权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Development of a new epidemiological method using patterns of exoproteins secreted from bacteria

利用细菌分泌的外蛋白模式开发新的流行病学方法

基本信息

批准号：
12557027
负责人：
OHTA Michio
金额：
$ 7.87万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

项目摘要

Discrimination of each strain of bacteria such as Staphylococcus aureus and group A streptococcus (GAS) is helpful for tracing a pathogenic strain and prevention of nosocomial infections. Although DNA-based typings have been generally used as epidemiological methods for discrimination, we developed two dimensional electrophoretic (2-DE) analysis of exoproteins secreted from bacteria as a new epidemiological method and we found that this method was well applicable for the discrimination of S.aureus and GAS strains. The conditions of culture, growth and extraction of exoproteins were identified to have reproducible results. The major results of this study were as follows ; the best condition was that exoproteins were extracted from early stationary culture with TCA and were quickly applied on 2-DE ; high degree of variety was observed in the 2-DE patterns of exoproteins in S.aureus and GAS ; spots of more than 100 exoproteins of S.aureus and 20 of GAS were identified by the determination of N-terminal amino acid sequences ; a specific spot pattern was identified for each coagulase type of MRSA and variation of the spots of super antigens were observed among the strains belonging to the same coagulase type ; a correlation was observed between the spot patterns and M antigen types in GAS strains ; the MRSA strains that caused NTED and SSSS showed unique toxin production patterns, respectively, observed by 2-DE ; we have accumulated 2-DE patterns of more number of strains that would be helpful for the construction of a 2-DE database of exoproteins.

区分每种细菌菌株，如金黄色葡萄球菌和A组链球菌（GAS），有助于追踪致病菌株和预防医院感染。虽然基于DNA的分型已被普遍用作流行病学方法的歧视，我们开发了二维电泳（2-DE）分析的细菌分泌的外蛋白作为一种新的流行病学方法，我们发现，这种方法是非常适用于区分金黄色葡萄球菌和GAS菌株。确定了培养、生长和提取外蛋白的条件，具有可重复的结果。本研究的主要结果如下：用TCA从早期静止培养物中提取胞外蛋白，并快速应用于2-DE的最佳条件是：金黄色葡萄球菌和GAS胞外蛋白的2-DE图谱具有高度的多样性，通过N端氨基酸序列测定，鉴定出金黄色葡萄球菌100多个胞外蛋白和GAS 20个胞外蛋白的2-DE图谱; 2-DE结果显示，引起NTED和SSSS的MRSA菌株具有独特的产毒模式，而引起NTED和SSSS的MRSA菌株具有独特的产毒模式;本研究积累了更多菌株的双向电泳图谱，为构建胞外蛋白双向电泳数据库奠定了基础。