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Molecular Biology of Lyme Borreliosis and Lyme Disease Control

莱姆疏螺旋体病的分子生物学和莱姆病控制

基本信息

批准号：
04044138
负责人：
YANAGIHARA Yasutake
金额：
$ 2.5万
依托单位：
University of Shizuoka
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1993
项目状态：
已结题

项目摘要

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 … More 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

johnson博士应邀赴日本静冈大学和日本国立卫生研究院就莱姆病研究现状进行了讲座。Masuzawa被派往johnson博士的实验室、世卫组织在国际人兽共患病会议上控制莱姆病和疏螺旋体病的车间、世卫组织总部、纳沙泰尔大学和巴斯德研究所，讨论莱姆病和疏螺旋体问题。全球分离株23S rRNA基因的RFLP分型分别对应伯氏疏螺旋体、B. garinii和B. afzelii 3个基因种，可用于伯氏疏螺旋体的基因种分类。利用23S rRNA探针对borreliae分离株进行基因组Southern杂交，结果表明，源自persulcatus的borreliae可分为B. garinii、B. afzelii等。根据形态、DNA同源性、G+C含量、16S rRNA基因的RFLP分析和对单克隆抗体的反应性，将日本卵形伊蚊分离的疏螺旋体命名为新种。制备了更多的单克隆抗体，能够对日本疏螺旋体和其他疏螺旋体进行分类。采用16S rRNA基因探针BBU30进行RFLP分析，实现了4个基因种的分类和鉴定，结果表明，从过槽伊蚊和阿努斑姬蚊中分离到的螺旋体属加里尼布氏螺旋体。采用PCR方法，从培养和实验感染的蜱和小鼠中检测到鞭毛蛋白基因特异性扩增的dna。两步PCR检测出1个伯氏疏螺旋体细胞，适用于基因诊断和流行病学研究。对Osp A和Osp B的PCR和PCR产物的RFLP分析可用于临床样品中4个基因种的直接鉴定。用致病性疏螺旋体接种远交系小鼠足垫，引起特异性病变。该动物模型对研制疫苗有一定的参考价值。具有不同抗原的疏螺旋体A、b株间未发现交叉保护，与疫区相对应的疏螺旋体菌株需要接种疫苗。本研究建立的PCR方法可用于快速鉴定地方性毒株。少