervals). Over the entire 24-hour period, the rates of moderate to severe glucose clinical outliers are 3.5/1000 (GEM) and 0.6/1000 glucoses (ABL), respectively, using the 15-minute interval (P < .0001). DISCUSSION: The GEM ABG glucose is associated with a higher frequency of moderate to severe glucose clinical outliers, especially between 0800 and 0200, increased CL testing and higher average patient glucoses.

Analytical Concordance of Diverse Point-of-Care and Central Laboratory Troponin I Assays.

BACKGROUND: Cardiac troponin I (cTnI) 99th percentile cutoffs, used in the diagnosis of acute myocardial infarction, are not standardized across cTnI assays. We compared 3 point-of-care (POC) and 1 central laboratory contemporary cTnI assays against the Abbott high-sensitivity (hs) cTnI to evaluate the analytical concordance and the feasibility of using a single cutoff value for all assays. METHODS: Fresh blood samples collected from 102 inpatients in the coronary care unit were measured on central laboratory instruments (Beckman Coulter DxI AccuTnI+3 TnI, Abbott Architect hs-TnI) and cTnI POC analyzers (Alere Triage Troponin I, Radiometer AQT90, Abbott i-STAT). Agreement and correlation between the contemporary cTnI assays and hs-cTnI assay were assessed using regression analysis. Proportional bias was assessed using Bland-Altman plots. Concordance between the contemporary cTnI and hs-cTnI assays was determined by diagnostic contingency tables at specific cutoffs. RESULTS: Most POC cTnI assays had excellent correlation with the Abbott hs-cTnI method (r 2 = 0.955-0.970) except for Alere Triage (r 2 = 0.617), while proportional bias is evident between all cTnI assays. Overall concordance between POC contemporary cTnI assays and hs-cTnI assay was 80% to 90% at their respective 99th percentile cutoffs. The concordance increased to 90% to 95% when a fixed cutoff of 0.03 to 0.05 ng/mL was used across the assays. CONCLUSIONS: This study demonstrates poor analytical concordance between cTnI assays at the 99th percentile and supports the notion of a single clinical decision limit for cTnI and consequently standardization of diagnostic protocols despite the analytical differences among these assays.

Barricor blood collection tubes are equivalent to PST for a variety of chemistry and immunoassay analytes except for lactate dehydrogenase.

INTRODUCTION: The BD Barricor tube uses a novel mechanical separator designed to eliminate gel artifacts, decrease cellular contamination, and improve stability. Here, we evaluated the Barricor tube as a possible replacement for PST using Beckman Coulter analyzers under both optimal, alternative, and suboptimal centrifugation conditions based on BD recommendations. METHODS: Paired PST and Barricor samples were collected from 4 local hospitals and processed based on site-specific preanalytical systems involving automated or manual centrifugation. Centrifugation conditions ranged from 1912 ×g for 10 min (suboptimal), 2060 g for 10 min (alternative), and 4000 ×g for 3 or 10 min (optimal). Tube volume (4.5 vs. 5.5 ml) was also assessed. Forty-three chemistry and immunochemistry analytes were measured on Beckman Coulter DxC and DxI analyzers. RESULTS: Using an automated preanlaytical system with suboptimal spin conditions, no bias between PST and Barricor was observed for all analytes tested except lactate dehydrogenase (LD). Further investigation revealed significant increase in LD when Barricor was spun for 10 min at 1912, 2060 and 4000 ×g, ranging from +7.4-19.4% vs. PST across the entire measurement interval (87-493 U/l). Smaller tube volume was also associated with higher LD. Differences in LD occurred despite no change in other hemolysis markers such as potassium, phosphate, and AST. CONCLUSIONS: LD is most sensitive to varying centrifugation conditions (time and speed) in Barricor tubes. We recommend that BD centrifugation protocols should be closely evaluated to determine if Barricor is equivalent to PST under local preanalytical configurations.

