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...
Main Authors: | , , , , , |
---|---|
Format: | Article |
Language: | Russian |
Published: |
Federal Government Health Institution, Russian Research Anti-Plague Institute “Microbe”
2018-10-01
|
Series: | Проблемы особо опасных инфекций |
Subjects: | |
Online Access: | https://journal.microbe.ru/jour/article/view/991 |
id |
doaj-2f1389dfefed4d8289b0a9aa23ce47a5 |
---|---|
record_format |
Article |
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 |
work_keys_str_mv |
AT nikhammadov markerlociinbrucellagenomefordifferentialpcrindicationofpathogenicstrains AT kaosyanin markerlociinbrucellagenomefordifferentialpcrindicationofpathogenicstrains AT kvusoltsev markerlociinbrucellagenomefordifferentialpcrindicationofpathogenicstrains AT tkhfaizov markerlociinbrucellagenomefordifferentialpcrindicationofpathogenicstrains AT avkhammadova markerlociinbrucellagenomefordifferentialpcrindicationofpathogenicstrains AT eashuralev markerlociinbrucellagenomefordifferentialpcrindicationofpathogenicstrains |
_version_ |
1721291724811141120 |