An efficient method for identifying Escherichia coli strains isolated from various (Gallus gallus domesticus) organs

详细信息    查看全文

• 作者：V. P. Terletskiy ; V. I. Tyshchenko…
• 刊名：Russian Journal of Genetics: Applied Research
• 出版年：2016
• 出版时间：May 2016
• 年：2016
• 卷：6
• 期：3
• 页码：314-320
• 全文大小：593 KB
• 刊物主题：Human Genetics;
• 出版者：Springer US
• ISSN：2079-0600
• 卷排序：6

文摘

Tracing transmission routes and identifying pathogen sources are important issues in preventive measures aimed at controlling human and animal infectious diseases. A fast and accurate method for bacterial strain identification (genotyping) allows a scientifically sound planning of preventive schemes. Despite the existence of numerous bacteri a genotyping techniques, there is still room for developing a unified typing approach that would be applicable to a variety of bacterial species. The aim is to develop a genotyping method allowing identification of E. coli strains circulating at Russian chicken farms. The method is based on the earlier proposed idea of double digestion and selective labeling of DNA restriction fragments (DDSL). Bacterial genomic DNA is simultaneously digested with two restriction enzymes and labeled with biotinylated deoxynucleoside triphosphates with the presence of DNA polymerase. The enzymes are chosen in silico for each bacterial species so that a limited number of DNA fragments are generated for subsequent separation in conventional agarose gel. After implementation of the study with E. coli isolates, adequate reproducibility and a high discriminatory power of the technique were demonstrated. This approach was previously applied to genotyping other pathogenic bacterial species. The advantages of the proposed technique are the short turnaround time of analysis and easy availability of reagents and equipment. Transmission of a pathogen among chicken within one farm and existence of slightly different E. coli genotypes in various organs of the same individual were observed. Bacterial isolates obtained from any organ except the intestine were suitable for genotyping. Chicken intestine contains endogenous E. coli strains, which hamper the interpretation of the genotyping data obtained for a set of isolates. Thus, our work demonstrates the potential of the DDSL method for genotyping field E. coli isolates in the context of molecular epizootology.Keywordsgenotypingrestriction enzymesDNA digestionlabelingmolecular epizootologyE. coli