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Comparison of PCR-RFLP and PFGE for determining the clonality of Brucella isolates from human and livestock specimens
- Source :
- Saudi Journal of Biological Sciences, Vol 26, Iss 2, Pp 256-262 (2019)
- Publication Year :
- 2019
- Publisher :
- Elsevier, 2019.
-
Abstract
- Brucellosis is an important zoonotic disease caused by different species of genus Brucella that are pathogenic for humans and a variety of animals. Accurate detection of Brucella spp. infection is important for control of disease. The aim of this study was to comparison of molecular genotyping of Brucella strains by Pulsed-field gel electrophoresis (PFGE) and polymerase chain reaction -Restriction Fragment Length Polymorphism (PCR-RFLP) techniques.Twenty- seven Brucella spp. were isolated from human and animal samples. The isolates identified by conventional microbiological methods and confirmed using PCR for amplification of omp2a gene. Molecular typing of Brucella strains carried out by PCR-RFLP after PstI and PFGE of chromosomal DNA after XbaI enzyme digestion. The omp2a gene PCR Products with different patterns of PCR-RFLP were sequenced.The omp2a gene amplification of all human and animal Brucella isolates were positive for 1100 bp fragment. By PCR-RFLP analysis two genotypes/patterns for human isolates and four genotypes for animal isolates were obtained. In PFGE analysis totally, 7 common clones/clusters and 3 single clones were obtained.The results of this study showed the PFGE method is the more reliable and useful assay for molecular typing of Brucella strains and is more preferred to PCR-RFLP in determination of genetic similarity among human and animal Brucella isolates. The presented data showed PCR-RFLP analysis was not able to differentiate between B. melitensis biovars and vaccine strain. Keywords: Brucella melitensis, PCR, RFLP, Brucellosis, Molecular typing
- Subjects :
- 0301 basic medicine
Gel electrophoresis
biology
030106 microbiology
Brucella
biology.organism_classification
bacterial infections and mycoses
Virology
law.invention
Microbiology
03 medical and health sciences
lcsh:Biology (General)
law
Genotype
Gene duplication
Pulsed-field gel electrophoresis
Restriction fragment length polymorphism
General Agricultural and Biological Sciences
lcsh:QH301-705.5
Polymerase chain reaction
Brucella melitensis
Subjects
Details
- Language :
- English
- Volume :
- 26
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Saudi Journal of Biological Sciences
- Accession number :
- edsair.doi.dedup.....5a46bc73aa121c34453f5f2238d567bb