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A European multi-centre evaluation of detection and typing methods for human enteroviruses and parechoviruses using RNA transcripts

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  • A Hayes
  • D Nguyen
  • M Andersson
  • A Antón
  • J-L Bailly
  • S Beard
  • K S M Benschop
  • N Berginc
  • S Blomqvist
  • E Cunningham
  • D Davis
  • J L Dembinski
  • S Diedrich
  • S G Dudman
  • R Dyrdak
  • G J A Eltringham
  • S Gonzales-Goggia
  • R Gunson
  • H C Howson-Wells
  • A J Jääskeläinen
  • F X López-Labrador
  • M Maier
  • M Majumdar
  • S Midgley
  • A Mirand
  • U Morley
  • S A Nordbø
  • S Oikarinen
  • H Osman
  • A Papa
  • L Pellegrinelli
  • A Piralla
  • N Rabella
  • J Richter
  • M Smith
  • A Söderlund Strand
  • K Templeton
  • B Vipond
  • T Vuorinen
  • C Williams
  • E Wollants
  • K Zakikhany
  • T K Fischer
  • H Harvala
  • P Simmonds
Vis graf over relationer

PCR detection has become the gold standard for diagnosis and typing of enterovirus (EVs) and human parechovirus (HPeV) infections. Its effectiveness depends critically on using appropriate sample types and high assay sensitivity since viral loads in cerebrospinal fluid samples from meningitis and sepsis clinical presentation can be extremely low. This study evaluated the sensitivity and specificity of currently used commercial and in-house diagnostic and typing assays. Accurately quantified RNA transcript controls were distributed to 27 diagnostic and 12 reference laboratories in 17 European countries for blinded testing. Transcripts represented the 4 human EV species (EV-A71, echovirus 30, coxsackie A virus 21, EV-D68), HPeV-3 and specificity controls. Reported results from 48 in-house and 15 commercial assays showed 98% detection frequencies of high copy (1000 RNA copies/5µl) transcripts. In-house assays showed significantly greater detection frequencies of the low copy (10 copies/5µl) EV and HPeV transcripts (81%, 86% respectively) compared to commercial assays (56%, 50%; p = 7x10-5 ). EV-specific PCRs showed low cross-reactivity with human rhinovirus C (3/42 tests) and infrequent positivity in the negative control (2/63 tests). Most or all high copy EV and HPeV controls were successfully typed (88%, 100%) by reference laboratories, but showed reduced effectiveness for low copy controls (41%, 67%). Stabilised RNA transcripts provide an effective, logistically simple and inexpensive reagent for evaluation of diagnostic assay performance. The study provides reassurance of the performance of the many in-house assay formats used across Europe. However, it identified often substantially reduced sensitivities of commercial assays often used as point-of-care tests. This article is protected by copyright. All rights reserved.

OriginalsprogEngelsk
TidsskriftJournal of Medical Virology
Vol/bind92
Udgave nummer8
Sider (fra-til)1065-1074
ISSN0146-6615
DOI
StatusUdgivet - aug. 2020

Bibliografisk note

This article is protected by copyright. All rights reserved.

ID: 58860494