Simultaneous detection of respiratory viruses in children with acute respiratory infection using two different multiplex reverse transcription-PCR assays.
A 4-tube multiplex RT-PCR (mRT-PCR), which showed higher sensitivity over conventional methods, was previously developed for the diagnosis of 14 viral pathogens of the respiratory tract. Herein the mRT-PCR was compared to the commercial Luminex mPCR-microsphere flow cytometry assay (Resplex II) which allows the detection of 12 different viruses. Eleven different viruses were identified in 91 nasopharyngeal swabs of children with acute respiratory infection, influenza A (IAV) and B, respiratory syncytial virus (RSV), human rhinovirus (hRhV), human echovirus, parainfluenza viruses (PIV) 1, 2, 3 and 4, human metapneumovirus (hMPV), and human coronavirus NL63. The results of the two techniques showed 53 and 40 positive patients by the Resplex II assay and mRT-PCR, respectively, with a concordance in 35 positive and 33 negative patients (74.7%). Individual RT-PCR tests were performed to control viruses not simultaneously detected by the two multiplex assays. The major virus misdiagnosed by mRT-PCR was IAV whereas the major viruses misdiagnosed by Resplex II were PIV1, 3 and 4. The mRT-PCR remains a simple, rapid, and specific assay for the specific detection of respiratory viruses, and can be easily implemented with standards in clinical laboratories at a low cost.
Wang
Wei
W
Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China.
Ren
Peijun
P
Sheng
Jun
J
Mardy
Sek
S
Yan
Huajie
H
Zhang
Jing
J
Hou
Lili
L
Vabret
Astrid
A
Buchy
Philippe
P
Freymuth
Francois
F
Deubel
Vincent
V
D002648:Child; D002675:Child, Preschool; D005434:Flow Cytometry; D006801:Humans; D007223:Infant; D009305:Nasopharynx / Q000821:virology; D012327:RNA Virus Infections / Q000175:diagnosis / Q000821:virology; D012328:RNA Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification; D012141:Respiratory Tract Infections / Q000175:diagnosis / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014780:Viruses
D012367:RNA, Viral