2005 Fiscal Year Final Research Report Summary
Development of a rapid molecular diagnostic method for malaria at endemic villages
| Project/Area Number |
16590341
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Parasitology (including Sanitary zoology)
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| Research Institution | OITA UNIVERSITY |
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Principal Investigator |
KAWAMOTO Fumihiko OITA UNIVERSITY, Inst.Scientific Research, Prof., 総合科学研究支援センター, 教授 (40115556)
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| Co-Investigator(Kenkyū-buntansha) |
KANBE Toshio Nagoya Univ., Grad.Sch., Lecturer, 大学院・医学系研究科, 講師 (50093018)
OTSUKA Yasushi OITA UNIVERSITY, Faculty of Medicine, Research Assoc., 医学部, 助手 (00244161)
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| Project Period (FY) |
2004 – 2005
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| Keywords | Malaria / Molecular diagnosis / Rapid diagnosis / Simple diagnosis / Lamp method / Topo-isomerase II / Gene analysis / Rapid DNA extraction |
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| Research Abstract |
Molecular diagnosis for malaria by Nested PCR method is highly sensitive, but it is impossible to apply at the fields of malaria endemic areas. Recently, a new molecular amplification method (Lamp method) was reported. This method is simple and DNA is easily amplified only by incubating reagents with at 63 C. Furthermore, amplified product can be detected by naked eyes without any equipment. Therefore, this study was aimed to develop a new molecular diagnosis for malaria using the Lamp method.
Primer sets were designed from a target sequence in 18S ribosomal RNA gene. Using this set, DNA of malaria parasites was amplified greatly and white precipitates was seen in the bottom of the reaction tubes within 30 min to 1 hrs, indicating that malaria diagnosis was possible by this method. Application of this method in a field at Flores island, Indonesia, was also successful and detection lebel was the same with a nested PCR diagnosis.
For speciation of the four malaria species, it was impossible to design good primer sets from the 18S RNA gene because of too short target sequences. In order to use Topoisomerase II gene which has been used for speciation of yeasts and Candida, we have been analyzing its gene in malaria parasites and design primer sets for the speciation of the human malaria parasites.
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