Departamento de Microbiologia, Universidade de São Paulo, São Paulo, SP, Brazil.
Res Vet Sci. 2012 Feb;92(1):18-23. doi: 10.1016/j.rvsc.2010.10.025. Epub 2010 Nov 20.
This study evaluated the polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP) analysis of fliC for typing flagella antigen (H) of Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) strains isolated from different animals. The molecular typing of the H type was efficient in the determination of 93 (85%) strains. Two nonmotile (H-) E. coli strains showed a PCR-RFLP electrophoretic profile that did not match known H type patterns. The fliC nucleotide sequence of strains B2N and 4a revealed a nucleotide substitution at the restriction site and a nucleotide insertion that generated a stop codon, respectively. The results of this study showed that PCR-RFLP analysis of fliC is faster, less laborious and as efficient for the determination of H type E. coli isolated from animals, compared to serotyping and that it is useful in determining H type in nonmotile strains and strains expressing non-reactive H antigens. Moreover, the fliC sequence of strain B2N suggests that we could have found a new flagellin antigen type.
本研究评估了聚合酶链反应-限制性片段长度多态性（PCR-RFLP）分析鞭毛抗原（H）在产志贺毒素大肠杆菌（STEC）和肠致病性大肠杆菌（EPEC）分离株从不同动物。H 型的分子分型在确定 93（85%）株方面非常有效。两株非运动（H-）大肠杆菌株显示出与已知 H 型模式不匹配的 PCR-RFLP 电泳图谱。B2N 和 4a 菌株的 fliC 核苷酸序列分别在限制酶切位点处发生核苷酸取代和插入一个终止密码子。本研究结果表明，与血清分型相比，PCR-RFLP 分析 fliC 更快、更省力，对于确定动物来源的大肠杆菌 H 型也同样有效，并且在确定非运动菌株和表达非反应性 H 抗原的菌株的 H 型方面非常有用。此外，B2N 菌株的 fliC 序列表明，我们可能已经发现了一种新的鞭毛抗原类型。
