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Journal of Clinical Microbiology, October 1998, p. 2990-2995, Vol. 36, No. 10
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Multiplex PCR for Typing and Subtyping Influenza and Respiratory Syncytial Viruses

J. Stockton,1 J. S. Ellis,1 M. Saville,2 J. P. Clewley,1 and M. C. Zambon1,*

Virus Reference Division, Central Public Health Laboratory, Public Health Laboratory Service, London NW9 5HT,1 and Department of Diagnostic Virology, St. Marys Hospital, Paddington, London W2,2 United Kingdom

Received 18 February 1998/Returned for modification 29 April 1998/Accepted 1 July 1998

A multiplex reverse transcription (RT)-PCR method that has been developed is capable of detecting and subtyping influenza A (H1N1 and H3N2) and B viruses as well as respiratory syncytial virus (RSV) types A and B in respiratory clinical samples taken as part of a national community-based surveillance program of influenza-like illness in England and Wales. The detection of each different pathogen depended on distinguishing five amplification products of different sizes on agarose gels following RT-PCR with multiple primer sets. The multiplex RT-PCR was tested with 65 nasopharyngeal apirates from which RSV had been isolated and 237 combined nose and throat swabs from which influenza A (H1N1 and H3N2) or B virus had been detected by virus isolation, as well as 40 respiratory samples from which other viruses including cytomegalovirus, herpes simplex virus, enteroviruses, and parainfluenza viruses had been grown. For the typing and subtyping of influenza A and B viruses and RSV types A and B, the multiplex RT-PCR gave an excellent (100%) correlation with the results of conventional typing and subtyping with specific antisera. Multiplex RT-PCR can also be used to accurately detect more than one viral template in the same reaction mixture, allowing viral coinfections to be identified with the same respiratory specimen.

* Corresponding author. Mailing address: Central Public Health Laboratory, 61 Colindale Ave., London NW9 5HT, United Kingdom. Phone: 0181 200 4400. Fax: 0181 200 1569. E-mail: mzambon{at}phls.co.uk.

Journal of Clinical Microbiology, October 1998, p. 2990-2995, Vol. 36, No. 10
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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