scholarly journals Inadequate design of mutation detection panels prevents interpretation of variants of concern: results of an external quality assessment for SARS-CoV-2 variant detection

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Christoph Buchta ◽  
Jeremy V. Camp ◽  
Jovana Jovanovic ◽  
Ulla Radler ◽  
Bernhard Benka ◽  
...  
Keyword(s):  
Quality Assessment ◽  
Mutation Detection ◽  
External Quality Assessment ◽  
Melting Curve ◽  
Laboratory Diagnostics ◽  
External Quality ◽  
Specific Pcr ◽  
High Throughput Method ◽  
Individual Variant ◽  
Selection Of

Abstract Objectives Mutation-specific PCR assays have quickly found their way into laboratory diagnostics due to their capacity to be a fast, easy to implement and high-throughput method for the detection of known SARS-CoV-2 variants of concern (VoCs). However, little is known about the performance of such assays in routine laboratory analysis. Methods The results reported in a recent round of an external quality assessment (EQA) scheme for SARS-CoV-2 mutation-specific PCR were retrospectively analyzed. For the determination of individual variant-specific sequences as well as for the interpretation results for certain virus variants, correct, incorrect, and unreported results were evaluated, and their possible causes were investigated. Results A total of 34 laboratories participated in this study. For five samples containing the VoC Alpha + E484K, Beta, Gamma, Delta, or B.1.1.318 (as a variant of interest), 848 results for SARS-2-CoV mutation detection were reported, 824 (97.2%, range per sample 88–100%) of which were correct. Melting curve assays gave 99% correct results, real-time RT-qPCR 94%, microarray-based assays 100%, and MALDI-TOF MS 96%. A total of 122/167 (73%) reported results for SARS-CoV-2 variant determination were correct. Of the 45 inconclusive or incorrect results, 33 (73%) were due to inadequate selection of targets that did not allow identification of contemporary VoC, 11 (24%) were due to incorrect results, and one (3%) was due to correct results of mutation-specific PCR. Conclusions Careful and up-to-date selection of the targets used in mutation-specific PCR is essential for successful detection of current SARS-CoV-2 variants.

Anti-ganglioside antibodies: experience from the Italian Association of Neuroimmunology external quality assessment scheme

2018 ◽  
Vol 56 (11) ◽  
pp. 1921-1925 ◽  
Author(s):  
Diego Franciotta ◽  
Matteo Gastaldi ◽  
Tiziana Biagioli ◽  
Luana Benedetti ◽  
Claudia Giannotta ◽  
...  
Keyword(s):  
Quality Assessment ◽  
External Quality Assessment ◽  
Real Life ◽  
Laboratory Diagnostics ◽  
Serum Samples ◽  
External Quality ◽  
External Quality Assessment Scheme ◽  
Italian Association ◽  
Commercial Line ◽  
Ganglioside Antibodies

Abstract Background Anti-ganglioside antibodies are currently used in the differential diagnosis of suspected immune-mediated neuropathies. In-house and increasingly used commercial assays seem to perform suboptimally, and comparative information on their analytical performance are essentially lacking. Born within the frame of guidelines and standardization activities by the Italian Association of Neuroimmunology, this external quality assessment scheme (EQAS) is a real-life snapshot of the laboratory diagnostics in this field. Methods The EQAS consisted of five surplus, anonymized serum samples from patients with clinically-defined neuropathies and two serum samples from healthy blood donors. Eight laboratories used commercial line-/dot-blots, seven in-house/commercial ELISAs (in addition, 13 laboratories tested a recently released ELISA by Bühlmann). Only high anti-ganglioside antibody reactivities were considered, in accordance with consolidated recommendations. Results Large variations in anti-ganglioside antibody profiles were observed, even, although to a lesser extent, within homogeneous classes of assays. Concordance between the profiles and clinical phenotypes was also partial. Conclusions Although conducted on a relatively small, but representative number of Italian laboratories, this EQAS shows a critical between-laboratory disagreement in the test results of anti-ganglioside antibodies. Also considering the trend for using certified assays in generalist laboratories, strong efforts toward standardization and the identification of the best method(s) for their determinations are compellingly needed.

scholarly journals External quality assessment for molecular diagnostic laboratories in Belgium: Can we improve it?

