External Quality Control Program Research Articles

Analysis of Influencing Factors on Participation Compliance, Slide Preparations Quality and Microscopic Reading Quality of Acid-Resistant Bacteria Slides at Ciamis Regency

Background & Objective: To find out the factors that influence participation compliance, slide preparations quality and microscopic reading quality in the External Quality Control program of Microscopic Cross Tests Acid Resistant Bacteria in Ciamis Regency in 2022-2023 Method: The research design is cross-sectional, population and total sampling of 27 respondents. Data analysis is Chi-Square with Fisher's Exact Test Result: Research data shows that the sig-p value of the period of employment variable for participation is 0.057, for the quality of slide preparations per quarter 0.695, 0.447, 0.550, 0.305, and the microscopic reading quality 0.036. The sig-p value of the training variable on participation is 0.226, on the quality of slide preparations per quarter 0.043, 0.388, 0.212, 0.219, and the microscopic reading quality 0.298. The sig-p value of the workload variable for participation is 0.204, for the quality of slide preparations per quarter 0.502, 0.091, 0.332, 0.502, and for the microscopic reading quality 0.050. The sig-p value of the microscope condition variable for participation is 0.005, for the quality of slide preparations per quarter 0.063, 0.106, 0.149, 0.323, and for the microscopic reading quality 0.036. The sig-p value of the incentive variable for participation is 0.001, for the quality of slide preparations per quarter 0.528, 0.302, 0.586, 0.189, for the microscopic reading quality of 0.013 Conclusion: there is a relationship between microscope conditions and incentive on participation compliance, there is a relationship between training on the slide preparations quality in the 4th quarter of 2022, there is a relationship between period of employment, workload, microscope conditions and incentive on microscopic reading quality

Ten-year results of an international external quality control programme for measurement of anti-tuberculosis drug concentrations.

Participation in an external (interlaboratory) quality control (QC) programme is an essential part of quality assurance as it provides laboratories with valuable insights into their analytical performance. We describe the 10 year results of an international QC programme for the measurement of anti-tuberculosis (TB) drugs. Each year, two rounds were organized in which serum (or plasma) samples, spiked with known concentrations of anti-TB drugs, were provided to participating laboratories for analysis. Reported measurements within 80%-120% of weighed-in concentrations were considered accurate. Mixed model linear regression was performed to assess the effect of the measured drug, concentration level, analytical technique and performing laboratory on the absolute inaccuracy. By 2022, 31 laboratories had participated in the QC programme and 13 anti-TB drugs and metabolites were included. In total 1407 measurements were reported. First-line TB drugs (isoniazid, rifampicin, pyrazinamide and ethambutol) represented 58% of all measurements. Overall, 83.2% of 1407 measurements were accurate, and the median absolute inaccuracy was 7.3% (IQR, 3.3%-15.1%). The absolute inaccuracy was related to the measured anti-TB drug and to the performing laboratory, but not to the concentration level or to the analytical technique used. The median absolute inaccuracies of rifampicin and isoniazid were relatively high (10.2% and 10.9%, respectively). The 10 year results of this external QC programme illustrate the need for continuous external QC for the measurement of anti-TB drugs for research and patient care purposes, because one in six measurements was inaccurate. Participation in the programme alerts laboratories to previously undetected analytical problems.

The Credibility of Microbiological Diagnostics in MedicalDiagnostic Laboratories Based on the Results of POLMICROExternal Laboratory Quality Control POLMICRO 2023

<br>Introduction: The article describes results of the 30th edition of the Polish National ExternalQuality Assessment Scheme in Microbiology POLMICRO organised by the Centre of QualityControl in Microbiology (CQCM).</br><br>Aim: The evaluation of the reliability of the microbiological diagnostics performed bymedical diagnostic laboratories in Poland.</br><br>Materials and methods: The organiser analysed the results achieved by participants ofprogrammes conducted by CQCM in 2023 concerning important stages of microbiologicaldiagnostics such the as microscopic analysis of Gram-stained slides, identificationof etiological factors of infections, assessment of antimicrobial susceptibility ofmicroorganisms and the detection of antimicrobial resistance mechanisms.</br><br>Results: A positive annual assessment in the POLMICRO 2023 programme was achieved by98.33%, so 413 of 420 participants of the POLMICRO 2023 programme, and all laboratories participatingin the POLMICRO/SSE (n = 132) and POLMICRO/MIKOLOGIA (n = 139) programmes.</br><br>Conclusions: The results of external quality control programmes confirm the reliability oftesting and the high level of microbiological diagnostics performed by medical diagnosticlaboratories in Poland.</br>

