Rapid and sensitive detection of mumps virus RNA directly from clinical samples by real-time PCR.
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.
Uchida
Kazue
K
Division of Virology, Saitama Institute of Public Health, Kamiökubo, Sakura-ku, Saitama City, Saitama Prefecture, Japan. a0183437@perf.saitama.lg.jp
Shinohara
Michiyo
M
Shimada
Shin-ichi
S
Segawa
Yukari
Y
Doi
Rie
R
Gotoh
Atushi
A
Hondo
Ryo
R
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
D012367:RNA, Viral; D014760:Viral Fusion Proteins