| Co-Investigator(Kenkyū-buntansha) |
KIMURA Mikio National Institute of Infectious disease, Head of office, 感染症情報センター, 室長 (90114462)
KIM Hye-sook Okayama University, The Graduate School of Natural Sciences and Technology, Assistant Professor, 大学院・自然科学研究科, 講師 (70314664)
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| Research Abstract |
We have developed a colorimetric assay, "microtiter plate-hybridization", for the detection of malaria parasites in human blood without phenol extraction of DNA, in which the target DNA sequences (18S small subunit ribosomal RNA gene) amplified by polymerase chain reaction (PCR) are hybridized with the species-specific probes immobilized on a microtiter well. The PCR products bound on a well are visualized by the biotin-streptavidin system and the following chromogenic reaction in a manner similar to that of an enzyme immunoassay. This assay system allowed us to detect and identify the four species of human malaria parasites (Plasmodium falciparum, P. vivax. P. ovale and P. malariae) and one variant of P. ovale, each parasite-specific microtiter plates were tested for imported malaria in Japan. We obtained blood samples by finger puncture from 327 asymptomatic donors, almost they had medical examination at the Institute of Medical Science, The University of Tokyo. Among the 327 samples, 118 (36.1%) were P. falciparum-positive, 83 (25.4%) were P. vivax-positive, 16 (4.9%) were P. ovale-positive, 3 (0.9%) were P. malariae-positive and negative were 101 cases. Our method provides a rapid and sensitive assay, and thus may be useful for large scale screening or assessment of malaria control programs in endemic areas.
Furthermore, we developed of quick diagnostic system of human malaria parasites for the use of malaria endemic areas, loop-mediated isothermal amplification (LAMP), this method is simple, PCR equipment is not required and detection time is about 30 minutes. We are developing the LAMP for detection of malaria parasites in endemic areas.
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