Department of Fishery Microbiology, Karnataka Veterinary, Animal & Fisheries Sciences University, College of Fisheries, Mangalore, India.
Indian J Med Res. 2012 Mar;135(3):371-81.
BACKGROUND & OBJECTIVES: Infections due to seafood associated Salmonella serovars are great risk to public health. Different phenotypic characteristics have been used previously for epidemiological investigation of Salmonella. Beyond the phenotypic characterization, a reliable genetic level discriminatory method is required. Therefore, this study was attempted to use different phenotypic and molecular fingerprinting methods for investigation of genetic diversity among seafood associated nontyphoidal Salmonella serovars.
Fifty eight seafood associated Salmonella isolates were included in this study. All isolates were serotyped and epidemiological investigation was carried out using molecular fingerprinting methods, random amplified polymorphic DNA (RAPD) and enterobacterial repetitive intergenic consensus sequence based-PCR (ERIC-PCR) along with whole cell protein profiling using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) in our study.
Among the 58 Salmonella isolates, S. Weltevreden was observed to be the most predominant serovar. Typing of Salmonella serovars using RAPD and ERIC-PCR suggested the existence of a genetic diversity. Though both PCR based techniques were found to have a good discriminatory index, a better discriminatory ability was observed when the results obtained by the two techniques were combined and taken for composite analysis. Protein profiling of whole cells using SDS-PAGE demonstrated the presence of several bands with two bands of sizes 38 kDa and 46 kDa common among all 58 isolates.
INTERPRETATION & CONCLUSIONS: Our study shows that use of protein profiling in combination with established typing methods such as RAPD and ERIC-PCR may provide useful information in typing of non-typhoidal Salmonella isolates associated with seafood and to develop strategies to protect public from Salmonella infections.
与海产品相关的沙门氏菌血清型引起的感染对公众健康构成了巨大威胁。先前已经使用不同的表型特征来进行沙门氏菌的流行病学研究。除了表型特征外，还需要一种可靠的遗传水平区分方法。因此，本研究尝试使用不同的表型和分子指纹分析方法来调查与海产品相关的非伤寒沙门氏菌血清型的遗传多样性。
本研究纳入了 58 株与海产品相关的沙门氏菌分离株。所有分离株均进行血清分型，并通过分子指纹分析方法（随机扩增多态性 DNA 分析（RAPD）和肠杆菌重复基因间 consensus 序列基 PCR（ERIC-PCR））以及使用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）进行全细胞蛋白谱分析进行流行病学调查。
在 58 株沙门氏菌分离株中，观察到 Weltevreden 血清型最为常见。使用 RAPD 和 ERIC-PCR 对沙门氏菌血清型进行分型表明存在遗传多样性。尽管两种基于 PCR 的技术都具有良好的区分指数，但当将两种技术的结果结合并进行综合分析时，观察到更好的区分能力。使用 SDS-PAGE 对全细胞蛋白进行谱分析显示存在多个条带，其中 38 kDa 和 46 kDa 两个大小的条带在所有 58 个分离株中均存在。
本研究表明，使用蛋白谱分析结合 RAPD 和 ERIC-PCR 等已建立的分型方法可能为与海产品相关的非伤寒沙门氏菌分离株的分型提供有用信息，并制定策略以保护公众免受沙门氏菌感染。
