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A European multicentre evaluation of detection and typing methods for human enteroviruses and parechoviruses using RNA transcripts

Hayes, A. ; Nguyen, D. ; Andersson, M. ; Antón, A. ; Bailly, J. L. ; Beard, S. ; Benschop, K. S.M. ; Berginc, N. ; Blomqvist, S. and Cunningham, E. , et al. (2019) In Journal of Medical Virology
Abstract

Polymerase chain reaction (PCR) detection has become the gold standard for diagnosis and typing of enterovirus (EV) and human parechovirus (HPeV) infections. Its effectiveness depends critically on using the appropriate sample types and high assay sensitivity as 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 four human EV species (EV-A71, echovirus 30, coxsackie A... (More)

Polymerase chain reaction (PCR) detection has become the gold standard for diagnosis and typing of enterovirus (EV) and human parechovirus (HPeV) infections. Its effectiveness depends critically on using the appropriate sample types and high assay sensitivity as 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 four human EV species (EV-A71, echovirus 30, coxsackie A virus 21, and EV-D68), HPeV3, and specificity controls. Reported results from 48 in-house and 15 commercial assays showed 98% detection frequencies of high copy (1000 RNA copies/5 µL) transcripts. In-house assays showed significantly greater detection frequencies of the low copy (10 copies/5 µL) EV and HPeV transcripts (81% and 86%, respectively) compared with commercial assays (56%, 50%; P = 7 × 10−5). EV-specific PCRs showed low cross-reactivity with human rhinovirus C (3 of 42 tests) and infrequent positivity in the negative control (2 of 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%). Stabilized 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.

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organization
publishing date
type
Contribution to journal
publication status
epub
subject
keywords
enterovirus, enterovirus A71, parechovirus, PCR, RNA transcripts
in
Journal of Medical Virology
publisher
John Wiley and Sons Inc.
external identifiers
  • pmid:31883139
  • scopus:85078274990
ISSN
0146-6615
DOI
10.1002/jmv.25659
language
English
LU publication?
yes
id
b7443ef3-5c9b-48a9-9ebb-e36362c994ec
date added to LUP
2020-02-03 12:51:27
date last changed
2020-10-20 03:40:20
@article{b7443ef3-5c9b-48a9-9ebb-e36362c994ec,
  abstract     = {<p>Polymerase chain reaction (PCR) detection has become the gold standard for diagnosis and typing of enterovirus (EV) and human parechovirus (HPeV) infections. Its effectiveness depends critically on using the appropriate sample types and high assay sensitivity as 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 four human EV species (EV-A71, echovirus 30, coxsackie A virus 21, and EV-D68), HPeV3, and specificity controls. Reported results from 48 in-house and 15 commercial assays showed 98% detection frequencies of high copy (1000 RNA copies/5 µL) transcripts. In-house assays showed significantly greater detection frequencies of the low copy (10 copies/5 µL) EV and HPeV transcripts (81% and 86%, respectively) compared with commercial assays (56%, 50%; P = 7 × 10<sup>−5</sup>). EV-specific PCRs showed low cross-reactivity with human rhinovirus C (3 of 42 tests) and infrequent positivity in the negative control (2 of 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%). Stabilized 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.</p>},
  author       = {Hayes, A. and Nguyen, D. and Andersson, M. and Antón, A. and Bailly, J. L. and Beard, S. and Benschop, K. S.M. and Berginc, N. and Blomqvist, S. and Cunningham, E. and Davis, D. and Dembinski, J. L. and Diedrich, S. and Dudman, S. G. and Dyrdak, R. and Eltringham, G. J.A. and Gonzales-Goggia, S. and Gunson, R. and Howson-Wells, H. C. and Jääskeläinen, A. J. and López-Labrador, F. X. and Maier, M. and Majumdar, M. and Midgley, S. and Mirand, A. and Morley, U. and Nordbø, S. A. and Oikarinen, S. and Osman, H. and Papa, A. and Pellegrinelli, L. and Piralla, A. and Rabella, N. and Richter, J. and Smith, M. and Söderlund Strand, A. and Templeton, K. and Vipond, B. and Vuorinen, T. and Williams, C. and Wollants, E. and Zakikhany, K. and Fischer, T. K. and Harvala, H. and Simmonds, P.},
  issn         = {0146-6615},
  language     = {eng},
  publisher    = {John Wiley and Sons Inc.},
  series       = {Journal of Medical Virology},
  title        = {A European multicentre evaluation of detection and typing methods for human enteroviruses and parechoviruses using RNA transcripts},
  url          = {http://dx.doi.org/10.1002/jmv.25659},
  doi          = {10.1002/jmv.25659},
  year         = {2019},
}