Clinic-based evaluation of the dual Xpert CT/NG assay on the GeneXpert System for screening for extragenital chlamydial and gonococcal infections amongst men who have sex with men

BackgroundChlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) infections have increased globally. Asymptomatic infections represent a significant risk of long-term complications. Men who have sex with men (MSM) are disproportionally affected, underscoring the need to offer screening programmes to this population. CT/NG Point of Care Testing (POCT) constitutes a strategic tool to improve the continuum of STI care, however extensive real-life evaluations amongst at risk populations are lacking. The aim of this study is to estimate the GeneXpert CT/NG assay performance and usability for CT and NG at genital and extragenital sites for screening amongst MSM.MethodsThis study was a multi-site sexual health clinic-based evaluation (Italy, Malta and Peru) with consecutive enrolment. A first void urine sample (divided in two aliquots), two oropharyngeal and two anorectal swabs were collected for each study participant. One specimen set (one for each anatomical site) was tested with the dual index test (Cepheid) at the clinics by the healthcare staff, the other set with FDA/CE approved Nucleic Acid Amplification Tests (NAATs) at the laboratory. Clinical sites and reference laboratories participated in an internal and external quality control programme. Sensitivity, specificity, positive and negative likelihood ratios, positive and negative predictive values for each anatomical site were estimated using a meta-analytic approach.ResultsOne thousand seven hundred two MSM were recruited across all clinical sites for a total of 5049 biological specimens. NG and CT were respectively detected in 274 and 287 of samples. Overall, the NG POCT sensitivity and specificity was 91.43% and 99.75% in urine (LR + 372.80, LR- 0.09), 89.68% and 99.55% in rectal specimens (LR + 197.30, LR- 0.10) and 75.87% and 98.77% at the pharynx respectively (LR + 61.94, LR- 0.24). The CT component of the POCT sensitivity was 84.82% and specificity 99.63% in urine (LR + 228.68, LR- 0.15), 78.07% and 99.19% respectively on rectal site (LR + 96.23, LR-0.22), 67.79% and 99.88% respectively at pharyngeal site (LR + 554.89, LR- 0.32). 95.95% of MSM reported to be willing to wait for POCT results and no provider reported difficulties in terms of performance or interpretation of the results of the Xpert CT/NG.ConclusionRapid turnaround time, ease of use and high acceptability make the Xpert CT/NG testing system a strategic tool for increasing testing frequency, reaching those not yet tested and offering the possibility of immediate treatment if needed. The assay showed good negative likelihood ratios and confirms its use to rule out CT/NG infections. Sensitivity varied across sites and pathogens. Periodic staff training at the testing sites should be mandatory.

External quality control program in infectious diseases screening at laboratories and blood banks in Latin America: an analysis of the past 5 years.

To evaluate the screening of blood samples for infectious disease markers at laboratories and blood banks in Latin America per the findings of an External Quality Assessment Program (EQAP). This qualitative analysis used data from the EQAP coordinated by the Fundação Pro Sangue Hemocentro de São Paulo with the support of the Pan American Health Organization to assess the performance of blood screening for infectious diseases from 2014 to 2018 in Latin America. Each participating laboratory or blood bank received an identical blind panel with 24 blood samples with variable reactivity for all the screening parameters. Panels were processed at each participating facility and results were returned to the Fundação Pro Sangue Hemocentro de São Paulo for individual and joint analyses. Two types of discrepant results were potential failures: false positive results (FPRs) and false nonreactive results (FNRRs). A total of 23 136 samples were evaluated. Global rates of FPR, FNRR, and concordant results were 0.3%, 1.0% and 98.7%, respectively. Seven FNRRs were found for HBsAg (1.0%), 12 for syphilis (2.6%), and 21 for Chagas disease (2.9%). No FNRRs were found for the HIV, HCV, and HTLV viruses. The average accuracy of all the laboratories and blood banks participating in the EQAP during the study period was 99.5% (standard deviation, 0.5%). The findings of this qualitative analysis are positive for blood safety in Latin America, with an average accuracy of 99.5% among the participating laboratories and blood banks. This report reflects an important improvement in blood bank serological screening EQAP-PAHO report since the 2003.

