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Analysis of Clostridium difficile strains causing multiple nosocomial outbreaks in Japan by genotypic typing methods

通过基因分型方法分析引起日本多起医院暴发的艰难梭菌菌株

基本信息

批准号：
14572197
负责人：
KATO Haru
金额：
$ 2.18万
依托单位：
National Institute of Infectious Diseases
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

The typing system sequencing the variable region of the slpA gene was applied to typing Clostridium difficile strains belonging to one PCR ribotype, type smz, which has been identified as frequently causing outbreaks in Japan. The PCR ribotype smz strains recovered from patients at different hospitals in Japan were examined. Among 10 type smz strains tested, three subtypes, smz-1,-2,and -3,were identified that differed from each other by one nucleotide. slpA sequence typing was also applied to direct typing on DNA extracted from stool specimens. Of 22 stool specimens examined, 17 were PCR positive for slpA ; eight were typed as slpA sequence type smz-1,and nine were type smz-2. C.difficile was cultured from 12 of the 17 stool specimens, and the sequence results of the recovered isolates were compared with those from DNA extracted from stool specimens. In all 12 stool specimens, the sequence results of DNA from recovered C.difficile isolates completely agreed with those of DNA extracted directly from stool specimens. The remaining five stool specimens were culture-negative for C.difficile. Sequence typing has the advantage of being easy to compare typing results among multiple laboratories via the Internet without exchanging reference strains to those of typing systems the results of which depend on banding-pattern analyses. slpA sequence typing appears to be a reproducible and reliable typing system for C.difficile as well as being useful for the typing of C.difficile when stool specimens contain only small numbers of C.difficile or are inappropriate for culturing.

对slpA基因可变区测序的分型系统应用于难辨梭菌（Clostridium difficile）的分型，该菌株属于一种PCR核糖型，smz型，已被确定为经常在日本引起疫情。对日本不同医院患者的PCR核糖型smz株进行了检测。在检测的10株smz型菌株中，鉴定出smz-1、-2和-3 3种亚型，它们之间存在1个核苷酸的差异。slpA序列分型也应用于粪便标本提取DNA的直接分型。22份粪便标本中，17份slpA PCR阳性；8个slpA序列为smz-1型，9个为smz-2型。对17份粪便标本中的12份进行艰难梭菌培养，并与粪便标本中提取的DNA序列进行比较。在所有12份粪便标本中，从回收的艰难梭菌分离物中提取的DNA序列结果与直接从粪便标本中提取的DNA序列结果完全一致。其余5例粪便标本难辨梭菌培养阴性。序列分型的优点是可以方便地通过互联网在多个实验室之间比较分型结果，而不需要与依赖带型分析的分型系统交换参考菌株。slpA序列分型似乎是一种可重复且可靠的艰难梭菌分型系统，当粪便标本中只有少量艰难梭菌或不适合培养时，也可用于艰难梭菌分型。