title: Rapid and sensitive detection of mumps virus RNA directly from clinical samples by real-time PCR.
abstract: 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.
authors.affiliation: Division of Virology, Saitama Institute of Public Health, Kamiökubo, Sakura-ku, Saitama City, Saitama Prefecture, Japan. a0183437@perf.saitama.lg.jp
authors.forename: Kazue
authors.initials: K
authors.lastname: Uchida
authors.forename: Michiyo
authors.initials: M
authors.lastname: Shinohara
authors.forename: Shin-ichi
authors.initials: S
authors.lastname: Shimada
authors.forename: Yukari
authors.initials: Y
authors.lastname: Segawa
authors.forename: Rie
authors.initials: R
authors.lastname: Doi
authors.forename: Atushi
authors.initials: A
authors.lastname: Gotoh
authors.forename: Ryo
authors.initials: R
authors.lastname: Hondo
mesh_terms: D006801:Humans; D008582:Meningitis, Aseptic / Q000821:virology; D009107:Mumps / Q000821:virology; D009109:Mumps virus / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014760:Viral Fusion Proteins / Q000235:genetics; D014773:Virology / Q000379:methods
chemical_list: D012367:RNA, Viral; D014760:Viral Fusion Proteins