Matrix-matched calibrators are necessary for robust and high-quality dried blood spots lead screening assays by inductively coupled plasma-mass spectrometry

Background and aimsReliable lead screening methods are necessary to support early identification of lead exposure in children. Sample collection using dried blood spots (DBS) offers advantages compared to traditional venipuncture and capillary collection. Here, we describe and compare three lead DBS inductively coupled plasma-mass spectrometry (ICP-MS) methods for lead screening. Materials and methodsLead was extracted from Whatman 903 protein saver cards punches and analyzed by ICP-MS across three independent clinical laboratories. Each laboratory evaluated the performance of aqueous and matrix-matched DBS calibrators using external quality control samples (WI State of Laboratory of Hygiene Program). Leftover patient samples (n = 39) were used for an interlaboratory comparison of lead DBS. Lead DBS results were compared to whole blood methods. ResultsThe DBS ICP-MS methods using matrix-matched DBS calibrators had superior performance to the aqueous calibrations. There was a strong correlation between lead measured in DBS (matrix-matched) and whole blood for the three methods evaluated. ConclusionLead can be measured accurately by ICP-MS in DBS samples when matrix-matched calibrators are used. External quality control programs are valuable to assess the performance of DBS methods. DBS lead ICP-MS methods are a robust analytical option for lead screening even though the limitations of DBS are well recognized.

A-062 Development of Quality Control Indicators in the Pre-analytical Phase and a Survey on the Current Status of Clinical Laboratories in Korea

Abstract Background Pre-analytical errors, which account for 50%–75% of all laboratory errors, include various processes handled by various occupations such as prescription, data input, sample collection, sample transport, and sample reception, making it difficult to present standardized quality indicators. Internationally, efforts are being made to present pre-analytical quality indicators and standardize/harmonize them, but in Korea, the status of pre-analytical errors in laboratories has not been identified. Therefore, this study attempted to contribute to improving the quality of domestic clinical laboratories by presenting pre-analytical quality indicators that can be easily applied and effectively managed in domestic clinical laboratories. Methods Domestic and foreign programs for pre-analytical quality indicators were investigated, and a pre-analytical quality indicator survey was conducted for domestic clinical laboratories. The survey results were analyzed to identify the current status and problems of quality management in the pre-analytical stage of the domestic clinical laboratories, and the appropriateness of the pre-analytical quality indicators in the application of the domestic clinical laboratories was reviewed. Results We prepared 22 pre-analytical quality indicators by referring to the “Quality indicators of the Pre-analytical phase” presented by the International Federation of Clinical Chemistry Working Group (2017) and “Development of an external quality control program in the pre-analytical stage for reliable laboratory implementation” presented by the Korean Association of External Quality Assessment (2021). A survey was conducted on 90 institutions nationwide, including tertiary hospitals, secondary hospitals, and professional consignment laboratories. As a result of the survey, 42 institutions (47%) responded, of which 64% were tertiary hospitals, 22% were secondary hospitals, and 14% were professional consignment laboratories. Among the quality indicators, the rate of complete blood counts accompanied by blood clots (93%) and the insufficient sample volume rate (90%) showed the highest response rate. On the other hand, the sample recollection rate (17% in the out of laboratory, 21% in the laboratory), the ratio of samples carried at nonconforming temperatures (24%), and the ratio of delayed samples (24%) showed the lowest response rate, respectively. The median value of the clinical laboratory error rate for all quality indicators of pre-analytical phase was generally very good with a sigma value of 5 or higher. Conclusion In this study, 22 pre-analytical quality indicators applicable to domestic clinical laboratories were prepared. Using this, the status of quality management in the pre-analytical phase of the domestic clinical laboratory was identified, and the appropriateness of quality indicators was reviewed. This is an important cornerstone for the standardization of quality management in the pre-analytical phase, and it is expected that it will greatly contribute to improving the quality of clinical laboratories in Korea by establishing a pre-analytical quality management system in the future.

EAA/EMQN best practice guidelines for molecular diagnosis of Y-chromosomal microdeletions: State of the art 2023.

