Cepheid, Sunnyvale, California 94089, USA.
J Clin Microbiol. 2011 May;49(5):1831-7. doi: 10.1128/JCM.02446-10. Epub 2011 Mar 9.
Accurate strain typing is critical for understanding the changing epidemiology of Clostridium difficile infections. We typed 350 isolates of toxigenic C. difficile from 2008 to 2009 from seven laboratories in the United States and Canada. Typing was performed by PCR-ribotyping, pulsed-field gel electrophoresis (PFGE), and restriction endonuclease analysis (REA) of whole-cell DNA. The Cepheid Xpert C. difficile test for presumptive identification of 027/NAP1/BI isolates was also tested directly on original stool samples. Of 350 isolates, 244 (70%) were known PCR ribotypes, 224 (68%) were 1 of 8 common REA groups, and 187 (54%) were known PFGE types. Eighty-four isolates typed as 027, NAP1, and BI, and 83 of these were identified as presumptive 027/NAP1/BI by Xpert C. difficile. Eight additional isolates were called presumptive 027/NAP1/BI by Xpert C. difficile, of which three were ribotype 027. Five PCR ribotypes contained multiple REA groups, and three North American pulsed-field (NAP) profiles contained both multiple REA groups and PCR ribotypes. There was modest concordance of results among the three methods for C. difficile strains, including the J strain (ribotype 001 and PFGE NAP2), the toxin A-negative 017 strain (PFGE NAP9 and REA type CF), the 078 animal strain (PFGE NAP7 and REA type BK), and type 106 (PFGE NAP11 and REA type DH). PCR-ribotyping, REA, and PFGE provide different but overlapping patterns of strain clustering. Unlike the other methods, the Xpert C. difficile 027/NAP1/BI assay gave results directly from stool specimens, required only 45 min to complete, but was limited to detection of a single strain type.
准确的菌株分型对于了解艰难梭菌感染的不断变化的流行病学至关重要。我们对来自美国和加拿大的 7 个实验室的 2008 年至 2009 年间的 350 株产毒艰难梭菌进行了分型。通过 PCR-核糖体分型、脉冲场凝胶电泳 (PFGE) 和全细胞 DNA 的限制性内切酶分析 (REA) 进行分型。还直接在原始粪便样本上测试了 Cepheid Xpert C. difficile 检测对 027/NAP1/BI 分离株的推定鉴定。在 350 株分离株中,244 株(70%)为已知的 PCR 核糖体型,224 株(68%)为 8 个常见 REA 组之一,187 株(54%)为已知的 PFGE 型。84 株分离株为 027、NAP1 和 BI 型,其中 83 株通过 Xpert C. difficile 鉴定为推定的 027/NAP1/BI。另外 8 株通过 Xpert C. difficile 鉴定为推定的 027/NAP1/BI,其中 3 株为 027 核糖体型。5 种 PCR 核糖体型包含多个 REA 组,3 种北美脉冲场 (NAP) 图谱包含多个 REA 组和 PCR 核糖体型。三种方法对艰难梭菌菌株的结果存在适度的一致性,包括 J 株(核糖体型 001 和 PFGE NAP2)、毒素 A 阴性 017 株(PFGE NAP9 和 REA 型 CF)、078 动物株(PFGE NAP7 和 REA 型 BK)和 106 型(PFGE NAP11 和 REA 型 DH)。PCR-核糖体分型、REA 和 PFGE 提供了不同但重叠的菌株聚类模式。与其他方法不同,Xpert C. difficile 027/NAP1/BI 检测直接从粪便标本中获得结果,仅需 45 分钟即可完成,但仅限于检测单一菌株类型。