Real-time reverse transcription-PCR assay for detection of mumps virus RNA in clinical specimens.
The mumps virus is a negative-strand RNA virus in the family Paramyxoviridae. Mumps infection results in an acute illness with symptoms including fever, headache, and myalgia, followed by swelling of the salivary glands. Complications of mumps can include meningitis, deafness, pancreatitis, orchitis, and first-trimester abortion. Laboratory confirmation of mumps infection can be made by the detection of immunoglobulin M-specific antibodies to mumps virus in acute-phase serum samples, the isolation of mumps virus in cell culture, or by detection of the RNA of the mumps virus by reverse transcription (RT)-PCR. We developed and validated a multiplex real-time RT-PCR assay for rapid mumps diagnosis in a clinical setting. This assay used oligonucleotide primers and a TaqMan probe targeting the mumps SH gene, as well as primers and a probe that targeted the human RNase P gene to assess the presence of PCR inhibitors and as a measure of specimen quality. The test was specific, since it did not amplify a product from near-neighbor viruses, as well as sensitive and accurate. Real-time RT-PCR results showed 100% correlation with results from viral culture, the gold standard for mumps diagnostic testing. Assay efficiency was over 90% and displayed good precision after performing inter- and intraassay replicates. Thus, we have developed and validated a molecular method for rapidly diagnosing mumps infection that may be used to complement existing techniques.
Boddicker
Jennifer D
JD
University of Iowa Hygienic Laboratory, University of Iowa, 102 Oakdale Hall, H101-OH, Iowa City, IA 52242, USA.
Rota
Paul A
PA
Kreman
Trisha
T
Wangeman
Andrea
A
Lowe
Louis
L
Hummel
Kimberly B
KB
Thompson
Robert
R
Bellini
William J
WJ
Pentella
Michael
M
Desjardin
Lucy E
LE
D017931:DNA Primers / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009107:Mumps / Q000175:diagnosis / Q000821:virology*; D009109:Mumps virus / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000097:blood*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D043262:Ribonuclease P / Q000235:genetics; D012680:Sensitivity and Specificity; D013223:Statistics as Topic; D014764:Viral Proteins / Q000235:genetics; D014776:Virus Cultivation
D017931:DNA Primers; D012367:RNA, Viral; D014764:Viral Proteins; C067386:small hydrophobic protein, Mumps virus; D043262:Ribonuclease P