Testing for AZoospermia Factor (AZF) deletions of the Y chromosome is a key component of the diagnostic workup of azoospermic and severely oligozoospermic men. This revision of the 2013 European Academy of Andrology (EAA) and EMQN CIC (previously known as the European Molecular Genetics Quality Network) laboratory guidelines summarizes recent clinically relevant advances and provides an update on the results of the external quality assessment program jointly offered by both organizations. A basic multiplex PCR reaction followed by a deletion extension analysis remains the gold-standard methodology to detect and correctly interpret AZF deletions. Recent data have led to an update of the sY84 reverse primer sequence, as well as to a refinement of what were previously considered as interchangeable border markers for AZFa and AZFb deletion breakpoints. More specifically, sY83 and sY143 are nolonger recommended for the deletion extension analysis, leaving sY1064 and sY1192, respectively, as first-choice markers. Despite the transition, currently underway in several countries, toward a diagnosis based on certified kits, it should be noted that many of these commercial products are not recommended due to an unnecessarily high number of tested markers, and none of those currently available are, to the best of our knowledge, in accordance with the new first-choice markers for the deletion extension analysis. The gr/gr partial AZFc deletion remains a population-specific risk factor for impaired sperm production and a predisposing factor for testicular germ cell tumors. Testing for this deletion type is, as before, left at the discretion of the diagnostic labs and referring clinicians. Annual participation in an external quality control program is strongly encouraged, as the 22-year experience of the EMQN/EAA scheme clearly demonstrates a steep decline in diagnostic errors and an improvement in reporting practice.

The quality of microbiological diagnostics in Poland based onthe results of external laboratory quality control programmes

<br><b>Introduction:</b> The article describes the results of the 29th edition of the Polish NationalExternal Quality Assessment Scheme in Microbiology POLMICRO created by the Centre ofQuality Control in Microbiology (CQCM).</br><br><b>Aim:</b> The aim of the scheme is the evaluation of the reliability of the results of microbiologicaltests performed by Polish medical laboratories, which includes e.g. identification andassessment of antimicrobial susceptibility of bacterial etiological agents of infections,together with the detection of antimicrobial resistance mechanisms.</br><br><b>Materials and methods:</b> The assessment of external quality control programmes organised byCQCM is based on the analysis of the results of participants of the POLMICRO, POLMICRO/MIKOLOGIA and POLMICRO/SSE programmes in a full annual cycle.</br><br><b>Results:</b> In 2022, 96.00% of POLMICRO participants, as well as 99.22% of laboratories participatingin POLMICRO/SSE and all participants of POLMICRO/MIKOLOGIA, obtained positiveresults.</br><br><b>Conclusions:</b> The results of external quality control programmes conducted by CQCM showthat medical laboratories in Poland have been maintaining a high level of microbiologicaldiagnostics.</br>

Fifth Generation Quality Control of Immunoassay Parameters by Six Sigma Value.

To determine the sigma value of immunoassay parameters which are within the 2Z score on external quality control (EQC). A cross-sectional study. Place and Duration of the Study: Department of Chemical Pathology and Endocrinology (AFIP),non-ability ity consecutive from June to November 2022. Ten immunoassay parameters were selected on the basis of the internal quality control (IQC) and external quality control (EQC) programs. The Clinical Laboratory Improvement Amendments (CLIA) is used for Total Allowable Error (TEa). Sigma value was calculated from coefficient of variation (CV) and bias, which was determined by IQC and EQC, observed for 6 subsequent months. If the sigma values are ≥6, between 3 and 5, and <3, they are classified as good, acceptable or unacceptable, respectively. T4, prolactin, Vitamin B12 at >3 óat IQC level 1. TSH, T3, T4, and Vitamin B12 showed that on level 2 IQC at >3 óT3, Vitamin D at 4-5 ó level, Prolactin, FSH, and LH at 6 ó level in level 2 IQC. In the EQC program, the sigma level calculated for ten assays found that almost all parameters were at sigma >3 ó level except TSH which was at 5.8 ϭ level during June to August 2022. From September to November 2022, all parameters at >3 ó level except for TSH, growth hormone, FSH, LH, and Vitamin b12 which showed at 4.4 ó level. Most of the immunoassay parameters show good performance in the EQC program and at both levels of IQC level with sigma value 4-5. Bias, Six sigma, Key performance indicators, External quality control.

Assessment of the performance of Romanian Public Health Units in identifying and conducting antimicrobial susceptibility testing for healthcareassociated infection-causing pathogens

Detecting health-associated infectious (HAIs) is crucial as HAIs can pose a significant threat to patient safety, particularly among vulnerable populations such as the elderly, immunocompromised individuals, and those undergoing invasive medical procedures. Also, it is important to monitor the ef-fectiveness of antibiotics and identify the resistance patterns of microorganisms to ensure appropriate treatment options. Bacteriology external quality control programs provide a means for laboratories to assess their performance and identify areas that require improvement. By participating in such pro-grams, laboratories can evaluate their procedures, equipment, and personnel to ensure that they are producing reliable and accurate results because the External Quality Control (EQC) programs are carried out by medical laboratories using identical procedures as those used for handling patient samples. Our investigation centered on the performance of Romanian Public Health Units in identifying bacteria and conducting antimicrobial susceptibility testing in Bacteriology EQC programs associated with some of the most prevalent HAIs, such as blood infections, respiratory infections, and postoperative infections.

