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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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