Достижения науки и техники АПК

Теоретический и научно-практический журнал

2013_12_20_en

DEVELOPMENT OF PCR-TEST SYSTEM FOR DETECTION AND GENOTYPING OF BACTERIA PASTEURELLACEAE FAMILY

 

A.G. Glotov, A.V. Nefedchenko, T.I. Glotova

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Summary. The aim was to develop a test system based on the polymerase chain reaction. To detect the genetic material of bacteria Pasteurella multocida five serogroups and Mannheimia haemolytica specific oligonucleotide primers, selected on the conserved sequences genomes of bacteria in the database NCBI (http://www.ncbi.nlm.nih.gov/), distinguishing them from closely related species and serogroups, were calculated and synthesized. We used three reference strains Pasteurella multocida № №: 1231, 681 and T80, biomaterial samples from sick animals, as well as the culture of bacteria isolated by us during the course of diagnostic studies. The results of development and study of the diagnostic efficiency of the test system based on the polymerase chain reaction for detection and genotyping of Pasteurella multocida and Mannheimia haemolytica, isolated from diseased animals in the territory of Siberia, were demonstrated. Analytical sensitivity of polymerase chain reaction for Pasteurella multocida all serogroups was 104 CFU / ml, and for Mannheimia haemolytica –103 CFU/ml. Polymerase chain reaction is effective at all stages of bacteriological-ray studies. You can use it to differentiate isolates Pas-teurella multocida on genotypes, as well as to distinguish Pasteurella multocida and Mannheimia haemolytica. The predominant genotype of cattle dairy complexes in Siberia is the genotype A bacterium Pasteurella multocida. It is important to consider the presence of different genotypes of circulating bacteria in developing anti-epizootic measures.

Keywords: cattle, respiratory disease, pasteurellosis, polymerase chain reaction, Pasteurella multocida, Mannheimia haemolytica, genotypes.