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

doi:10.1128/JCM.39.8.2779-2783.2001

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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.) wasused for the simultaneous detection of human parainfluenza virustypes 1, 2, and 3, influenza virus types A and B, and respiratorysyncytial virus types A and B. One hundred forty-three respiratoryspecimens from 126 patients were analyzed by RT-PCR-EHA, and theresults were compared to those obtained by conventional viralculture and immunofluorescence (IF) methods. RT-PCR-EHA provedto be positive for 17 of 143 (11.9%) specimens, whereas 8 of 143(5.6%) samples were positive by viral culture and/or IF. Eightsamples were positive by both RT-PCR-EHA and conventional methods,while nine samples were RT-PCR-EHA positive and viral cultureand IF negative. Eight of the nine samples with discordant resultswere then independently tested by a different multiplex RT-PCRassay for influenza virus types A and B, and all eight provedto 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 positivesamples, which would otherwise have been missed by routine methods,we suggest that this multiplex RT-PCR-EHA provides a highly sensitiveand specific means of diagnostic detection of major respiratoryviruses.

^* 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.

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