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Journal of Clinical Microbiology, January 2004, p. 140-145, Vol. 42, No. 1
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.1.140-145.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Rapid Diagnosis of Human Herpesvirus 6 Infection by a Novel DNA Amplification Method, Loop-Mediated Isothermal Amplification

Masaru Ihira,1 Tetsushi Yoshikawa,2* Yoshihiko Enomoto,2 Shiho Akimoto,2 Masahiro Ohashi,2 Sadao Suga,2 Naoko Nishimura,3 Takao Ozaki,3 Yukihiro Nishiyama,4 Tsugunori Notomi,5 Yoshinori Ohta,5 and Yoshizo Asano2

Department of Medical Information Technology, Fujita Health University College,1 Department of Pediatrics, Fujita Health University School of Medicine, Toyoake,3 Department of Pediatrics, Showa Hospital, Konan, Aichi,2 Department of Virology, Nagoya University Graduate School of Medicine, Nagoya,4 Eiken Chemical Co., Ltd., Shimoishigami, Ohtawara, Tochigi, Japan5

Received 12 May 2003/ Returned for modification 4 July 2003/ Accepted 11 October 2003

A novel nucleic acid amplification method, termed loop-mediated isothermal amplification (LAMP), which amplifies DNA with high specificity, efficiency, and rapidity under isothermal conditions, may be a valuable tool for the rapid detection of infectious agents. LAMP was developed for human herpesvirus 6 (HHV-6), and its reliability was evaluated in this study. Although LAMP products were detected in HHV-6 B and HHV-6 A DNA, they were not detected in HHV-7 and human cytomegalovirus DNA. The sensitivity of the original HHV-6 LAMP protocol was 50 copies/tube. In order to increase the method's sensitivity, HHV-6 LAMP was modified by increasing the primer concentration. As a result of the modification, sensitivity increased to 25 copies/tube. After these initial validation studies, 13 patients with fever were tested for HHV-6 by viral isolation, serological analysis, and HHV-6 LAMP. In three of the eight patients with primary HHV-6 infection, HHV-6 DNA was detected in whole blood by the original HHV-6 LAMP protocol in not only the acute phase but also the convalescent phase. HHV-6 DNA was detected by modified HHV-6 LAMP in all eight plasma samples collected in the acute phase; however, no HHV-6 DNA was detected in plasma samples collected in the convalescent phase. Although HHV-6 DNA was detected in both the acute and convalescent phases of whole-blood samples in patients with past HHV-6 infection, it was not detected in plasma samples that did not contain latent viral DNA. Thus, detection of HHV-6 DNA in plasma by using this modified HHV-6 LAMP protocol is appropriate for diagnosis of active HHV-6 infection.

* Corresponding author. Mailing address: Department of Pediatrics, Fujita Health University School of Medicine, Toyoake 4701192, Japan. Phone: 81-562-939251. Fax: 81-562-952216. E-mail: tetsushi{at}fujita-hu.ac.jp.

Journal of Clinical Microbiology, January 2004, p. 140-145, Vol. 42, No. 1
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.1.140-145.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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