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Am. J. Trop. Med. Hyg., 81(4), 2009, pp. 578-582
doi:10.4269/ajtmh.2009.09-0145;
Copyright © 2009 by The American Society of Tropical Medicine and Hygiene

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Sensitive, Specific, and Rapid Detection of Leishmania donovani DNA by Loop-Mediated Isothermal Amplification

Hidekazu Takagi, Makoto Itoh*, Mohammad Zahidul Islam, Abdur Razzaque, A. R. M. Saifuddin Ekram, Yoshihisa Hashighuchi, Eisei Noiri, AND Eisaku Kimura
Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan; Department of Medicine, Rajshahi Medical College Hospital, Rajshahi, Bangladesh; Department of Parasitology, Kochi Medical School, Kochi University, Nankoku, Kochi, Japan; Department of Nephrology and Endocrinology, and Department of Hemodialysis and Apheresis, University Hospital, The University of Tokyo, Tokyo, Japan

We have applied a loop-mediated isothermal amplification (LAMP) technique to detect Leishmania donovani DNA. The LAMP technique detected 1 fg of L. donovani DNA, which was 10-fold more sensitive than a conventional polymerase chain reaction (PCR). All nested PCR–positive blood samples from visceral leishmaniasis patients were positive with the LAMP technique, and DNA samples from L. infantum, L. major, L. mexicana, L. tropica, L. braziliensis, Plasmodium falciparum, and healthy humans were negative with the LAMP technique. The advantages of the LAMP method are its shorter reaction time, a lack of requirement of sophisticated equipment, and visual judgment of positivity based on the turbidity of reaction mixture. Our LAMP technique can be a better alternative to a conventional PCR, especially under field conditions.

Received March 18, 2009. Accepted for publication June 17, 2009.

Financial support: This study was supported by the Special Coordination Funds for Promoting Science and Technology of MEXT of Japan, 1200015, and also by Grants-in-Aid for Scientific Research (B) No. 18406013 from the Japan Society for the Promotion of Science.

* Address correspondence to MakotoItoh, Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi-gun, 480-1195, Japan. E-mail: macitoh{at}aichi-med-u.ac.jp

Authors’ addresses: Hidekazu Takagi, Makoto Itoh, Mohammad Zahidul Islam, and Eisaku Kimura, Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan. Abdur Razzaque and A. R. M. Saifuddin Ekram, Department of Medicine, Rajshahi Medical College Hospital, Rajshahi, Bangladesh. Yoshihisa Hashighuchi, Department of Parasitology, Kochi Medical School, Kochi University, Nankoku, Kochi, Japan. Eisei Noiri, Department of Nephrology and Endocrinology, and Department of Hemodialysis and Apheresis, University Hospital, The University of Tokyo, Tokyo, Japan.






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Copyright © 2009 by the American Society of Tropical Medicine and Hygiene.