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Detection of Trypanosoma cruzi and T. rangeli Infections from Rhodnius pallescens Bugs by Loop-Mediated Isothermal Amplification (LAMP)

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  • National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Japan; College of Medicine, Department of Microbiology, University of Panamá, Panama; Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan

We have developed two loop-mediated isothermal amplification (LAMP) assays for specific detection of Trypanosoma cruzi and Trypanosoma rangeli based on the 18S ribosomal RNA (rRNA) and the small nucleolar RNA (snoRNA) genes, respectively. The detection limit of the assays is 100 fg and 1 pg for T. cruzi and T. rangeli, respectively, with reactions conducted in 60 minutes. The two LAMP assays were used in detection of T. cruzi and T. rangeli infections in comparison with polymerase chain reaction (PCR) for DNA samples extracted from Rhodnius pallescens bugs collected from palm trees in Panama. Out of a total of 52 DNA samples from R. pallescens bugs 17 (33%) and 14 (27%) were T. cruzi-positive by LAMP and PCR, respectively, while, 7 (13%) and 4 (8%) were T. rangeli-positive by LAMP and PCR, respectively. Further evaluation of these LAMP assays is needed, especially with specimens collected from human patients as well as blood kept for transfusion purposes.

Author Notes

*Address correspondence to Noboru Inoue, National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan. E-mail: ircpmi@obihiro.ac.jp

Financial support: The first author was supported by a research grant fellowship for young scientists from the Japanese Society for the Promotion of Science (JSPS). FR and AMC-S received training on protozoan diseases at NRCPD as Japan International Cooperation Agency (JICA) participants. This study was also made possible by a Grant-in-aid for scientific research from JSPS to NI.

Authors' addresses: Oriel M. M. Thekisoe, Andrea M. Coronel-Servian, Shinya Fukumoto, Shin-Ichiro Kawazu, and Noboru Inoue, National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan, E-mails: thekisoe@yahoo.com, andycoronel@yahoo.com, fukumoto@obihiro.ac.jp, skawazu@obihiro.ac.jp, and ircpmi@obihiro.ac.jp. Carol V. Rodriguez and Francisco Rivas, College of Medicine, Department of Microbiology, University of Panamá, Panama, E-mails: carolrodriguez_24@hotmail.com and frivast@hotmail.com. Chihiro Sugimoto, Research Center for Zoonosis Control, Hokkaido University, Sapporo, Hokkaido, Japan, E-mail: sugimoto@czc.hokudai.ac.jp.

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