Harmonization of indirect reference intervals calculation by the Bhattacharya method.

The aim of this study was to harmonize the criteria for the Bhattacharya indirect method Microsoft Excel Spreadsheet for reference intervals calculation to reduce between-user variability and use these criteria to calculate and evaluate reference intervals for eight analytes in two different years. Anonymized laboratory test results from outpatients were extracted from January 1st 2018 to December 31st 2019. To assure data quality, we examined the monthly results from an external quality control program. Reference intervals were determined by the Bhattacharya method with the St Vincent's hospital Spreadsheet firstly using original criteria and then using additional harmonized criteria defined in this study. Consensus reference intervals using the additional harmonized criteria were calculated as the mean of four users' lower and upper reference interval results. To further test the operation criteria and robustness of the obtained reference intervals, an external user validated the Spreadsheet procedure. The extracted test results for all selected laboratory tests fulfilled the quality criteria and were included in the present study. Differences between users in calculated reference intervals were frequent when using the Spreadsheet. Therefore, additional criteria for the Spreadsheet were proposed and applied by independent users, such as: to set central bin as the mean of all the data, bin size as small as possible, at least three consecutive bins and a high proportion of bins within the curve. The proposed criteria contributed to the harmonization of reference interval calculation between users of the Bhattacharya indirect method Spreadsheet.

Antibiotic Susceptibility Testing (AST) Practices in India: Need for a National AST Forum.

Background Accurate interpretation of antibiotic susceptibility testing (AST) is one of the most crucial functions of the microbiology laboratory. However, its performance depends on a number of critical factors. We conducted a status survey to understand the existing practices in Indian laboratories that have a potential to influence performance of AST. Method We developed a 22-point online survey questionnaire on information about respondent's specifications, use of AST consumables, existing quality control protocols, and matters of contention in AST practices, and sent it by Google forms to 362 clinical microbiologists (holding MD or DNB certification). Participation was voluntary. Results were analyzed using descriptive statistics. Results Among 362, a total of 103 returned the questionnaire. The first 100 responses that were complete (all 22 questions answered) were analyzed. Respondents were from medical colleges (61%), private hospitals (26%), and stand-alone laboratories (13%). Analysis revealed that the Clinical & Laboratory Standards Institute (CLSI) guidelines were followed by all. Overall, 54% used disc diffusion as the primary method for performing AST. For the internal quality control testing of media and AST, 24% and 16% had adequate testing components and frequency, respectively. For performing AST of colistin, broth microdilution was used by 19%. Also, 86% participated in external quality control programs, and 54% respondents were dissatisfied or unsure about the development of competencies in AST methodology during their postgraduate training. Conclusion This survey reveals that potential gaps exist in the performance parameters and internal quality control of AST. There is an urgent need for harmonization in AST performance and postgraduate training in clinical microbiology in India.

External quality control in laboratory medicine. Progresses and future.

An external quality control program distributes same control samples to various laboratories and evaluates results obtained with a common criterion. The aim of this work is to summarize the evolution of various types of external programs, to point out the progresses ant to preclude practical consequences of the participant laboratories. The material consists on a brief revision of the different types of external programs that have been used for the last forty years. The method is the critical analysis of the strong and weak points of each program model, from the light of our experience.External quality assessment (EQA) programs were initiated at half the XX century, evidencing big discrepancies among laboratory results. EQA were developed in various countries and some mechanisms to harmonize them were proposed: to establish common performance specifications derived from biological variation, to use EQS as educational tool. Since the 2000 important advances were seen: to focus EQA to assure the adequate clinical use of laboratory tests, to use commutable controls, to harmonize the different EQA models, to promote a forum for co-operation and exchange of knowledge on quality-related matters for EQA organizers. To participate in an EQA with commutable-reference method assigned values controls allows to know the real inaccuracy of results and their impact on patient' samples. To participate in a EQA with non commutable controls allows to know whether the individual laboratory performance agrees with that from other laboratories using same analytical method.

