Reverse transcription polymerase chain reaction and electrospray ionization mass spectrometry for identifying acute viral upper respiratory tract infections.
Diagnosis of respiratory viruses traditionally relies on culture or antigen detection. We aimed to demonstrate capacity of the reverse transcription polymerase chain reaction/electrospray ionization mass spectrometry (RT-PCR/ESI-MS) platform to identify clinical relevant respiratory viruses in nasopharyngeal aspirate (NPA) samples and compare the diagnostic performance characteristics relative to conventional culture- and antigen-based methods. An RT-PCR/ESI-MS respiratory virus surveillance kit designed to detect respiratory syncytial virus, influenza A and B, parainfluenza types 1-4, Adenoviridae types A-F, Coronaviridae, human bocavirus, and human metapneumovirus was evaluated using both mock-ups and frozen archived NPA (N = 280), 95 of which were positive by clinical virology methods. RT-PCR/ESI-MS detected 74/95 (77.9%) known positive samples and identified an additional 13/185 (7%) from culture-negative samples. Viruses that are nondetectable with conventional methods were also identified. Viral load was semiquantifiable and ranged from 2400 to >320 000 copies/mL. Time to results was 8 h. RT-PCR/ESI-MS showed promise in rapid detection of respiratory viruses and merits further evaluation for use in clinical settings.
Department of Emergency Medicine, Johns Hopkins University, Baltimore, MD 21204, USA. kfchen@adm.cgmh.org.tw
Kuan-Fu
KF
Chen
Lawrence
L
Blyn
Richard E
RE
Rothman
Padmini
P
Ramachandran
Alexandra
A
Valsamakis
David
D
Ecker
Rangarajan
R
Sampath
Charlotte A
CA
Gaydos
D000208:Acute Disease; D000256:Adenoviridae / Q000235:genetics; D000465:Algorithms; D006801:Humans; D009980:Influenza A virus / Q000235:genetics; D009981:Influenza B virus / Q000235:genetics; D010223:Parainfluenza Virus 2, Human / Q000235:genetics; D010224:Parainfluenza Virus 3, Human / Q000235:genetics; D011933:Reagent Kits, Diagnostic / Q000592:standards; D015203:Reproducibility of Results; D012136:Respiratory Syncytial Viruses / Q000235:genetics; D012141:Respiratory Tract Infections / Q000175:diagnosis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization
D011933:Reagent Kits, Diagnostic
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