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

Masaru Ihira, Tetsushi Yoshikawa, Yoshihiko Enomoto, Shiho Akimoto, Masahiro Ohashi, Sadao Suga, Naoko Nishimura, Takao Ozaki, Yukihiro Nishiyama, Tsugunori Notomi, Yoshinori Ohta, Yoshizo Asano

Research output: Contribution to journalArticle

80 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)140-145
Number of pages6
JournalJournal of clinical microbiology
Volume42
Issue number1
DOIs
Publication statusPublished - 01-01-2004

    Fingerprint

All Science Journal Classification (ASJC) codes

  • Microbiology (medical)

Cite this

Ihira, M., Yoshikawa, T., Enomoto, Y., Akimoto, S., Ohashi, M., Suga, S., Nishimura, N., Ozaki, T., Nishiyama, Y., Notomi, T., Ohta, Y., & Asano, Y. (2004). Rapid Diagnosis of Human Herpesvirus 6 Infection by a Novel DNA Amplification Method, Loop-Mediated Isothermal Amplification. Journal of clinical microbiology, 42(1), 140-145. https://doi.org/10.1128/JCM.42.1.140-145.2004