Marker Loci in Brucella Genome for Differential PCR Indication of Pathogenic strains
https://doi.org/10.21055/0370-1069-2018-3-88-93
Abstract
Objective of this work was to develop the algorithms for differential PCR indication of Brucella genus strains using databases of their genomes. Materials and methods .Resources of the National Center for Biotechnology Information (NCBI) and BLAST and Vector NTI 9.1.0 software utilities. For PCR amplification, B. suis, B. abortus, B. melitensis nucleic acids, as well as plasmid DNA with marker insertions were used. Results and conclusions. We assessed brucella gene sequences, some of which are found in Brucella genus bacteria, others only in representatives of B. melitensis, and the third ones – only in representatives of B. abortus. As a result of primers and probes designing for indication of Brucella genus bacteria and representatives of B. melitensis and B. abortus species, criteria for marker sequence amplification have been established. These criteria provide for simultaneous differentiation in a single reaction. The determination of strain differences within one species of Brucella is described in multilocus VNTR assay technique, and the profiles of tandem repeats of various B. melitensis and B. abortus strains are available in the public domain. To monitor the progress of amplification, a positive control has been developed that has the nucleotide sequence of all marker regions. The text of the paper discloses all the nucleotide sequences of primers, probes and positive control, which makes it possible to independently acquire them in competent organizations.
About the Authors
N. I. Khammadov
Federal Center for Toxicological, Radiation and Biological Safety, Kazan
Russian Federation
Russian Federation
Nauchniy Gorodok-2, Kazan, Tatarstan, 420075
K. A. Osyanin
Federal Center for Toxicological, Radiation and Biological Safety, Kazan
Russian Federation
Russian Federation
Nauchniy Gorodok-2, Kazan, Tatarstan, 420075
K. V. Usol’tsev
Federal Center for Toxicological, Radiation and Biological Safety, Kazan
Russian Federation
Nauchniy Gorodok-2, Kazan, Tatarstan, 420075
Russian Federation
Nauchniy Gorodok-2, Kazan, Tatarstan, 420075
T. Kh. Faizov
Federal Center for Toxicological, Radiation and Biological Safety, Kazan
Russian Federation
Nauchniy Gorodok-2, Kazan, Tatarstan, 420075
Russian Federation
Nauchniy Gorodok-2, Kazan, Tatarstan, 420075
A. V. Khammadova
Kazan Federal University, Kazan
Russian Federation
Russian Federation
18, Kremlyovskaya St., Kazan, Tatarstan, 420008
E. A. Shuralev
Federal Center for Toxicological, Radiation and Biological Safety, Kazan; Kazan Federal University, Kazan; Kazan State Medical Academy, Kazan
Russian Federation
Russian Federation
Federal Center for Toxicological, Radiation and Biological Safety; Nauchniy Gorodok-2, Kazan, Tatarstan, 420075, Russian Federation.
Kazan Federal University; 18, Kremlyovskaya St., Kazan, Tatarstan, 420008, Russian Federation.
Kazan State Medical Academy; 36, Butlerova St., Kazan, Tatarstan, 420012, Russian Federation
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Review
For citations:
Khammadov N.I., Osyanin K.A., Usol’tsev K.V., Faizov T.Kh., Khammadova A.V., Shuralev E.A. Marker Loci in Brucella Genome for Differential PCR Indication of Pathogenic strains. Problems of Particularly Dangerous Infections. 2018;(3):88-93. (In Russ.) https://doi.org/10.21055/0370-1069-2018-3-88-93
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