Results From a Proficiency Testing Pilot for Immunosuppressant Microsampling Assays.

Therapeutic drug monitoring (TDM) of immunosuppressive drugs is important for the prevention of allograft rejection in transplant patients. Several hospitals offer a microsampling service that provides patients the opportunity to sample a drop of blood from a fingerprick at home that can then be sent to the laboratory by mail. The aim of this study was to pilot an external quality control program. Fourteen laboratories from 7 countries participated (fully or partly) in 3 rounds of proficiency testing for the immunosuppressants tacrolimus, ciclosporin, everolimus, sirolimus, and mycophenolic acid. The microsampling devices included the following: Whatman 903 and DMPK-C, HemaXis, Mitra, and Capitainer-B. All assays were based on liquid chromatography with tandem mass spectrometry. In round 2, microsamples as well as liquid whole blood samples were sent, and 1 of these samples was a patient sample. Imprecision CV% values for the tacrolimus microsamples reported by individual laboratories ranged from 13.2% to 18.2%, 11.7%-16.3%, and 12.2%-18.6% for rounds 1, 2, and 3, respectively. For liquid whole blood (round 2), the imprecision CV% values ranged from 3.9%-4.9%. For the other immunosuppressants, the results were similar. A great variety in analytical procedures was observed, especially the extraction method. For the patient sample, the microsample results led to different clinical decisions compared with that of the whole blood sample. Immunosuppressant microsampling methods show great interlaboratory variation compared with whole blood methods. This variation can influence clinical decision-making. Thus, harmonization and standardization are needed. Proficiency testing should be performed regularly for laboratories that use immunosuppressant microsampling techniques in patient care.

Effect of season, genetic line and temperature during transport on sperm motility of commercial insemination doses of pooled boar semen: A retrospective study.

Commercial pooled semen from boars of maternal and terminal genetic lines was analysed over two consecutive years, as part of an external quality control program. Semen doses were prepared for two total sperm counts (2.0 × 109 /75 ml [n=578] and 1.5 × 109 /34 ml [n=643]). Semen doses were evaluated for motility, morphology and bacterial contamination at days 1 and 4 after collection. Additionally, temperature during transport was tracked and correlated to sperm motility. Sperm motility and morphology were significantly affected by genetic line, but not by total sperm per doses. Total motility (TM) was affected (p < .05) by season. Progressive motility (PM) was lower (p < .05) in some winter months, and higher (p < .05) in summer for maternal-line doses. Normal sperm morphology was higher (p < .05) in some months of late fall and winter, with an apparent decrease in some summer months. In general, motility was significantly higher on day 1 than on day 4, and was also affected (p < .05) by temperature, especially for terminal-line doses. Semen doses exposed to >15°C had higher (p < .05) TM, VSL (straight-line velocity) and linearity than doses exposed to <15°C. Progressive motility and straightness increased (p < .05) proportionally with temperature. At day 1, motility, average path velocity (VAP) and VSL increased (p < .05) with temperature, and was higher than the corresponding group at day 4, except for VSL. Sperm motility and kinematics at day 4 were similar between temperature groups, except for doses exposed to >20°C that had higher (p < .05) VAP and VSL than doses exposed to <12°C. Microbiology evaluation revealed that less than 8.2% (15/184) of the semen doses had bacterial growth. Sphingobacterium lactis was the most prevalent bacterium identified. In conclusion, genetic line strongly influences sperm motility, while total sperm count does not have an impact. Transient reduced temperatures during shipping do not impair sperm quality after 4 days of storage. Seasonal and temperature effects on sperm motility of pooled boar semen are not overcome by pre-production ejaculate selection and semen pooling.

External quality control of microbiological testing in Polish medical diagnostic laboratories in 2020

