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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
ID
Lunar, Maja M.
(
Author
),
ID
Markočič, Petra
(
Author
),
ID
Fujs Komloš, Kristina
(
Author
),
ID
Štamol, Tina
(
Author
),
ID
Poljak, Mario
(
Author
)
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URL - Source URL, Visit
https://journals.asm.org/doi/10.1128/spectrum.00122-23
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Abstract
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.
Language:
English
Keywords:
hepatitis A virus
,
luxS
,
intercellular signaling
,
HAV RNA
,
contamination
,
diagnostics
,
serology
,
reflex testing
Work type:
Article
Typology:
1.01 - Original Scientific Article
Organization:
MF - Faculty of Medicine
Publication status:
Published
Publication version:
Version of Record
Year:
2023
Number of pages:
6 str.
Numbering:
Vol. 11, iss. 3
PID:
20.500.12556/RUL-153975
UDC:
579
ISSN on article:
2165-0497
DOI:
10.1128/spectrum.00122-23
COBISS.SI-ID:
151706883
Publication date in RUL:
17.01.2024
Views:
549
Downloads:
29
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Record is a part of a journal
Title:
Microbiology spectrum
Shortened title:
Microbiol. spectr.
Publisher:
ASM Press
ISSN:
2165-0497
COBISS.SI-ID:
4526968
Licences
License:
CC BY 4.0, Creative Commons Attribution 4.0 International
Link:
http://creativecommons.org/licenses/by/4.0/
Description:
This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.
Secondary language
Language:
Slovenian
Keywords:
patogeni mikroorganizmi
,
Campylobacter jejuni
,
luxS
Projects
Funder:
Other - Other funder or multiple funders
Funding programme:
University of Ljubljana, Faculty of Medicine, Institute of Microbiology and Immunology
Funder:
ARRS - Slovenian Research Agency
Project number:
P3-0083
Name:
Odnosi parazitskega obstajanja
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