20.500.12556/RUL-153975 Sporadic hepatitis A virus PCR false-positive results observed during reflex testing of serum samples previously tested for Anti-HAV antibodies and caused by contamination with HAV RNA present in the reagents of the commercial Anti-HAV immunoassay Hepatitis A diagnosis relies on serology and occasionally on hepatitis A virus (HAV) RNA detection. For timely diagnosis and the avoidance of drawing additional blood, molecular testing is often performed as reflex testing by using blood specimens that were initially sent for anti-HAV serology. Reflex molecular testing is preferably performed from different sample aliquots, but, for limited sample quantities, it uses samples that have been preprocessed in an immunoassay analyzer. In 2012, we first observed sporadic HAV RNA-positive cases that were inconsistent with patients’ serological profiles and/ or medical histories, suggesting that occasional laboratory contamination was causing false-positive PCR results. Multiple external quality assurance (EQA) and laboratory surface contamination checks were performed, questionable specimens were tested with various HAV RNA tests, and follow-up serum/stool samples were collected. All contamination-check samples and samples from healthy individuals tested HAV RNA-negative, and the laboratory successfully passed all EQAs. The HAV RNA-positive results were reproducible with various HAV RNA assays. No patients seroconverted, and their follow-up samples were consistently HAV RNA-negative. Finally, a detailed review of testing protocols revealed a correlation between HAV RNA false positivity and preceding anti-HAV testing with the Cobas-e411 automated immunoassay analyzer. HAV RNA was detected in the Cobas-e411 anti-HAV reagents, with the HAV sequences matching those from the false-positive samples. Preceding anti-HAV testing using two other immunoassay analyzers did not result in subsequent HAV RNA false positivity during reflex testing. The Cobas-e411 pipetting procedure with a single pipette tip collecting samples and anti-HAV reagents contaminated the original sample with the HAV RNA that was present in the immunoassay’s reagents, thereby resulting in HAV RNA false positivity during the reflex testing. hepatitis A virus luxS intercellular signaling HAV RNA contamination diagnostics serology reflex testing patogeni mikroorganizmi Campylobacter jejuni luxS true false true Angleški jezik Slovenski jezik Članek v reviji 2024-01-17 12:30:49 2024-01-17 12:30:56 2024-01-18 03:42:11 0000-00-00 00:00:00 2023 0 0 6 str. iss. 3 Vol. 11 2023 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 579 2165-0497 10.1128/spectrum.00122-23 151706883 lunar-et-al-2023-sporadic-hepatitis-a-virus-pcr-false-positive-results-observed-during-reflex-testing-of-serum-samples_(1).pdf lunar-et-al-2023-sporadic-hepatitis-a-virus-pcr-false-positive-results-observed-during-reflex-testing-of-serum-samples_(1).pdf 1 C36A5D808DFD280E1D215BA7F351538C 7999e5d5ab11efba1e2785f6169812c88553a91a6c16141b5c4e01ec562c4940 61f90c45-b52c-11ee-b3d9-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=180069 https://journals.asm.org/doi/10.1128/spectrum.00122-23 0 2ae727a3-b52c-11ee-b3d9-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=180068 Medicinska fakulteta 0 0 0