Tracing of transmission routes and identification of 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 scientifically sound planning of preventive schemes. Despite the existence of numerous bacterium 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 be generated for subsequent separation in conventional agarose gel. After implementation of the study with E. coli isolates, adequate reproducibility and 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 turn-around 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 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.
About The Authors
V. P. Terletskiy. Russian Research Institute of Farm Animal Genetics and Breeding, Federal Agency for Scientific Organizations, St.-Petersburg, Pushkin, Russia All-Russia Research Veterinary Institute of Poultry Science, Federal Agency for Scientific Organizations, St.-Petersburg, Lomonosov, Russia, Russian Federation
O. B. Novikova. All-Russia Research Veterinary Institute of Poultry Science, Federal Agency for Scientific Organizations, St.-Petersburg, Lomonosov, Russia, Russian Federation
I. Ya. Shakhtamirov. Chechen State University, Ministry of Education and Science of the Russian Federation, Groznyi, Chechen Republic, Russia, Russian Federation
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