Carvalho de Moura A C, Irino K, Vidotto M C
Universidade Estadual de Londrina, Departamento de Microbiologia, Londrina, PR, Brazil.
Avian Dis. 2001 Jan-Mar;45(1):173-81.
In this study, we tested the capability of enterobacterial repetitive intergenic consensus (ERIC) and repetitive extragenic palindromic (REP) polymerase chain reaction (PCR) to detect genetic diversity among Escherichia coli strains isolated from chickens bearing clinical signs of colibacillosis and compared the genotypes so obtained with the O:H serotypes and virulence of those strains. The DNAs from 50 avian E. coli strains and from E. coli ATCC 25922 were used to amplify ERIC and REP sequences. DNA from avian strains produced from 8 to 17 bands by ERIC-PCR and from 6 to 20 bands by REP-PCR; E. coli ATCC produced 11 bands by both methods. ERIC and REP-PCR showed good discriminating power, and the dendograms based on the different patterns revealed extensive genetic diversity among the avian strains. Those strains were allocated into four major clonal clusters, each one with 60% of similarity by ERIC and REP-PCR, and those clusters corresponded to strains with different degrees of pathogenicity. However, 56% of the pathogenic strains (28/50) belonged to two out of three major clonal clusters, and 86% of the nonpathogenic strains tended to group in one cluster and one subgroup. The 32 serotypes detected were distributed in all clusters, and within a serogroup, different DNA fingerprints were observed; however, strains with same serotypes tended to form clusters with similarity coefficients greater than 80%. These results suggest that no specific serotype and genotype is responsible for colibacillosis and that REP and ERIC-PCR are reproducible techniques that can improve the studies needed to clarify the pathways to the pathogenesis of colibacillosis.
在本研究中,我们测试了肠杆菌重复基因间共识序列(ERIC)和重复基因外回文序列(REP)聚合酶链反应(PCR)检测从患有大肠杆菌病临床症状的鸡中分离出的大肠杆菌菌株间遗传多样性的能力,并将由此获得的基因型与这些菌株的O:H血清型和毒力进行比较。使用来自50株禽源大肠杆菌菌株和大肠杆菌ATCC 25922的DNA扩增ERIC和REP序列。禽源菌株的DNA经ERIC-PCR产生8至17条带,经REP-PCR产生6至20条带;大肠杆菌ATCC经两种方法均产生11条带。ERIC和REP-PCR显示出良好的鉴别能力,基于不同模式的树状图揭示了禽源菌株间广泛的遗传多样性。这些菌株被分为四个主要克隆簇,通过ERIC和REP-PCR每个克隆簇的相似度为60%,且这些簇对应于不同致病程度的菌株。然而,56%的致病菌株(28/50)属于三个主要克隆簇中的两个,86%的非致病菌株倾向于聚集在一个簇和一个亚组中。检测到的32种血清型分布在所有簇中,并且在一个血清群内,观察到不同的DNA指纹图谱;然而,相同血清型的菌株倾向于形成相似度系数大于80%的簇。这些结果表明,没有特定的血清型和基因型是导致大肠杆菌病的原因,并且REP和ERIC-PCR是可重复的技术,能够改进阐明大肠杆菌病发病机制途径所需的研究。