National Survey of Adult and Pediatric Reference Intervals in Clinical Laboratories across Canada: A Report of the CSCC Working Group on Reference Interval Harmonization.

OBJECTIVE: Reference intervals are widely used decision-making tools in laboratory medicine, serving as health-associated standards to interpret laboratory test results. Numerous studies have shown wide variation in reference intervals, even between laboratories using assays from the same manufacturer. Lack of consistency in either sample measurement or reference intervals across laboratories challenges the expectation of standardized patient care regardless of testing location. Here, we present data from a national survey conducted by the Canadian Society of Clinical Chemists (CSCC) Reference Interval Harmonization (hRI) Working Group that examines variation in laboratory reference sample measurements, as well as pediatric and adult reference intervals currently used in clinical practice across Canada. DESIGN AND METHODS: Data on reference intervals currently used by 37 laboratories were collected through a national survey to examine the variation in reference intervals for seven common laboratory tests. Additionally, 40 clinical laboratories participated in a baseline assessment by measuring six analytes in a reference sample. RESULTS: Of the seven analytes examined, alanine aminotransferase (ALT), alkaline phosphatase (ALP), and creatinine reference intervals were most variable. As expected, reference interval variation was more substantial in the pediatric population and varied between laboratories using the same instrumentation. Reference sample results differed between laboratories, particularly for ALT and free thyroxine (FT4). Reference interval variation was greater than test result variation for the majority of analytes. CONCLUSION: It is evident that there is a critical lack of harmonization in laboratory reference intervals, particularly for the pediatric population. Furthermore, the observed variation in reference intervals across instruments cannot be explained by the bias between the results obtained on instruments by different manufacturers.

The BD Barricor blood collection tube is an acceptable and robust alternative to the PST for use with the Beckman AccuTnI+3 assay.

OBJECTIVES: BD Canada recently released a blood collection tube with a novel mechanical separator called the Barricor. We evaluated this tube as an alternate sample type for cardiac troponin I (cTnI) testing using the Beckman Coulter AccuTnI+3 assay. DESIGN AND METHODS: 3014 paired patient specimens (Barricor, plasma separator tube or PST) were obtained from the emergency departments and cardiac care units of nine hospitals in and around Edmonton, Alberta. After centrifugation, each plasma sample was analyzed for cTnI using the Beckman Coulter AccuTnI+3 assay. In addition, selected samples were analyzed multiple times within a single run or over 4-5days to generate imprecision data for the assay. RESULTS: Repeatability and within-laboratory studies revealed an imprecision of <10% at concentrations above 0.025µg/L for the Barricor as well as BD's traditional PST. Paired patient sample comparisons over the full range of the assay yielded linear regression slopes ranging from 0.956 to 1.011 and Pearson correlation coefficients ranging from 0.993 to 0.999. At a lower range of results closer to the manufacturer's 99th percentile cutoffs correlation was slightly worse, but still acceptable, with linear regression slopes ranging from 0.967 to 1.211 and Pearson correlation coefficients ranging from 0.983 to 0.987. Notably, at these lower concentrations the agreement between individual PST and Barricor results worsened with decreasing cTnI concentration. Differences between pairs of results became particularly large (-50 to +400%) at PST cTnI concentrations ≤0.015µg/L. Closer inspection of the data around the 0.02 and 0.04µg/L 99th percentile cutoffs revealed a number of discordances between PST and Barricor results, with at least some of these attributable to false elevations in the PST results. CONCLUSIONS: Together, our results suggest that the Barricor blood collection tube is good alternative to the traditional PST for cTnI testing using the AccuTnI+3 assay. The Barricor appears to minimize spurious, nonreproducible, and false elevations in cTnI results for a subset of patients but additional studies are needed to determine if it reduces overall false elevations. cTnI results below 0.04µg/L may still be of questionable accuracy even with the use of this new tube.

Impaired clinical utility of sequential patient GEM blood gas measurements associated with calibration schedule.

