| Project/Area Number |
01044021
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | Gunma Univrsity School of Medicine |
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Principal Investigator |
WAKI Seiji Gunma University School of Medicine Associate Professor, 医学部, 助教授 (10056286)
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| Co-Investigator(Kenkyū-buntansha) |
陳 林 中国第二軍医大学, 教授
顧 浩明 中国科学院, 上海葯物研究所, 副教授
GU Haoming Shanghhai Institute of Materia Medica, Chinese Academy of Sciences Associate Pro
CHEEN Lin The Second Military Medical College Professor
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| Project Period (FY) |
1989 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥7,200,000 (Direct Cost: ¥7,200,000)
Fiscal Year 1991: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1990: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1989: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | Malaria / Plasmodium falciparum / Drug resistance / Fluoroassay / DNA hybridization assay / Sero-epidemiology / Immunoglobulin isotype / DNAプロ-ブ / 感染防御免疫 |
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| Research Abstract |
The spread of drug-resistant Plasmodium falciparum accompanied by the world-wide resurgence of malaria urges the necessity, for the establishment of simplified and standardized method for in vitro drug sensitivity test. Currently drug sensitivity of the parasites has been examined by WHO standard method which requires visual differential counting of parasites by well-trained microscopists. We applied a fluoroassay using ethidium bromide to the test in order to enable objective reading of the results in the first year of the program. Reliable results were obtained in the test, however, the assay requires facilities such as a centrifuge and a fluorescent spectrophotometer that may not be available in the field. As the next step, a DNA hybridization technique was applied to the test using a synthetic oligonucleotide coupled to alkaline phosphatase. The density of the colored spot clearly distinguished development of parasites in culture. The new assay was assessed in Hainan Island, China, under field conditions. The trial demonstrated that, in the in vitro drug sensitivity test, the DNA hybridization technique could yield results parallel to those obtained by microscopical examination of blood smears. All procedures were carried out in an ordinary hospital laboratory on Hainan Island. Thus we consider that the new drug sensitivity test using a non-radioactive DNA probe is applicable to use in the field. We are developing some appropriate kits for the test and are expecting to be able to supply them to malaria endemic areas in the near future.
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