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Journal of Clinical Microbiology, November 1998, p. 3149-3154, Vol. 36, No. 11
Influenza and Respiratory Viruses, Bureau of
Microbiology, Laboratory Center for Disease Control, Federal
Laboratories, Winnipeg, Manitoba R3M 3R2, Canada
Received 28 April 1998/Returned for modification 26 June
1998/Accepted 18 August 1998
Diagnosis of respiratory virus infections currently involves
detection by isolation or antigen detection, which usually identifies only a single suspected agent. To permit identification of more than
one respiratory virus in clinical specimens, a rapid detection method
involving a single-step, multiplex reverse transcription-PCR (RT-PCR)
assay was developed. The assay included five primer sets that amplified
the RNA of respiratory syncytial virus subtypes A and B, parainfluenza
virus types 1, 2, and 3, and adenovirus types 1 to 7. Initially the
assay was tested on tissue culture-grown virus and was found to be
specific for all 12 prototype viruses tested, with no interassay cross
amplification or amplification of other respiratory viruses. Assay
sensitivity allowed a detection range of 0.2 50% tissue culture
infectious dose (TCID50) for adenovirus to 250 TCID50 for parainfluenza virus type 1. The multiplex RT-PCR assay was also able to directly detect viruses in respiratory specimens, with virus being detected in 41 of 112 samples as compared to 34 of 112 samples detected by direct immunofluorescence or antigen
detection following specimen culture. This suggests that the multiplex
RT-PCR assay can be used as a rapid and sensitive diagnostic method for
major respiratory viruses.
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Direct Detection of Respiratory Syncytial Virus, Parainfluenza
Virus, and Adenovirus in Clinical Respiratory Specimens by a
Multiplex Reverse Transcription-PCR Assay
*
Present address: Bloodborne Pathogens and Hepatitis,
Bureau of Microbiology, Laboratory Center for Disease Control, Federal Laboratories, 1015 Arlington St., Winnipeg, Manitoba R3M 3R2, Canada.
Phone: (204) 789-6061. Fax: (204) 789-2082. E-mail:
carla_osiowy{at}hc-sc.gc.ca.
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