Duplex real-time RT-PCR assay for detection and subgroup-specific identification of human respiratory syncytial virus.
DNA Primers
/ genetics
DNA Probes
/ genetics
Humans
Limit of Detection
Nasopharynx
/ virology
RNA, Viral
/ isolation & purification
Real-Time Polymerase Chain Reaction
Reproducibility of Results
Respiratory Syncytial Virus Infections
/ diagnosis
Respiratory Syncytial Virus, Human
/ classification
Sensitivity and Specificity
HRSV
HRSV subgroups
Human respiratory syncytial virus
Real-time RT-PCR
Respiratory viruses
Journal
Journal of virological methods
ISSN: 1879-0984
Titre abrégé: J Virol Methods
Pays: Netherlands
ID NLM: 8005839
Informations de publication
Date de publication:
09 2019
09 2019
Historique:
received:
01
03
2019
revised:
06
06
2019
accepted:
07
06
2019
pubmed:
11
6
2019
medline:
4
6
2020
entrez:
11
6
2019
Statut:
ppublish
Résumé
Human respiratory syncytial virus (HRSV) is a leading cause of acute respiratory illness in young children worldwide. Reliable detection and identification of HRSV subgroup A and B infections are essential for accurate disease burden estimates in anticipation of licensure of novel HRSV vaccines and immunotherapies. To ensure continued reliability, molecular assays must remain current with evolving virus strains. We have developed a HRSV subgroup-specific real-time RT-PCR (rRT-PCR) assay for detection and subgroup identification using primers and subgroup-specific probes targeting a conserved region of the nucleoprotein gene combined in a single duplex reaction using all genome sequence data currently available in GenBank. The assay was validated for analytical sensitivity, specificity, reproducibility, and clinical performance with a geographically diverse collection of viral isolates and respiratory specimens in direct comparison with an established pan-HRSV rRT-PCR reference test. The assay was sensitive, reproducibly detecting as few as 5-10 copies/reaction of target RNA. The assay was specific, showing no amplification with a panel of 16 other common respiratory pathogens or predicted by in silico primer/probe analysis. The duplex rRT-PCR assay based on the most current available genome sequence data permits rapid, sensitive and specific detection and subgroup identification of HRSV.
Identifiants
pubmed: 31181218
pii: S0166-0934(19)30072-2
doi: 10.1016/j.jviromet.2019.113676
pmc: PMC7172218
pii:
doi:
Substances chimiques
DNA Primers
0
DNA Probes
0
RNA, Viral
0
Types de publication
Journal Article
Validation Study
Langues
eng
Sous-ensembles de citation
IM
Pagination
113676Informations de copyright
Copyright © 2019 Elsevier B.V. All rights reserved.
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