&lt;br&gt;&lt;b&gt;Introduction:&lt;/b&gt;The paper presents the course of external quality control programmes organised in 2020 by the Centre of Quality Control in Microbiology (CQCM), including the 27&lt;sup&gt;th&lt;/sup&gt; edition of the Polish National External Quality Assessment Scheme in Microbiology POLMICRO.&lt;/br&gt; &lt;br&gt;&lt;b&gt;Aim:&lt;/b&gt;The assessment of the microbiological diagnostics’ quality carried out by Polish laboratories based on results of the POLMICRO, POLMICRO/MIKOLOGIA (MYCOLOGY), and POLMICRO/SSE programmes.&lt;/br&gt; &lt;br&gt;&lt;b&gt;Material and methods:&lt;/b&gt;The participants’ responses from all rounds conducted in 2020 were analysed, i.e. five rounds of the POLMICRO programme, two rounds of the POLMICRO /MIKOLOGIA programme, and one round of POLMICRO/SSE.&lt;/br&gt; &lt;br&gt;&lt;b&gt;Results:&lt;/b&gt;Positive evaluation in all rounds in the annual cycle were obtained by 90,1% of POLMICRO, 92,5% POLMICRO/MIKOLOGIA, and 100% POLMICRO/SSE participants.&lt;/br&gt; &lt;br&gt;&lt;b&gt;Conclusions:&lt;/b&gt; Laboratories participating in the CQCM programmes demonstrated skills in the identification, antimicrobial susceptibility testing, and detection of resistance mechanisms of pathogenic microorganisms. The area of internal quality control of antimicrobial susceptibility testing requires improvement, especially the selection of reference strains.&lt;/br&gt;

Generation of a single-tube quality control material for hemoglobin and DNA analyses of hemoglobinopathies.

Hemoglobinopathies are major public health problems worldwide. Accurate laboratory diagnosis of the carrier is essential, which includes initial screening, Hb analysis, and DNA analysis. For the first time, we have developed a single-tube quality control (QC) sample for these laboratory tests. The QC sample was made from a lyophilized mixture of the stabilized hemolysate with carbon monoxide saturation and the white blood cells of known thalassemia mutations. Homogeneity and stability were examined by Hb and DNA analyses on day 0 and every month for 12months, at room temperature, 4°C, and -20°C. A preliminary proficiency testing (PT) program for hemoglobinopathies using this single QC material was developed. Hemoglobin (Hb) and DNA analyses of a single-tube QC sample demonstrated satisfactory results of Hb analysis for at least five months and DNA analysis for at least one year of storage at -20°C. The results obtained from a preliminary PT program on five expert laboratories confirmed that a single tube QC sample prepared could be used as a PT item with various Hb and DNA analyses methods. A single lyophilized control sample has been generated for use in hemoglobinopathies' internal and external quality control program. Unlike other available control materials, which are used for individual testing, a single-tube QC sample generated can be used to control the pre-analytical and analytical processes of both Hb and DNA analyses and is suitable for use in the PT program of hemoglobinopathies.

O-149 Evaluation of structural problems in the application of strict criteria for sperm morphology assessment

Abstract Study question Which criteria, described by WHO 2010, cause most problems during sperm morphology assessment and lead to outcome variation in a national external quality control program? Summary answer Assessment of head ovality, regularly contoured head, regularly contoured midpiece and alignment of major axis of midpiece and head lead to the most variation. What is known already Morphology assessment of spermatozoa is known as a rather difficult part of semen analysis. Over the past 40 years, morphological criteria became stricter and reference values changed significantly, leading to a 4% lower reference value for normal/typical morphology in 2010. This has consequences for the statistical power of the analysis. Moreover, many laboratories do not use the staining method as advised by the WHO and are getting stricter and stricter in the application of the criteria. Improvement of the assay is therefore necessary. In this study, as a first step, variation in the use of the strict criteria is evaluated. Study design, size, duration Data from the Dutch external quality control (EQC) program were evaluated in this retrospective study over the period 2015 – 2020. The program consists of four rounds per year and includes the assessment of three photos of Papanicolaou stained spermatozoa. These spermatozoa were dichotomously judged (normal/abnormal) on 14 morphological criteria (WHO manual, 2010). Consensus results of three experts served as reference. In total, variation over results of 72 photos (1008 values) was analysed. Participants/materials, setting, methods Participants were staff members from Dutch laboratories (1 member per lab per round) that perform semen analysis. The outcomes of the participants were tested for variation per criterion, both over the entire 6-year period and for trends during this period. To gain insight in the influence of “time”, three photos were provided three times (in 2015/2018/2020) and six photos were provided twice (in 2016/2018 and 2018/2020). Setup was blinded to both participants and experts. Main results and the role of chance In the period 2015 – 2020, 88 – 103 laboratories participated in the EQC program. Of these laboratories, 40 – 60 took part in the photo evaluation. Variation per criterion was expressed in categories green, orange and red, with resp. &amp;gt;90%, 60-90% and &amp;lt;60% agreement between the participants. Overall, variation was in 57% in category green, 37% orange and 6% red. Head ovality, regularity of head contours, regularity of midpiece contours and alignment of the major axis of midpiece and head lead to the most variation. For these criteria, resp. 14, 17, 10 and 17% were in the category red and resp. 50, 47, 71 and 64% in category orange. Lowest variation was found for acrosomal vacuoles, excessive residual cytoplasm, tail thickness and tail length with resp. 76, 77, 94, 85% in category green. Trend analysis lead to similar conclusions: most criteria show a slightly positive trend, but head ovality and regularity of head and midpiece show a stable or declining trend. Three photos were used in three rounds and six photos in two rounds. In 26 (8.8%) cases, shifts towards higher (5) or lower (21) variation were found. Experts changed their opinion in 3 (1%) cases. Limitations, reasons for caution Results are dependent on the morphology of the spermatozoa (magnitude of abnormalities) and of the photo quality. Wider implications of the findings The definitions of the criteria need to be better explained and trained, especially for ovality of the head, regularity of the midpiece and overlap of the longitudinal axes of midpiece and head. Moreover, explanation of the criteria in the light of physiology will probably lead to better evaluations by participants. Trial registration number Not Applicable