BACKGROUND: Within- and/or between-instrument variation may falsely indicate patient trends or obscure real trends. We employ a methodology that transforms sequential intra-patient results into estimates of biologic and analytic variation. We previously derived realistic biologic variation (sb) of blood gas (BG) and hematology analytes. We extend this methodology to derive the imprecision of two GEM 4000 BG analyzers. METHODS: A laboratory data repository provided arterial BG, electrolyte and metabolite results generated by two GEM 4000s on ICU patients in 2012-2013. We tabulated consecutive pairs of intra-patient results separated by increasing time interval between consecutive tests. The average between pair variations were regressed against time with the y-intercept representing the sum of the biologic variation and short term analytic variation: yo2=sb2+sa2. Using an equivalent equation for the Radiometer ABL, the imprecision of the two GEMs was calculated: saGEM=(yoGEM2-yoABL2+saABL2)1/2. This analysis was performed for nearly all measurements, regardless of time as well for values obtained over two 12h mutually exclusive periods, starting either at 2am or 2pm. RESULTS: Regression graphs were derived from 1800 patients' blood gas results with least 10,000 data pairs grouped into 2h intervals. The calculated saGEM exceed the directly measured saABL with many GEM sigma ratios of biologic variation/analytic variation being close to unity. All of the afternoon saGEM exceeded their morning counterparts with pH, pCO2, K and bicarbonate being statistically significant. CONCLUSION: For many analytes, the average analytical variation of tandem GEMs approximates the biologic variation, indicating impaired clinical usefulness of tandem sequential measurements. A significant component of this variation is due to increased variation of the GEMs between 2pm and 2am.

Requirements for Successful Adoption of a Glucose Measurement System Into a Hospital POC Program.

Widespread and successful implementation of any glucose measurement system in a hospital point-of-care (POC) program requires a number of features in addition to accurate and reliable analytical performance. Such features include, but are not limited to, a system's glucose-hematocrit dependence, durability, information technology capabilities, and battery capacity and battery life. While the study of Ottiger et al in this issue supports the analytical accuracy and reliability of Bayer's CONTOUR XT® blood glucose monitoring system, the suitability of other features of this system for a hospital POC program remains to be established.

The Glucose Measurement Industry and Hemoglobin A1c: An Opportunity for Creative Destruction.

The MyStar Extra self-monitoring blood glucose (SMBG) system provides moving estimates of the patient's hemoglobin A1c (HbA1c). There is a treasure trove of highly accurate glucose data available from highly accurate SMBG, CGM and FGM along with highly accurate HPLC HbA1c. If Nathan's criteria are used to select subjects whose glucoses can be correlated to the HbA1c, then algorithms can be developed for robustly transforming glucose into HbA1c. These algorithms can then be implemented in any SMBG or with the CGM and FGM software. This calculated HbA1c would even be accurate with Nathan's excluded population thus reducing the use of fructosamine and glycated protein. Finally, the developer of these new algorithms is advised to use a specific approach for testing her algorithm.

Limiting the testing of AST: a diagnostically nonspecific enzyme.

OBJECTIVES: Annually, millions of pairs of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) tests are ordered. These enzymes are highly correlated, and ALT is far more specific diagnostically than AST. To reduce AST testing, we suggest measuring AST only when ALT exceeds a predetermined limit. METHODS: We derived the proportions of elevated ASTs that would not be measured based on 15 months of paired inpatient and outpatient ALT and AST data. RESULTS: For inpatients, a 35 U/L ALT limit for initiating AST testing would reduce AST testing by 51%, missing only 3% and 7.5% of ASTs exceeding 50 U/L and 35 U/L, respectively. In outpatients, AST testing can be reduced by more than 65%, with fewer missed elevated ASTs (0.5% and 2% of the ASTs exceeding 50 U/L and 35 U/L, respectively). CONCLUSIONS: Conservatively, $100 million could be saved annually in the US health care budget by selectively limiting AST testing in just the US outpatient environment.