Use of a diagnostic Puumala virus real-time RT-PCR in an orthohantavirus endemic region in the Netherlands

Laboratory diagnosis of orthohantavirus infection is primarily based on serology. However, for a confirmed serological diagnosis, evaluation of a follow-up serum sample is essential, which is time consuming and causes delay. Real-time reverse transcription polymerase chain reaction (RT-PCR) tests, i...

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Bibliographic Details
Published in:	Microbiology spectrum Vol. 12; no. 7; p. e0381323
Main Authors:	Geeraedts, Felix, Wevers, Mariska, Bosma, Froukje, Boer, Maria de, Brinkman, J N, Delsing, Corine, GeurtsvanKessel, Corine, Rockx, Barry, van der Zanden, Adri, Laverman, Gozewijn D
Format:	Journal Article
Language:	English
Published:	United States American Society for Microbiology 02-07-2024
Subjects:	Antibodies, Viral - blood Clinical Microbiology Endemic Diseases Female hantavirus Hantavirus Infections - diagnosis Hantavirus Infections - epidemiology Hantavirus Infections - virology Hemorrhagic Fever with Renal Syndrome - diagnosis Hemorrhagic Fever with Renal Syndrome - epidemiology Hemorrhagic Fever with Renal Syndrome - virology Humans Immunoglobulin M - blood Male molecular methods Netherlands - epidemiology nucleic acid amplification test Orthohantavirus - classification Orthohantavirus - genetics Orthohantavirus - isolation & purification Puumala virus Puumala virus - genetics Puumala virus - isolation & purification Real-Time Polymerase Chain Reaction - methods Research Article Reverse Transcriptase Polymerase Chain Reaction - methods RNA, Viral - genetics Serologic Tests - methods serology zoonotic infections Netherlands molecular methods nephrology diagnostics Puumala virus zoonotic infections hantavirus serology nucleic acid amplification test
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Summary:	Laboratory diagnosis of orthohantavirus infection is primarily based on serology. However, for a confirmed serological diagnosis, evaluation of a follow-up serum sample is essential, which is time consuming and causes delay. Real-time reverse transcription polymerase chain reaction (RT-PCR) tests, if positive, provide an immediate and definitive diagnosis, and accurately identify the causative agent, where the discriminative nature of serology is suboptimal. We re-evaluated sera from orthohantavirus-suspected clinical cases in the Dutch regions of and from July 2014 to April 2016 for the presence of Puumala orthohantavirus (PUUV), Tula orthohantavirus (TULV), and Seoul orthohantavirus (SEOV) RNA. PUUV RNA was detected in 11% of the total number ( = 85) of sera tested, in 50% of sera positive for anti-PUUV/TULV IgM ( = 16), and in 1.4% of sera negative or indeterminate for anti-PUUV/TULV IgM ( = 69). No evidence was found for the presence of TULV or SEOV viral RNA. Based on these findings, we propose two algorithms to implement real-time RT-PCR testing in routine orthohantavirus diagnostics, which optimally provide clinicians with early confirmed diagnoses and could prevent possible further invasive testing and treatment. The addition of a real-time reverse transcription polymerase chain reaction test to routine orthohantavirus diagnostics may better aid clinical decision making than the use of standard serology tests alone. Awareness by clinicians and clinical microbiologists of this advantage may ultimately lead to a reduction in over-hospitalization and unnecessary invasive diagnostic procedures.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Present address: Viroscience, Erasmus University Medical Center, Rotterdam, Zuid-Holland, the Netherlands The authors declare no conflict of interest.
ISSN:	2165-0497 2165-0497
DOI:	10.1128/spectrum.03813-23