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脉冲场凝胶电泳、肠杆菌重复基因间共有序列PCR及生化试验用于鉴定格氏乳球菌的比较

Comparison of pulsed-field gel electrophoresis and enterobacterial repetitive intergenic consensus PCR and biochemical tests to characterize Lactococcus garvieae.

作者信息

Ture M, Altinok I, Capkin E

出版信息

J Fish Dis. 2015 Jan;38(1):37-47. doi: 10.1111/jfd.12192.

Abstract

Biochemical test, pulsed-field gel electrophoresis (PFGE) and enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) were used to compare 42 strains of Lactococcus garvieae isolated from different regions of Turkey, Italy, France and Spain. Twenty biotypes of L. garvieae were formed based on 54 biochemical tests. ERIC-PCR of genomic DNA from different L. garvieae strains resulted in amplification of multiple fragments of DNA in sizes ranging between 200 and 5000 bp with various band intensities. After cutting DNA with ApaI restriction enzyme and running on the PFGE, 11–22 resolvable bands ranging from 2 to 194 kb were observed. Turkish isolates were grouped into two clusters, and only A58 (Italy) strain was connected with Turkish isolates. Similarities between Turkish, Spanish, Italian and French isolates were <50% except 216-6 Rize strain. In Turkey, first lactococcosis occurred in Mugla, and then, it has been spread all over the country. Based on ERIC-PCR, Spanish and Italian strains of L. garvieae were related to Mugla strains. Therefore, after comparing PFGE profiles, ERIC-PCR profiles and phenotypic characteristics of 42 strains of L. garvieae, there were no relationships found between these three typing methods. PFGE method was more discriminative than the other methods.

摘要

采用生化试验、脉冲场凝胶电泳(PFGE)和肠杆菌基因间重复一致序列聚合酶链反应(ERIC-PCR)对从土耳其、意大利、法国和西班牙不同地区分离出的42株格氏乳球菌进行比较。基于54项生化试验形成了20种格氏乳球菌生物型。对不同格氏乳球菌菌株的基因组DNA进行ERIC-PCR,结果扩增出大小在200至5000 bp之间的多个DNA片段,条带强度各异。用ApaI限制性内切酶切割DNA并进行PFGE后,观察到11 - 22条可分辨条带,大小在2至194 kb之间。土耳其分离株被分为两个簇,只有A58(意大利)菌株与土耳其分离株相连。除216 - 6里泽菌株外,土耳其、西班牙、意大利和法国分离株之间的相似性<50%。在土耳其,首次乳球菌病发生在穆拉,随后蔓延至全国。基于ERIC-PCR,西班牙和意大利的格氏乳球菌菌株与穆拉菌株相关。因此比较42株格氏乳球菌的PFGE图谱、ERIC-PCR图谱和表型特征后,发现这三种分型方法之间没有关联。PFGE方法比其他方法更具鉴别力。

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