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Journal of Clinical Microbiology, September 2007, p. 2902-2908, Vol. 45, No. 9
0095-1137/07/$08.00+0     doi:10.1128/JCM.00614-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Real-Time Reverse Transcription-PCR Assay for Detection of Mumps Virus RNA in Clinical Specimens{triangledown}

Jennifer D. Boddicker,1 Paul A. Rota,2 Trisha Kreman,1 Andrea Wangeman,1 Louis Lowe,2 Kimberly B. Hummel,2 Robert Thompson,1 William J. Bellini,2 Michael Pentella,1 and Lucy E. DesJardin1*

University of Iowa Hygienic Laboratory, University of Iowa, Iowa City, Iowa,1 Measles, Mumps, Rubella, and Herpesviruses Laboratory Branch, Centers for Disease Control and Prevention, Atlanta, Georgia2

Received 20 March 2007/ Returned for modification 4 June 2007/ Accepted 16 July 2007

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.

* Corresponding author. Mailing address: University of Iowa Hygienic Laboratory, 102 Oakdale Hall, H101-OH, Iowa City, IA 52242. Phone: (319) 335-4339. Fax: (319) 335-4555. E-mail: ldesjard{at}uhl.uiowa.edu

{triangledown} Published ahead of print on 25 July 2007.

Journal of Clinical Microbiology, September 2007, p. 2902-2908, Vol. 45, No. 9
0095-1137/07/$08.00+0     doi:10.1128/JCM.00614-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Leblanc, J. J., Pettipas, J., Davidson, R. J., Tipples, G. A., Hiebert, J., Hatchette, T. F. (2008). Detection of Mumps Virus RNA by Real-Time One-Step Reverse Transcriptase PCR Using the LightCycler Platform. J. Clin. Microbiol. 46: 4049-4051 [Abstract] [Full Text]  
  • Bitsko, R. H., Cortese, M. M., Dayan, G. H., Rota, P. A., Lowe, L., Iversen, S. C., Bellini, W. J. (2008). Detection of RNA of Mumps Virus during an Outbreak in a Population with a High Level of Measles, Mumps, and Rubella Vaccine Coverage. J. Clin. Microbiol. 46: 1101-1103 [Abstract] [Full Text]  


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