Rapid detection of respiratory syncytial virus in nasopharyngeal aspirates by reverse transcription and polymerase chain reaction amplification.
A rapid method for detection of respiratory syncytial virus (RSV) in nasopharyngeal aspirates, involving a combination of reverse transcription and polymerase chain reaction amplification (RT-PCR), has been developed. The RT-PCR assay employs oligonucleotide primers specific for the region of the RSV genome which encodes the F1 subunit of the fusion (F) glycoprotein. Other respiratory viruses do not give a positive reaction. The RT-PCR assay was tested on 202 nasopharyngeal aspirates collected from children with clinical signs of respiratory infection, and the results from RT-PCR were compared with those obtained from virus culture and direct detection by enzyme immunoassay (EIA). RT-PCR results were positive in 118 of 125 samples from which RSV was cultured, as well as in 4 of 7 samples which were culture negative but EIA positive. RT-PCR results were negative in 68 of 70 culture-negative, EIA-negative samples, which included 11 samples from which other respiratory viruses were isolated. The speed, sensitivity (94.6%), and specificity (greater than 97%) of the RT-PCR assay suggest that this technique could be useful for rapid detection of RSV in clinical samples.
Paton
A W
AW
Microbiology Department, Adelaide Children's Hospital, South Australia.
Paton
J C
JC
Lawrence
A J
AJ
Goldwater
P N
PN
Harris
R J
RJ
D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D004279:DNA, Viral / Q000235:genetics; D005069:Evaluation Studies as Topic; D006801:Humans; D007223:Infant; D008969:Molecular Sequence Data; D009305:Nasopharynx / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012194:RNA-Directed DNA Polymerase; D012136:Respiratory Syncytial Viruses / Q000235:genetics / Q000302:isolation & purification*; D012141:Respiratory Tract Infections / Q000175:diagnosis; D010253:Respirovirus Infections / Q000175:diagnosis; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic; D014773:Virology / Q000379:methods
D004279:DNA, Viral; D012194:RNA-Directed DNA Polymerase