The Identification of High Risk APS Patients for Thrombosis with Triple Antiphospholipid Antibody Positivity Is Platform Dependent

The antiphospholipid syndrome (APS) is characterized by thrombosis and/or specific pregnancy morbidity with the persistent presence of antiphospholipid antibodies (aPL). Laboratory criteria include aPL detection by a functional assay referred to as lupus anticoagulant (LAC) or by quantitative solid phase assays measuring anti-β2glycoprotein I (aβ2GPI) and anti-cardiolipin (aCL) IgG/IgM antibodies. Triple positivity (i.e. positivity for LAC, aCL and aβ2GPI antibodies irrespective of the isotype) is associated with a high risk for both a first thrombotic event and recurrence. For these high-risk triple positive patients, a repetitive aPL measurement proved not to be required. Reports from external quality control programs illustrate that commercially available aPL assays produce variable results, probably originating from differences in assay design, epitope exposure and a lack of standardization. On the contrary, the detection of triple positivity was recently suggested to be method and platform independent. In a large multicentre study, we aimed to investigate the variability in triple positivity detection and its clinical association between different aPL platforms.In this study we included 853 patients from seven European medical centers. Patients were classified by the local centers resulting in 259 thrombotic APS patients, 204 diseased controls for thrombosis, 196 patients with autoimmune diseases other than APS and 194 patients without any clinical and laboratory criteria for APS. Four platforms, ACL AcuStar (Instrumentation Laboratory, Bedford, MA, USA), Bioplex 2200 (Bio-Rad Laboratories, Hercules, CA, USA), ImmunoCap EliA (ThermoFisher (Phadia), Waltham, MA, USA) and QUANTA Lite ELISA (Inova Diagnostics, San Diego, CA, USA), were selected based on frequency of use and the willingness of manufacturers to provide their assays. All samples with all assays were tested at one location by a single technician. Lupus anticoagulant positivity was determined by the local center, according to the ISTH guidelines.From the 259 patients diagnosed with thrombotic APS, 202 were positive for LAC. Detection of triple positivity proved to be variable between platforms: 118, 101, 111 and 89 detected with Bioplex, ImmunoCap EliA, AcuStar and QUANTA Lite respectively. To investigate the diagnostic efficacy of triple positivity of the different platforms, we calculated the sensitivity, specificity and odds ratio (OR) for thrombosis. The sensitivity of triple positivity for thrombosis was low and varied from 19% up to 26% between the tested platforms (Figure 1A). However, a high specificity for thrombosis with minor variation from the platforms was obtained ranging from 93% up to 96% (Figure 1A). Looking at the OR for thrombosis, triple positivity was significantly correlated with thrombosis independent of the platform used (Figure 1B). However, the OR for thrombosis ranged from 4.2 up to 5.2 for the different platforms used to detect aβ2GPI and aCL aPL.Taken together, our study confirmed the high-risk profile for thrombosis associated with triple positivity, independent of the platform used to detect aβ2GPI and aCL aPL. Contrary, the number of triple positives is dependent on the platform used, resulting in varying sensitivities of triple positivity for thrombosis over the different platforms. The high risk on recurrent clinical complications in triple positive APS patients often requires them to take indefinite oral anticoagulant treatment. Therefore, to avoid misclassification of high-risk APS patients, standardization/uniformity of solid phase assays for the diagnosis of APS is urgently warranted. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.