Izvestiya of Saratov University.

Chemistry. Biology. Ecology

ISSN 1816-9775 (Print)
ISSN 2541-8971 (Online)


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Russian
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Article type: 
Article
UDC: 
616.98:579.842.23

Differentiation of Yersinia pestis Strains of the Main Subspecies of Medieval Biovar by Real-Time PCR Method with Hybridization-Fluorescenсe Registration and SNP Typing

Autors: 
Balykova Alina N., Saratov State University
Nosov Nikita Yu., Russian Research Anti-Plague Institute "Microbe"
Nikiforov Konstantin A., Russian Research Anti-Plague Institute "Microbe"
Eroshenko Galina A., Russian Research Anti-Plague Institute "Microbe"
Abstract: 

Methods of differentiation and identification of Yersinia pestis strains of medieval biovar by real-time PCR method with hybridization-fluorescent results’ registration and SNP typing method have been developed. Ninety-one Y. pestis strains of medieval biovar from the plague foci of Russia and other CIS countries, received from the State Collection of Pathogenic Bacteria at the Russian Research Anti-Plague Institute “Microbe”, were used in this work. The strains of medieval biovar are represented by four already known phylogenetic lines: 2.MED0, 2.MED1, 2.MED2, 2.MED3. As a result of phylogenetic analysis we have discovered a new phylogenetic line, named 2.MED4. We have found 3 SNPs, which are marker for this line and located in protein-coding regions of the genome, which can be used to establish the belonging of Y. pestis strains of the medieval biovar to the line 2.MED4. To identify and differentiate strains of the medieval biovar by the multiplex realtime PCR method with hybridization-fluorescent results’registration, 4 DNA-targets: Med24, pCKF, 2.MED1, 2.MED3 have been selected. 100% specificity of these targets is shown. The use of a complex of the developed methods of real-time PCR and SNP typing provides a successful differentiation of strains of the medieval biovar by phylogeographic affiliation.

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