This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Okafuji, T.
Right arrow Articles by Nakayama, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Okafuji, T.
Right arrow Articles by Nakayama, T.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, April 2005, p. 1625-1631, Vol. 43, No. 4
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.4.1625-1631.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Rapid Diagnostic Method for Detection of Mumps Virus Genome by Loop-Mediated Isothermal Amplification

Takao Okafuji,1,2 Naoko Yoshida,2 Motoko Fujino,2 Yoshie Motegi,2 Toshiaki Ihara,3 Yoshinori Ota,4 Tsugunori Notomi,4 and Tetsuo Nakayama2*

Okafuji Pediatric Clinic, Himeji, Hyougo,1 Kitasato Institute for Life Sciences, Laboratory of Viral Infection I, Minato-ku,2 Eiken Chemical Co. Ltd., Kita-ku, Tokyo,4 Department of Pediatrics, Mie National Hospital, Tsu, Mie, Japan3

Received 6 July 2004/ Returned for modification 27 August 2004/ Accepted 23 November 2004

Most mumps patients are clinically diagnosed without any virological examinations, but some diagnosed cases of mumps may be caused by other pathogens or secondary vaccine failure (SVF). To clarify these issues, a sensitive, specific, and rapid diagnostic method is required. We obtained 60 salivary swabs from 34 patients with natural infection during the course of the illness, 10 samples from patients with vaccine-associated parotitis, and 5 samples from patients with SVF. Total RNA was extracted and subjected to reverse transcription-PCR (RT-PCR) and loop-mediated isothermal amplification (LAMP) for genome amplification. We detected mumps virus RNA corresponding to 0.1 PFU by LAMP within 60 min after RNA extraction, with the same sensitivity as RT-nested PCR. Mumps virus was isolated in 30 of 33 samples within day 2, and mumps virus genome was amplified by LAMP in 32 of them. The quantity of virus titer was calculated by monitoring the time to reach the threshold of turbidity. The viral load decreased after day 3 and was lower in patients serologically diagnosed as having SVF with milder illness. Accuracy of LAMP for the detection of mumps virus genome was confirmed; furthermore, it is of benefit for calculating the viral load, which reflects disease pathogenesis.

* Corresponding author. Mailing address: Laboratory of Viral Infection I, Kitasato Institute for Life Sciences, Shirokane 5-9-1, Minato-ku, Tokyo 108-8641, Japan. Phone: 81-3-5791-6269. Fax: 81-3-5791-6130. E-mail: tetsuo-n{at}lisci.kitasato-u.ac.jp.

Journal of Clinical Microbiology, April 2005, p. 1625-1631, Vol. 43, No. 4
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.4.1625-1631.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Boddicker, J. D., Rota, P. A., Kreman, T., Wangeman, A., Lowe, L., Hummel, K. B., Thompson, R., Bellini, W. J., Pentella, M., DesJardin, L. E. (2007). Real-Time Reverse Transcription-PCR Assay for Detection of Mumps Virus RNA in Clinical Specimens. J. Clin. Microbiol. 45: 2902-2908 [Abstract] [Full Text]  
  • Mori, N., Motegi, Y., Shimamura, Y., Ezaki, T., Natsumeda, T., Yonekawa, T., Ota, Y., Notomi, T., Nakayama, T. (2006). Development of a new method for diagnosis of rubella virus infection by reverse transcription-loop-mediated isothermal amplification.. J. Clin. Microbiol. 44: 3268-3273 [Abstract] [Full Text]  
  • Fukuda, S., Takao, S., Kuwayama, M., Shimazu, Y., Miyazaki, K. (2006). Rapid Detection of Norovirus from Fecal Specimens by Real-Time Reverse Transcription-Loop-Mediated Isothermal Amplification Assay. J. Clin. Microbiol. 44: 1376-1381 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»