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中国台湾东北部某医院出现艰难梭菌 PCR 核糖型 127。

The emergence of Clostridioides difficile PCR ribotype 127 at a hospital in northeastern Taiwan.

机构信息

Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan.

Department of Laboratory Medicine, Chang Gung Memorial Hospital at Kaohsiung, Kaohsiung, Taiwan.

出版信息

J Microbiol Immunol Infect. 2022 Oct;55(5):896-909. doi: 10.1016/j.jmii.2021.12.006. Epub 2022 Jan 4.

Abstract

BACKGROUND

Several studies have highlighted the incidence of Clostridioides difficile infections (CDIs) in Taiwan and certain ribotypes have been related to severe clinical diseases. A study was conducted to investigate the polymerase chain reaction (PCR) ribotypes and genetic relatedness of clinical C. difficile strains collected from January 2009 to December 2015 at a hospital in northeastern Taiwan.

MATERIAL AND METHODS

A modified two-step typing algorithm for C. difficile was used by combining a modified 8-plex and 3'-truncated tcdA screening PCR. In addition, MLVA typing was adopted for investigation of bacterial clonality and transmission.

RESULTS

Among a total of 86 strains, 24 (28%) were nontoxigenic and 62 (72%) had both tcdA and tcdB (A + B+). No tcdA-negative and tcdB-positive (AB) strains were identified. Binary toxin (CDT)-producing (cdtA+/cdtB+) strains were started to be identified in 2013. The 21 (34%) AB clinical strains with binary toxin and tcdC deletion were identified as RT127 strains, which contained both RT078-lineage markers and fluoroquinolone (FQ)-resistant mutations (Thr82Ile in gyrA). Multiple loci variable-number tandem repeat analysis (MLVA) for phylogenetic relatedness of RT127 strains indicated that 20 of 21 strains belonged to a clonal complex that was identical to a clinical strain collected from southern Taiwan in 2011, suggestive of a clonal expansion in Taiwan.

CONCLUSION

A two-step typing method could rapidly confirm species identification and define the toxin gene profile of C. difficile isolates. The clonal expansion of RT127 strains in Taiwan indicates monitoring and surveillance of toxigenic C. difficile isolates from human, animal, and environment are critical to develop One Health prevention strategies.

摘要

背景

多项研究强调了台湾地区艰难梭菌感染(CDI)的发生率,某些核糖型与严重的临床疾病有关。本研究旨在调查 2009 年 1 月至 2015 年 12 月期间在台湾东北部一家医院采集的临床艰难梭菌菌株的聚合酶链反应(PCR)核糖型和遗传相关性。

材料与方法

采用改良两步法对艰难梭菌进行分型,结合改良的 8 重和 3'-截断 tcdA 筛查 PCR。此外,还采用 MLVA 对细菌的克隆性和传播进行研究。

结果

在总共 86 株菌株中,24 株(28%)为非产毒株,62 株(72%)同时携带 tcdA 和 tcdB(A+B+)。未发现 tcdA 阴性和 tcdB 阳性(AB)株。2013 年开始鉴定出二元毒素(CDT)产生(cdtA+/cdtB+)菌株。21 株(34%)AB 临床菌株携带二元毒素和 tcdC 缺失,被鉴定为 RT127 株,其同时含有 RT078 谱系标记和氟喹诺酮(FQ)耐药突变(gyrA 中的 Thr82Ile)。RT127 株的多位点可变数串联重复分析(MLVA)表明,21 株中有 20 株属于与 2011 年从台湾南部采集的临床菌株相同的克隆复合体,提示台湾的 RT127 株存在克隆扩张。

结论

两步分型方法可快速确认物种鉴定,并确定艰难梭菌分离株的毒素基因谱。台湾 RT127 株的克隆扩张表明,对人类、动物和环境中的产毒艰难梭菌分离株进行监测和监测,对于制定“同一健康”预防策略至关重要。

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