2019 ◽  
Vol 25 (1) ◽  
pp. 39-49
Author(s):  
Kelly Dufraing ◽  
Els Lierman ◽  
Anne Vankeerberghen ◽  
Sabine Franke ◽  
Els Dequeker
Keyword(s):  
Quality Assessment ◽  
Performance Monitoring ◽  
External Quality Assessment ◽  
Molecular Diagnostic ◽  
Real Patient ◽  
External Quality ◽  
Molecular Tests ◽  
Clinical Biology ◽  
Eqa Schemes ◽  
Selection Of

AbstractExternal quality assessment (EQA) is an essential part of performance monitoring for molecular laboratories. At the moment, a national law regulates participation in EQA schemes for clinical biology and pathology in Belgium. This study aimed (1) to get insights on how laboratories organize their EQA participation, (2) to poll satisfaction with the current situation (selection of EQA programs in advance by a governmental body), (3) to provide guidance for choosing the most relevant EQA provider and (4) to propose a new model for national performance monitoring. A survey was sent to Belgian laboratories performing molecular tests in the field of microbiology, hematology and pathology with (1) general questions on how they select an EQA provider and (2) their satisfaction of each provider. In total, 25 molecular laboratories [microbiology (N = 13), hematology (N = 8) and pathology (N = 4)] from 14 different hospitals completed the survey regarding their EQA organization. All three laboratory groups indicated to prefer EQA schemes using real patient materials as well as those with varying targets and concentrations. For molecular microbiology and hematology, schemes with a syndromic approach are sought. Since annual participation in EQA becomes burdensome in most laboratories, this paper also offers a risk-based strategy for determining the participation frequency. Based on the needs of Belgian laboratories, three proposals were made: (1) for the proper selection of an EQA scheme, (2) for determining the minimal participation frequency and (3) for the national organization of EQA schemes.

Results of the first external quality assessment scheme (EQA) for isolation and analysis of circulating tumour DNA (ctDNA)

2018 ◽  
Vol 56 (2) ◽  
pp. 220-228 ◽  
Author(s):  
Verena Haselmann ◽  
Parviz Ahmad-Nejad ◽  
Wolf J. Geilenkeuser ◽  
Angelika Duda ◽  
Merle Gabor ◽  
...  
Keyword(s):  
Quality Assessment ◽  
Error Rate ◽  
External Quality Assessment ◽  
Digital Pcr ◽  
Laboratory Diagnostics ◽  
External Quality ◽  
External Quality Assessment Scheme ◽  
Genotyping Platform ◽  
Edta Plasma ◽  
Circulating Tumour Dna

Abstract Background: Circulating tumour DNA (ctDNA) is considered to have a high potential for future management of malignancies. This pilot external quality assessment (EQA) scheme aimed to address issues of analytical quality in this new area of laboratory diagnostics. Methods: The EQA scheme consisted of three 2-mL EDTA-plasma samples spiked with fragmented genomic DNA with a mutant allele frequency ranging from 0% to 10% dedicated to the analysis of nine known sequence variations in KRAS codon 12/13 and of BRAF V600E. Laboratories reported: (1) time elapsed for processing, (2) storage temperatures, (3) methods for extraction and quantification, (4) genotyping methodologies and (5) results. Results: Specimens were sent to 42 laboratories from 10 European countries; 72.3% reported to isolate cell-free DNA (cfDNA) manually, 62.5% used the entire plasma volume for cfDNA isolation and 38.5% used >10% of cfDNA extracted for downstream genotyping. Of the methods used for quantification, PicoGreen demonstrated the lowest coefficient of variation (33.7%). For genotyping, 11 different methods were reported with the highest error rate observed for Sanger sequencing and the lowest for highly sensitive approaches like digital PCR. In total, 197 genotypes were determined with an overall error rate of 6.09%. Conclusions: This pilot EQA scheme illustrates the current variability in multiple phases of cfDNA processing and analysis of ctDNA resulting in an overall error rate of 6.09%. The areas with the greatest variance and clinical impact included specimen volume, cfDNA quantification method, and preference of genotyping platform. Regarding quality assurance, there is an urgent need for harmonisation of procedures and workflows.

