Loop-mediated isothermal amplification-based diagnostic assay for monkeypox virus infections
Article first published online: 20 APR 2009
Copyright © 2009 Wiley-Liss, Inc.
Journal of Medical Virology
Volume 81, Issue 6, pages 1102–1108, June 2009
How to Cite
Iizuka, I., Saijo, M., Shiota, T., Ami, Y., Suzaki, Y., Nagata, N., Hasegawa, H., Sakai, K., Fukushi, S., Mizutani, T., Ogata, M., Nakauchi, M., Kurane, I., Mizuguchi, M. and Morikawa, S. (2009), Loop-mediated isothermal amplification-based diagnostic assay for monkeypox virus infections. J. Med. Virol., 81: 1102–1108. doi: 10.1002/jmv.21494
- Issue published online: 20 APR 2009
- Article first published online: 20 APR 2009
- Manuscript Accepted: 13 MAR 2009
- Ministry of Health, Labor, and Welfare, Japan. Grant Numbers: H19-Shinko-Ippan-012, H19-Shinko-003
- loop-mediated isothermal amplification;
- monkeypox virus;
Monkeypox virus (MPXV) causes a smallpox-like disease in non-human primates and humans. This infection is endemic to central and western Africa. MPXV is divided into two genetically different groups, Congo Basin and West African MPXV, with the former being the more virulent. A real-time quantitative MPXV genome amplification system was developed for the diagnosis of MPXV infections using loop-mediated isothermal amplification (LAMP) technology. Primers used for genome amplification of Congo Basin (C-LAMP), West African (W-LAMP), and both Congo Basin and West African (COM-LAMP) MPXV by LAMP were designed according to the nucleotide sequences of the Congo Basin-specific D14L gene, the West African-specific partial ATI gene, and the partial ATI gene that is shared by both groups, respectively. The sensitivity and specificity of the LAMP were evaluated with nested PCR using peripheral blood and throat swab specimens collected from Congo Basin MPXV or West African MPXV-infected monkeys. The sensitivity and specificity of COM-LAMP, C-LAMP, and W-LAMP were 80% (45/56) and 100% (64/64); 79% (19/24) and 100% (24/24); and 72% (23/32) and 100% (40/40), respectively. The viremia level determined by LAMP assays increased with increases in the severity of the monkeypox-associated symptoms. The newly developed LAMP assay was confirmed to be a rapid, quantifiable, and highly sensitive and specific system effective in the diagnosis of MPXV infections. The LAMP assays made it possible to discriminate between Congo Basin and West African MPXV. The LAMP developed in this study is useful not only for diagnosis of but also for the assessment of MPXV infections. J. Med. Virol. 81:1102–1108, 2009. © 2009 Wiley-Liss, Inc.