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Journal of Clinical Microbiology, July 2001, p. 2690-2694, Vol. 39, No. 7
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.7.2690-2694.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Heminested Multiplex Reverse Transcription-PCR for Detection and Differentiation of Norwalk-Like Virus Genogroups 1 and 2 in Fecal Samples

Lilly K. W. Yuen,1,* Michael G. Catton,1 Bradley J. Cox,1 Peter J. Wright,2 and John A. Marshall1

Victorian Infectious Diseases Reference Laboratory Carlton South, Victoria 3053,1 and Department of Microbiology, Monash University, Clayton, Victoria 3800,2 Australia

Received 6 November 2000/Returned for modification 3 February 2001/Accepted 6 May 2001

The present study describes a heminested multiplex reverse transcription (RT)-PCR assay which enables simultaneous detection and differentiation of Norwalk-like virus (NLV) genogroups from clinical fecal samples without the need to perform sequencing or hybridization. The assay developed was able to detect concentrations of fewer than 100 viral particles per 5 µl of clarified fecal extract and could differentiate the two genogroups with a specificity of 100%. Although the multiplex RT-PCR assay failed to detect NLV in about 3% of the fecal samples which were NLV positive by electron microscopy (EM), the assay was approximately six times more sensitive than EM for NLV detection.

* Corresponding author. Mailing address: VIDRL, 10 Wreckyn St., North Melbourne, 3051 Victoria, Australia. Phone: 61-3-9342-2619. Fax: 61-3-9342-2666. E-mail: lilly.yuen{at}mh.org.au.

Journal of Clinical Microbiology, July 2001, p. 2690-2694, Vol. 39, No. 7
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.7.2690-2694.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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