External quality assessment programs of laboratory test results conducted by the Central Center for Quality Research in Laboratory Diagnostics in 2019

2020 ◽  
Vol 56 (2) ◽  
pp. 1-20
Author(s):  
Barbara Przybył-Hac ◽  
Jadwiga Ciechowicz ◽  
Dorota Waszczuk-Łysiuk
Keyword(s):  
Quality Assessment ◽  
External Quality Assessment ◽  
Clinical Chemistry ◽  
Laboratory Diagnostics ◽  
Test Results ◽  
Acid Base Balance ◽  
External Quality ◽  
Acceptable Error ◽  
Quality Research ◽  
Laboratory Test Results

In 2019, the Central Center for Quality Research in Laboratory Diagnostics conducted 24 editions as part of eight external quality assessment programs. Four programs (in the field of clinical chemistry, hematology, parameters of acid-base balance and ISE electrolytes, and coagulatory) were conducted with the frequency of four editions a year, which gradually adjusts the implementation of the programs to the requirements of the Regulation of the Minister of Health of December 15, 2017 (Journal of Laws No. U. 2017, item 2394). The other four programs, for financial reasons, were conducted with the frequency of two editions a year. In 2019, the Central Center also maintained a procedure for transporting control material samples to laboratories by courier. In total, 558.947 control test results from 1542 polish medical diagnostic laboratories were assessed in 2019. In seven programs carried out in 2019, the percentage of correct results according to the adopted group criteria was slightly higher compared to 2018. Only in the hematology program, the percentage of correct results according to group means was lower due to the applied limits of the acceptable error limits. It is worth emphasizing thatthe annual regularity assessments obtained in 2019 are comparable to those obtained in 2018.

scholarly journals Variable sensitivity in molecular detection of SARS-CoV-2 in European Expert Laboratories: External Quality Assessment, June – July 2020

2020 ◽  
Author(s):  
Carlo Fischer ◽  
Ramona Mögling ◽  
Angeliki Melidou ◽  
Arne Kühne ◽  
Edmilson F. Oliveira-Filho ◽  
...  
Keyword(s):  
Quality Assessment ◽  
Molecular Detection ◽  
External Quality Assessment ◽  
External Quality ◽  
Human Influenza Virus ◽  
Robust Detection ◽  
Variable Sensitivity ◽  
Human Coronaviruses ◽  
June July ◽  
Selection Of

Objective: During the ongoing coronavirus disease 2019 (COVID-19) outbreak, robust detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a key element for clinical management and to interrupt transmission chains. We organized an external quality assessment (EQA) of molecular detection of SARS-CoV-2 for European expert laboratories. Study design: An EQA panel composed of 12 samples, containing either SARS-CoV-2 at different concentrations to evaluate sensitivity or other respiratory viruses to evaluate specificity of SARS-CoV-2 testing, was distributed to 68 laboratories in 35 countries. Specificity samples included seasonal human coronaviruses hCoV-229E, hCoV-NL63 and hCoV-OC43, as well as MERS-CoV, SARS-CoV and human influenza virus A and B. Conclusion: Sensitivity was variable among laboratories, particularly for low concentration SARS-CoV-2 samples. Results indicated that performance was mostly independent of the selection of specific extraction or PCR methods.

External quality assessment programs of laboratory test results conducted by the Central Center for Quality Research in Laboratory Diagnostics in 2018

2019 ◽  
Vol 55 (3) ◽  
pp. 209-228 ◽  
Author(s):  
Barbara Przybył-Hac ◽  
Jadwiga Ciechowicz ◽  
Dorota Waszczuk-Łysiuk
Keyword(s):  
Quality Assessment ◽  
Glycated Hemoglobin ◽  
External Quality Assessment ◽  
Laboratory Diagnostics ◽  
Acid Base Balance ◽  
External Quality ◽  
Quality Research ◽  
Laboratory Test Results ◽  
Base Balance ◽  
First Time

In 2018, the Center for Quality Research in Laboratory Diagnostics conducted 22 editions under eight external quality assessment programs. In two programs (in the field of hematology and parameters of acid-base balance and electrolytes of ISE) the frequency of editing from two to four has been increased, gradually adapting the programs to the requirements of the Regulation of the Minister of Health of December 15, 2017 (Journal of Laws). As part of the extension of the program offer, the Central Center introduced 12 new analytes in the coagulology program and implemented a new external quality assessment program for HbA1c glycated hemoglobin controls. The Central Center also implemented in 2018 a new procedure for transporting samples of control materials to laboratories. For the first time, the parcels were delivered by courier. In all programs implemented in 2018, higher percentages of correct results were obtained according to the adopted criteria and annual regularity assessments comparable to 2017. Very satisfactory results were also obtained for glycated hemoglobin, HbA1c – a pilot program implemented in 2018.

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