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Rapid and sensitive detection of mumps virus RNA directly from clinical samples by real‐time PCR
Author(s) -
Uchida Kazue,
Shinohara Michiyo,
Shimada Shinichi,
Segawa Yukari,
Doi Rie,
Gotoh Atushi,
Hondo Ryo
Publication year - 2005
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.20291
Subject(s) - mumps virus , virology , virus , real time polymerase chain reaction , aseptic meningitis , biology , meningitis , reverse transcription polymerase chain reaction , encephalitis , serial dilution , paramyxoviridae , viral culture , polymerase chain reaction , microbiology and biotechnology , medicine , viral disease , gene , gene expression , pathology , biochemistry , alternative medicine , psychiatry
A rapid, sensitive, and specific assay to detect mumps virus RNA directly from clinical specimens using a real‐time PCR assay was developed. The assay was capable of detecting five copies of standard plasmid containing cDNA from the mumps virus F gene. No cross‐reactions were observed with other members of Paramyxoviridae , or with viruses or bacteria known to be meningitis pathogens. Seventy‐three clinical samples consisting of throat swabs collected from patients with parotitis, and cerebrospinal fluid (CSF) collected from patients with aseptic meningitis, were examined with a real‐time PCR assay developed by the authors, reverse‐transcription nested‐PCR (RT‐n‐PCR), and virus isolation using cell culture. Like the RT‐n‐PCR assay, the real‐time PCR assay could detect mumps virus RNA in approximately 70% of both throat swabs and CSF samples, while, by tissue culture, mumps virus was isolated from only approximately 20% of CSF and 50% of throat swab samples. In addition, the real‐time PCR assay could be developed easily into a quantitative assay for clinical specimens containing more than 1,800 copies of mumps virus RNA/ml by using serial dilutions of the standard plasmid. The results suggest that the real‐time PCR assay is useful for identification of mumps virus infections, not only in typical cases, but also in suspected cases, which show only symptoms of meningitis or encephalitis. J. Med. Virol. 75:470–474, 2005. © 2005 Wiley‐Liss, Inc.