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

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. >90%, 60-90% and <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

Питання зовнішнього контролю якості лабораторних досліджень на сифіліс у лікувальних закладах України

The objective of the work is to identify systemic and random errors in the formulation of controlled methods and to achieve comparative results obtained by laboratories participating in external quality control of laboratory tests for syphilis. Materials and methods. Laboratories of three levels took part in the activities of external quality control of serological diagnostics of syphilis: clinical diagnostic laboratories (CDL) of medical institutions of scientific research institutes of the NAMS of Ukraine, serological departments as part of the CDL of the dermatovenerological profile of the RDVD, CDVD. Results. The quality of serological tests for syphilis in 21 CDL (serological departments) was assessed using the following methods: RMP, RPR, Rapid-test, TPHA, FTA-abs, ELISA in three control serum samples according to formal and essential characteristics. The most significant differences in the results were revealed in the study of control serum samples with a low content of antibodies and those without antibodies to the causative agent of syphilis in the non-treponemal RMP test and the treponemal FTA-abs test. Conclusions. The conducted studies have shown the high efficiency of the practical application of the developed system of external quality control of laboratory methods for diagnosing syphilis.

Development of the optimal procedure for increasing HbA1c concentration in control materials for external quality assessment

Background: The research aimed to increase certain HbA1c concentrations at medical decision levels for external quality control samples from healthy donor blood. Methods: The in vitro study was performed from October 2019 to January 2021 at Quality Control Center for Medical Laboratory at University of Medicine and Pharmacy at Ho Chi Minh City. The study observed on the conditions including the optimal buffer solutions (BAGPM, BPS, Ringer, Saline), temperature (2ºC - 8ºC, 22ºC - 24ºC, 37ºC), and glucose concentration (100 mM, 305 mM, 500 mM) affecting the HbA1c concentration in vitro to make the external quality control samples fell in normal, prediabetes, and diabetes range. At every condition, the HbA1c concentration was measured by Tina Quant method to look for the optimal procedure to increase HbA1c concentration required of the external quality control protocol. Results: The highest HbA1c concentration (11.57±0.2%) was found in BAGPM solution with 100 mM glucose after 15 days with the baseline HbA1c 5.43±0.13%; the HbA1c level increase dramatically at 37ºC in BAGPM 500 mM glucose solution in fifteen days (40.03±1.05%). Conclusions: The appropriate conditions were identified to prepare HbA1c standards for prediabetic and diabetic levels. The standards for HbA1c concentrations were recommended to prepare by incubating RBCs from non-diabetic donor blood in BAGPM solution containing glucose at 37ºC for 24 hours. Glucose concentrations should be 100 mM and 500 mM, respectively, for prediabetic level (HbA1c ~ 6.0 ± 0.12%) and diabetic level (HbA1c ~ 9.6 ± 0.17%).

Aspergillus Polymerase Chain Reaction—An Update on Technical Recommendations, Clinical Applications, and Justification for Inclusion in the Second Revision of the EORTC/MSGERC Definitions of Invasive Fungal Disease

Aspergillus polymerase chain reaction testing of blood and respiratory samples has recently been included in the second revision of the EORTC/MSGERC definitions for classifying invasive fungal disease. This is a result of considerable efforts to standardize methodology, the availability of commercial assays and external quality control programs, and additional clinical validation. This supporting article provides both clinical and technical justifications for its inclusion while also summarizing recent advances and likely future developments in the molecular diagnosis of invasive aspergillosis.

Internal audit key quality components

The services provided by internal auditors are based on a high level of confidence in the quality of their work. Continuous internal and periodic external quality control of internal audit activities are of key importance for maintaining confidence in the work of internal auditors.

Performance of the Boditech iCHROMA Covid-19 IgG antibody assay with the external quality control for UK NIBSC (National Institute of Biological Standards and Control)

Background: There is a need for laboratories to participate in EQA schemes to achieve a harmonization of test results and to improve serological testing and research on COVID-19 and SARS-CoV-2. The Boditech iCHROMA point of care immunoassay analyser has a Covid-19 antibody assay for the qualitative determination of IgG antibodies in human whole blood/serum/plasma. The National Institute for Biological Standards and Control (NIBSC) in the United Kingdom has a portfolio of reference and research reagents, including reagents for EQA that are available to laboratories to help them set up and develop accurate diagnostic tests for SARS-CoV-2 assays. Objectives: To determine the performance of the Boditech iCHROMA Covid-19 IgG antibody assay using the external quality control material provided by the UK NIBSC (National Institute for Biological Standards and Control). Results: The Boditech iCHROMA Covid-19 IgG assay using the cut off index >1.1, reported the results of the 33 positive control samples as positive with index range from 19.50 – 29.00, with a mean of 23.75 and standard deviation of SD of 2.53. It also reported the results of the 33 negative control samples as negative with an index range of 0.00 for all samples. Conclusion: The performance of the Boditech iCHROMA Covid-19 IgG antibody assay using external quality control material from the UK NIBSC was very good, confirming the precision of the Boditech iCHROMA Covid 19 IgG antibody method.

Tuning of the proficiency testing methodology for soil fertility analysis. Preliminary results

This work includes the methodology for carrying out a proficiency testing program between Spanish laboratories that offer soil fertility analysis. Aliquots of each of the two large volume samples were prepared, taken from two surface horizons of soils with marked differences between them. Once the homogeneity of aliquots was tested, one sample of each soil was sent to each of the twenty-one participating laboratories, together with the instructions and a format for the submission of results. Throughout the process, special care was taken to ensure the confidentiality of the results of each individual participant. Once the responses were received, the results were analyzed, consensus values were assigned and each result was evaluated using the “z-score” indicator. The exercise was very well received by the participating laboratories, and has been very effective in characterizing the quality of the results issued. It has been shown that there are parameters with very homogeneous results (pH) compared to others with differences incompatible with the reliable agronomic use of analyses (granulometry, assimilable phosphorus, etc.). The urgent need to maintain this type of intercomparison exercise periodically, as an external quality control that increases the reliability of laboratories, is evident.