Study of molecular genetic properties of vaccine strains of anthrax
Journal Title: Бюлетень "Ветеринарна біотехнологія" - Year 2017, Vol 31, Issue
Abstract
Introduction. Molecular-genetic methods widely used to study the genetic peculiarities of microorganisms, one of which is the so-called "RAPD-fingerprint" method, based on using universal short primers that amplify repeating regions of the genome of a microorganism, creating the specific profile of PCR products.The goal of the work. To evaluate the genetic variability of vaccine strains of Bacillus anthracis and saprophytic microorganisms. To choose specific primers for selective amplification of hypervariable fragments of the genome of the anthrax and to optimize the PCR conditions for the differentiation of B. anthracis strains and saprophytic microorganisms of the Bacillus genus.Materials and methods. The vaccine strains B. anthracis CH 05, UA-07, UA-M, and selection of spore-forming saprophytic bacteria of genus Bacillus were used: B. anthrocoides 96,cereus ATCC 10702, B. subtilis 7241. The genotyping of chromosomal DNA was performed using four Specific oligonucleotide primers with a length of 20-22 nucleotides (RAPD-fingerprint).Results of research and discussion. The regimes of PCR were optimized. The used primers amplified a set of major and minor fragments of 3-5 major strips with the size from 800n.b.to 2840 n.b. The set of PCR fragments was the same for all investigated strains of the B.anthracis and consist of 1.0; 1.5; 1.9; 2.3; 2.8 thousand n.b. fragments. The spore-forming saprophytes differed significantly in amplified fragments.Thus, universal primers were complementary to variable repeat domains and was suitable for differentiation of vaccine strains of the anthrax pathogen from saprophytic microorganisms and it could help to detect the possible contamination during preparation of cultures in the production of the vaccine and can be used for differentiation, control and certification of B. anthracis vaccine strains.Conclusions and perspectives of further research. As a result of the studies carried out using specific primers, the strain-specific patterns of amplificates were obtained. Each tested strain was characterized by the presence of both general and individual amplified DNA fragments, differing in molecular weight and specificity. The fragments of the DNA of the vaccine strains of the anthrax pathogen were specifically amplified with primers complementary to the hypervariable sequences of the insertion element of the chromosomal DNA. The method can be used to control the variability of strains during the vaccine preparation
Authors and Affiliations
S. Krylenko, Maria Babkina, Svitlana Tereschenko, Iryna Zotsenko, Basil Sapeiko, Oleksandr Tarasov, Natalia Hudz, Ihor Halka
Keywords
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- EP ID EP266470
- DOI 10.31073/vet_biotech31-10
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