Diagnosis of human metapneumovirus and rhinovirus in patients with respiratory tract infections by an internally controlled multiplex real-time RNA PCR

Autor: Sitha A. Scheltinga, Kate Templeton, Matthias F. C. Beersma, Eric C. J. Claas
Rok vydání: 2004
Předmět:
Time Factors
Rhinovirus
Respiratory System
hMPV
human metapneumoviruses
medicine.disease_cause
Polymerase Chain Reaction
law.invention
law
TCID50%
tissue culture infective dose
Multiplex
Child
Respiratory Tract Infections
Polymerase chain reaction
Paramyxoviridae Infections
Respiratory tract infections
biology
Viral culture
virus diseases
Reference Standards
Infectious Diseases
Internal control
Child
Preschool
Rhinoviruses
RNA
Viral
Adult
Adolescent
Real-time RNA PCR
EAV
equine arteritis virus
Sensitivity and Specificity
Virus
Article
Cell Line
Human metapneumovirus
Virology
Multiplex polymerase chain reaction
medicine
Animals
Humans
Respiratory viruses
Picornaviridae Infections
LRT
lower respiratory tract
Metapneumoviruses
hRV
human rhinoviruses
Multiplex PCR
biology.organism_classification
PIV
parainfluenza virus
URTI
upper respiratory tract infection
Metapneumovirus
RSV
respiratory syncytial virus
Zdroj: Journal of Clinical Virology
ISSN: 1386-6532
Popis: Background: Adequate laboratory diagnosis of human rhinoviruses (hRV) and human metapneumoviruses (hMPV) requires molecular methods as viral culture lacks sensitivity. However, setting up individual PCRs for all respiratory viruses is not practical so preferentially multiplex PCRs are used. Objectives: To develop for routine diagnosis a rapid real-time PCR assay for detection of hRV and hMPV including an internal control in a single tube multiplex reaction using probes carrying different fluorophores to discriminate targets. Study design: The multiplex real-time RNA PCR was optimized to include the internal control virus and a total of 358 respiratory samples from 239 patients taken over a one-year period were analyzed by the multiplex assay. Results: The multiplex assay with co-amplification of the internal control was as sensitive and specific as the individual assays. Application of this assay on clinical samples from 239 patients in a one-year period resulted in an incidence of hRV and hMPV of 41/239 (17.1%) and 6/239 (2.5%), respectively. Inhibition, defined as poor internal control amplification, was detected in 8 (2.2%) samples. Culture was performed on these samples and only four hRV were detected. Conclusions: This real-time PCR method enables sensitive diagnosis of these two respiratory pathogens with the potential to expand the assay as part of a full molecular respiratory viral screen.
Databáze: OpenAIRE
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