Open AccessHeminested Multiplex Reverse Transcription-PCR for Detection and Differentiation of Norwalk-Like Virus Genogroups 1 and 2 in Fecal Samples
Author(s) -
Lilly Yuen,
Michael Catton,
Bradley J Cox,
P.J. Wright,
John A. Marshall
Publication year - 2001
Publication title -
journal of clinical microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.349
H-Index - 255
eISSN - 1070-633X
pISSN - 0095-1137
DOI - 10.1128/jcm.39.7.2690-2694.2001
Subject(s) - multiplex , norwalk virus , feces , biology , virology , reverse transcription polymerase chain reaction , multiplex polymerase chain reaction , caliciviridae , reverse transcriptase , polymerase chain reaction , virus , genetics , microbiology and biotechnology , norovirus , gene , messenger rna
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 microl 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.