Research Article

Loop-mediated isothermal amplification-based diagnostic assay for monkeypox virus infections

  1. Itoe Iizuka1,2,
  2. Masayuki Saijo1,*,
  3. Tomoyuki Shiota1,2,
  4. Yasushi Ami3,
  5. Yuriko Suzaki3,
  6. Noriyo Nagata4,
  7. Hideki Hasegawa4,
  8. Kouji Sakai1,3,
  9. Shuetsu Fukushi1,
  10. Tetsuya Mizutani1,
  11. Momoko Ogata1,
  12. Mina Nakauchi1,
  13. Ichiro Kurane1,
  14. Masashi Mizuguchi2,
  15. Shigeru Morikawa1

Article first published online: 20 APR 2009

DOI: 10.1002/jmv.21494

Issue

Journal of Medical Virology

Journal of Medical Virology

Volume 81, Issue 6, pages 1102–1108, June 2009

Additional Information

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. Journal of Medical Virology, 81: 1102–1108. doi: 10.1002/jmv.21494

Author Information

  1. 1

    Department of Virology 1, National Institute of Infectious Diseases, Musashimurayama, Tokyo, Japan

  2. 2

    Faculty of Medicine, Department of International Health, The University of Tokyo, Bunkyo-ku, Tokyo, Japan

  3. 3

    Department of Experimental Animals, National Institute of Infectious Diseases, Musashimurayama, Tokyo, Japan

  4. 4

    Department of Pathology, National Institute of Infectious Diseases, Musashimurayama, Tokyo, Japan

*Department of Virology 1, National Institute of Infectious Disease, Tokyo 208-0011, Japan.

Publication History

  1. Issue published online: 20 APR 2009
  2. Article first published online: 20 APR 2009
  3. Manuscript Accepted: 13 MAR 2009

Funded by

  • Ministry of Health, Labor, and Welfare, Japan. Grant Numbers: H19-Shinko-Ippan-012, H19-Shinko-003

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Keywords:

  • loop-mediated isothermal amplification;
  • LAMP;
  • monkeypox;
  • monkeypox virus;
  • diagnosis

Abstract

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.

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