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Journal of Clinical Microbiology, August 2001, p. 2779-2783, Vol. 39, No. 8
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.8.2779-2783.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Comparison of a Multiplex Reverse Transcription-PCR-Enzyme Hybridization Assay with Conventional Viral Culture and Immunofluorescence Techniques for the Detection of Seven Viral Respiratory Pathogens

Lisa Liolios,1,* Adam Jenney,2 Denis Spelman,1,2 Tom Kotsimbos,3 Michael Catton,4 and Steve Wesselingh1

Infectious Diseases Unit1 and Departments of Microbiology2 and Respiratory Medicine,3 Alfred Hospital, Prahran 3181, and Victorian Infectious Diseases Reference Laboratory, North Melbourne 3051,4 Victoria, Australia

Received 8 January 2001/Returned for modification 18 March 2001/Accepted 13 May 2001

A multiplex reverse transcription-PCR-enzyme hybridization assay (RT-PCR-EHA; Hexaplex; Prodesse Inc., Waukesha, Wis.) was used for the simultaneous detection of human parainfluenza virus types 1, 2, and 3, influenza virus types A and B, and respiratory syncytial virus types A and B. One hundred forty-three respiratory specimens from 126 patients were analyzed by RT-PCR-EHA, and the results were compared to those obtained by conventional viral culture and immunofluorescence (IF) methods. RT-PCR-EHA proved to be positive for 17 of 143 (11.9%) specimens, whereas 8 of 143 (5.6%) samples were positive by viral culture and/or IF. Eight samples were positive by both RT-PCR-EHA and conventional methods, while nine samples were RT-PCR-EHA positive and viral culture and IF negative. Eight of the nine samples with discordant results were then independently tested by a different multiplex RT-PCR assay for influenza virus types A and B, and all eight proved to be positive. In comparison to viral culture and IF methods, RT-PCR-EHA gave a sensitivity and a specificity of 100 and 93%, respectively. Since RT-PCR-EHA was able to detect more positive samples, which would otherwise have been missed by routine methods, we suggest that this multiplex RT-PCR-EHA provides a highly sensitive and specific means of diagnostic detection of major respiratory viruses.

* Corresponding author. Mailing address: Infectious Diseases Unit, Alfred Hospital, Commercial Rd., Prahran, Victoria, Australia, 3181. Phone: 61-3-9276 3516. Fax: 61-3-9276 3424. E-mail: L.Liolios{at}alfred.org.au.

Journal of Clinical Microbiology, August 2001, p. 2779-2783, Vol. 39, No. 8
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.8.2779-2783.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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