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

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, September 2006, p. 3268-3273, Vol. 44, No. 9
0095-1137/06/$08.00+0     doi:10.1128/JCM.00803-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Development of a New Method for Diagnosis of Rubella Virus Infection by Reverse Transcription-Loop-Mediated Isothermal Amplification

Kitasato Institute for Life Sciences, Laboratory of Viral Infection I, Minato-ku, Tokyo 108-8641, Japan,¹ Ashikaga Red Cross Hospital, Department of Pediatrics, Ashikaga, Tochigi 326-0808, Japan,² Eiken Chemical Co. Ltd., Kita-ku, Tokyo 114-0002, Japan,³ Kansai Medical University, Department of Pediatrics, Moriguchi, Osaka 570-8506, Japan⁴

Received 21 December 2005/ Returned for modification 11 June 2006/ Accepted 9 July 2006

We developed a useful method for the detection of rubella virus genome RNA by reverse transcription loop-mediated isothermal amplification (RT-LAMP) and compared the sensitivity of RT-LAMP with that of other virological tests: reverse transcription-PCR (RT-PCR) and virus isolation. The rubella virus genome was amplified by RT-LAMP from clinical isolates obtained between 1987 and 2004 with similar sensitivities to the Takahashi vaccine strain. The detection limit of RT-LAMP was compared with that of RT-PCR using the Takahashi vaccine strain. We detected rubella virus genome material corresponding to 30 PFU/ml in a culture fluid sample by RT-LAMP within 60 min after the extraction of RNA with equal sensitivity to RT-nested PCR. The positive result rates of RT-LAMP, RT-PCR, and virus isolation were also compared using throat swabs obtained from patients who were clinically diagnosed with acute rubella virus infection in 2004 in Tochigi, Japan. Among nine patients with clinical rubella, the positive result rates were three/nine (33.3%) for virus isolation, six/nine (66.7%) for RT-PCR, and seven/nine (77.8%) for RT-LAMP. Consequently, RT-LAMP for rubella virus would be expected to be a reliable rapid diagnostic tool in the clinical setting.