Typing by sequencing the slpA gene of Clostridium difficile strains causing multiple outbreaks in Japan
Typing by sequencing the slpA gene of Clostridium difficile strains causing multiple outbreaks in Japan
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DOI：
10.1099/jmm.0.45807-0
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发表时间：
2005-02-01
影响因子：
3
通讯作者：
Arakawa, Y
中科院分区：
文献类型：
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作者：
Kato, H;Yokoyama, T;Arakawa, Y
以前的报道已经证明，表面层蛋白（SIPA）在艰难梭菌分离株之间存在差异。应用sIpA基因可变区测序分型系统对C型进行分型。艰难梭菌菌株属于一种PCR核糖体型，SMZ型，其已被鉴定为在日本经常引起暴发。对从日本不同医院的患者中回收的PCR核糖体型smz菌株进行了检查。在10株SMZ型菌株中，鉴定出3个亚型，即SMZ 1、-2和-3，它们之间仅相差一个核苷酸。slpA序列分型也被应用于从粪便标本中提取的DNA的直接分型。在22份粪便标本中，17份为sIpA PCR阳性; 8份为sIpA序列类型smz-1，9份为smz-2。C.从这17份粪便标本中的12份培养艰难梭菌，并将回收的分离株的序列结果与从粪便标本中提取的DNA的序列结果进行比较。在所有12个粪便标本中，从回收的C.艰难梭菌分离株与直接从粪便标本中提取的DNA完全一致。其余5份粪便标本C培养阴性。很难序列分型的优点是能够通过互联网在多个实验室之间容易地比较分型结果，而不需要像依赖于带型分析的分型系统中所要求的那样交换参考菌株。sIpA序列分型是一种可重复的、可靠的C.艰难梭菌，并可用于分型的C。当粪便标本仅含有少量C.难以培养或不适合培养。
Previous reports have documented that a surface layer protein (SIpA) varies among Clostridium difficile isolates. The typing system by sequencing the variable region of the sIpA gene was applied to typing C. 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 sIpA; eight were typed as sIpA sequence type smz-1 and nine as type smz-2. C. difficile was cultured from 12 of these 17 stool specimens, and the sequence results of the recovered isolates were compared with those from the DNA extracted from the stool specimens. In all 12 of these 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 enabling easy comparison of typing results among multiple laboratories via the Internet without exchanging reference strains as is required in typing systems which depend on banding-pattern analyses. sIpA 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.