Comparison of seven techniques for typing international epidemic strains of Clostfidium difficile:: Restriction endonuclease analysis, pulsed-field gel electrophoresis, PCR-ribotyping, multilocus sequence typing, multilocus variable-number tandem-repeat analysis, amplified fragment length polymorphism, and surface layer protein A gene sequence typing
Comparison of seven techniques for typing international epidemic strains of Clostfidium difficile:: Restriction endonuclease analysis, pulsed-field gel electrophoresis, PCR-ribotyping, multilocus sequence typing, multilocus variable-number tandem-repeat analysis, amplified fragment length polymorphism, and surface layer protein A gene sequence typing
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DOI：
10.1128/jcm.01484-07
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发表时间：
2008-02-01
影响因子：
9.4
通讯作者：
McDonald, L. Clifford
中科院分区：
文献类型：
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作者：
Killgore, George;Thompson, Angela;McDonald, L. Clifford
使用加拿大、荷兰、英国和美国实验室提供的42株分离株，我们比较了7种艰难梭菌分型技术的分析结果：多位点可变数目串联重复序列分析（MLVA）、扩增片段长度多态性（AFLP）、表面层蛋白A基因序列分型（slpAST）、PCR-核糖体分型、限制性内切酶分析（REA）、多位点序列分型（MLST）和脉冲场凝胶电泳（PFGE）。我们评估了每种技术的区分能力和分型能力，以及通过AP-F（由毒素型、二元毒素基因的存在和tcdC基因的缺失定义）通过等位基因谱A（AP-A）对分离株进行分组的技术之间的一致性。我们发现，所有的分离株是分型的所有技术和歧视指数得分测试的技术范围从0.964到0.631，在以下顺序。MLVA、REA、PFGE、slpAST、PCR-核糖体分型、MLST和AFLP。所有技术均能区分当前流行的C.艰难梭菌（BI/027/NAP 1）。所有技术均显示AP-A的多种类型（毒素型0，二元毒素阴性，无tcdC基因缺失）。REA、slpAST、MLST和PCR-核糖体分型均包括排除其他AP的单个组中的AP-B（毒素型III，二元毒素阳性，和WC中的18-bp缺失）。PFGE、AFLP和MLVA分别将两个、一个和两个不同的非AP-B分离株与它们的AP-B分离株分组。所有技术似乎都能够检测爆发菌株，但只有REA和MLVA显示出足够的区分能力，以区分不同爆发的菌株。
Using 42 isolates contributed by laboratories in Canada, The Netherlands, the United Kingdom, and the United States, we compared the results of analyses done with seven Clostridium difficile typing techniques: multilocus variable-number tandem-repeat analysis (MLVA), amplified fragment length polymorphism (AFLP), surface layer protein A gene sequence typing (slpAST), PCR-ribotyping, restriction endonuclease analysis (REA), multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE). We assessed the discriminating ability and typeability of each technique as well as the agreement among techniques in grouping isolates by allele profile A (AP-A) through AP-F, which are defined by toxinotype, the presence of the binary toxin gene, and deletion in the tcdC gene. We found that all isolates were typeable by all techniques and that discrimination index scores for the techniques tested ranged from 0.964 to 0.631 in the following order. MLVA, REA, PFGE, slpAST, PCR-ribotyping, MLST, and AFLP. All the techniques were able to distinguish the current epidemic strain of C. difficile (BI/027/NAP1) from other strains. All of the techniques showed multiple types for AP-A (toxinotype 0, binary toxin negative, and no tcdC gene deletion). REA, slpAST, MLST, and PCR-ribotyping all included AP-B (toxinotype III, binary toxin positive, and an 18-bp deletion in WC) in a single group that excluded other APs. PFGE, AFLP, and MLVA grouped two, one, and two different non-AP-B isolates, respectively, with their AP-B isolates. All techniques appear to be capable of detecting outbreak strains, but only REA and MLVA showed sufficient discrimination to distinguish strains from different outbreaks.