Grant-in-Aid for international Scientific Research
|Allocation Type||Single-year Grants |
|Research Institution||University of Shizuoka |
YANAGIHARA Yasutake University of Shizuoka, Professor, 薬学部, 教授 (30046255)
JOHNSON Russ ミネソタ大学, 医学部(米国), 教授
FUKUNAGA Masahito Fukuyama University, Professor, 薬学部, 教授 (20132483)
MASUZAWA Toshiyuki University of Shizuoka, Research Associate, 薬学部, 助手 (10181645)
IWAMOTO Yoshihisa University of Shizuoka, Associate Professor, 薬学部, 助教授 (60046290)
JOHNSON Russell C. University of Minnesota, U.S.A., Professor
R C.Johnson 米国ミネソタ大学, 医学部, 教授
|Project Period (FY)
1992 – 1993
Completed(Fiscal Year 1993)
|Budget Amount *help
¥3,900,000 (Direct Cost : ¥3,900,000)
Fiscal Year 1993 : ¥1,900,000 (Direct Cost : ¥1,900,000)
Fiscal Year 1992 : ¥2,000,000 (Direct Cost : ¥2,000,000)
|Keywords||Lyme disease / Lyme disease borreliae / PCR / Serodiagnosis / Antibiotics / Vaccine / Genediagnosis / Borrelia japonica / ライム菌 / FRLP / rRNA遺伝子 / 鞭毛蛋白遺伝子 / シュルツェマダニ / ヤマトマダニ|
Dr.Johnson was invited to Japan and he gave lectures on present status of Lyme borreliosis research at University of Shizuoka and National Institute of Health of Japan. Masuzawa was dispatched to Dr.Johnson's laboratory, WHO work-shop on Lyme borreliosis control at International Zoonoses Conference, WHO Head-quarters, University of Neuchatel and Pasteur Institute to discuss on Lyme disease and borreliae.
RFLP ribotyping of 23S rRNA gene of isolates from the world corresponded to 3 genospecies, Borrelia burgdorferi, B. garinii and B. afzelii, and were applicable to classification of genospecies of borreliae. Genomic Southern hybridization of borrelial isolates with 23S rRNA probes showed that borreliae originated from I. persulcatus were classified into B. garinii, B. afzelii and others.
Borrelia isolated from I. ovatus in Japan was named B. japonica as new species based on morphology, DNA homology, G+C content, RFLP analysis of 16S rRNA genes and reactivity to monoclonal antibodies. Mono
clonal antibodies capable of classifying B. japonica and other borreliae were prepared.
RFLP analysis using 16S rRNA gene probe BBU30 realized classification and identification of 4 genospecies and revealed that borreliae isolated from I. persulcatus and Apodemus speciosus ainu were belonging to B. garinii.
By PCR targetting flagellin gene, specifically amplified DNAs were detected from culture and experimentally infected ticks and mice. Two steps PCR detected 1 Borrelia cell and is applicable to gene diagnosis and epidemiological study.
PCR for Osp A and Osp B and RFLP analysis of the PCR products were useful for direct identification of 4 genospecies from clinical samples.
Specific lesion was induced by inoculation of pathogenic borreliae into footpad of outbred ddY mice. This animal model was useful in research for developingvaccine. Cross protection was not found among strains having different antigens of Osp A and B. Furthermore, Borrelia strains corresponding to endemic area are necessary for vaccine. The PCR developed in this study are useful for rapid identification of endemic strain. Less