Dorneles Elaine M S, Santana Jordana A, Ribeiro Dayana, Dorella Fernanda Alves, Guimarães Alessandro S, Moawad Mohamed S, Selim Salah A, Garaldi Ana Luiza M, Miyoshi Anderson, Ribeiro Márcio G, Gouveia Aurora M G, Azevedo Vasco, Heinemann Marcos B, Lage Andrey P
Departamento de Medicina Veterinária Preventiva, Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
PLoS One. 2014 Jun 5;9(6):e98758. doi: 10.1371/journal.pone.0098758. eCollection 2014.
The aim of this study was to evaluate the Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR) as a tool for molecular typing of C. pseudotuberculosis isolates from eight different hosts in twelve countries. Ninety-nine C. pseudotuberculosis field strains, one type strain (ATCC 19410T) and one vaccine strain (1002) were fingerprinted using the ERIC-1R and ERIC-2 primers, and the ERIC-1R+ERIC-2 primer pair. Twenty-nine different genotypes were generated by ERIC 1-PCR, 28 by ERIC 2-PCR and 35 by ERIC 1+2-PCR. The discriminatory index calculated for ERIC 1, ERIC 2, and ERIC 1+2-PCR was 0.89, 0.86, and 0.92, respectively. Epidemiological concordance was established for all ERIC-PCR assays. ERIC 1+2-PCR was defined as the best method based on suitability of the amplification patterns and discriminatory index. Minimal spanning tree for ERIC 1+2-PCR revealed three major clonal complexes and clustering around nitrate-positive (biovar Equi) and nitrate-negative (biovar Ovis) strains. Therefore, ERIC 1+2-PCR proved to be the best technique evaluated in this study for genotyping C. pseudotuberculosis strains, due to its usefulness for molecular epidemiology investigations.
本研究的目的是评估肠杆菌重复基因间共有序列（ERIC-PCR）作为一种对来自12个国家8种不同宿主的伪结核棒状杆菌分离株进行分子分型的工具。使用ERIC-1R和ERIC-2引物以及ERIC-1R+ERIC-2引物对，对99株伪结核棒状杆菌野外菌株、1株模式菌株（ATCC 19410T）和1株疫苗菌株（1002）进行指纹图谱分析。ERIC 1-PCR产生了29种不同的基因型，ERIC 2-PCR产生了28种，ERIC 1+2-PCR产生了35种。ERIC 1、ERIC 2和ERIC 1+2-PCR计算出的鉴别指数分别为0.89、0.86和0.92。所有ERIC-PCR检测均建立了流行病学一致性。基于扩增模式的适用性和鉴别指数，ERIC 1+2-PCR被定义为最佳方法。ERIC 1+2-PCR的最小生成树揭示了三个主要的克隆复合体以及围绕硝酸盐阳性（马鼻疽生物变种）和硝酸盐阴性（绵羊生物变种）菌株的聚类。因此，ERIC 1+2-PCR被证明是本研究中评估的用于伪结核棒状杆菌菌株基因分型的最佳技术，因为它对分子流行病学调查有用。
