Marker Loci in Brucella Genome for Differential PCR Indication of Pathogenic strains

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 amp...

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Main Authors: N. I. Khammadov, K. A. Osyanin, K. V. Usol’tsev, T. Kh. Faizov, A. V. Khammadova, E. A. Shuralev
Format: Article
Language:Russian
Published: Federal Government Health Institution, Russian Research Anti-Plague Institute “Microbe” 2018-10-01
Series:Проблемы особо опасных инфекций
Subjects:
pcr
Online Access:https://journal.microbe.ru/jour/article/view/991
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spelling doaj-2f1389dfefed4d8289b0a9aa23ce47a52021-07-22T07:51:41ZrusFederal Government Health Institution, Russian Research Anti-Plague Institute “Microbe”Проблемы особо опасных инфекций0370-10692658-719X2018-10-0103889310.21055/0370-1069-2018-3-88-93975Marker Loci in Brucella Genome for Differential PCR Indication of Pathogenic strainsN. I. Khammadov0K. A. Osyanin1K. V. Usol’tsev2T. Kh. Faizov3A. V. Khammadova4E. A. Shuralev5Federal Center for Toxicological, Radiation and Biological Safety, KazanFederal Center for Toxicological, Radiation and Biological Safety, KazanFederal Center for Toxicological, Radiation and Biological Safety, KazanFederal Center for Toxicological, Radiation and Biological Safety, KazanKazan Federal University, KazanFederal Center for Toxicological, Radiation and Biological Safety, Kazan; Kazan Federal University, Kazan; Kazan State Medical Academy, KazanObjective 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.https://journal.microbe.ru/jour/article/view/991brucellaindicationdifferentiationspeciesstrainpcrmlva
collection DOAJ
language Russian
format Article
sources DOAJ
author N. I. Khammadov
K. A. Osyanin
K. V. Usol’tsev
T. Kh. Faizov
A. V. Khammadova
E. A. Shuralev
spellingShingle N. I. Khammadov
K. A. Osyanin
K. V. Usol’tsev
T. Kh. Faizov
A. V. Khammadova
E. A. Shuralev
Marker Loci in Brucella Genome for Differential PCR Indication of Pathogenic strains
Проблемы особо опасных инфекций
brucella
indication
differentiation
species
strain
pcr
mlva
author_facet N. I. Khammadov
K. A. Osyanin
K. V. Usol’tsev
T. Kh. Faizov
A. V. Khammadova
E. A. Shuralev
author_sort N. I. Khammadov
title Marker Loci in Brucella Genome for Differential PCR Indication of Pathogenic strains
title_short Marker Loci in Brucella Genome for Differential PCR Indication of Pathogenic strains
title_full Marker Loci in Brucella Genome for Differential PCR Indication of Pathogenic strains
title_fullStr Marker Loci in Brucella Genome for Differential PCR Indication of Pathogenic strains
title_full_unstemmed Marker Loci in Brucella Genome for Differential PCR Indication of Pathogenic strains
title_sort marker loci in brucella genome for differential pcr indication of pathogenic strains
publisher Federal Government Health Institution, Russian Research Anti-Plague Institute “Microbe”
series Проблемы особо опасных инфекций
issn 0370-1069
2658-719X
publishDate 2018-10-01
description 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.
topic brucella
indication
differentiation
species
strain
pcr
mlva
url https://journal.microbe.ru/jour/article/view/991
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