Volume 5, Issue 3 (2019)
IEM 2019, 5(3): 27-38 |
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hoseiny khorram abadi S E, zaker bostan abad S. Molecular Characterization of Staphylococcus aureus Isolated from Clinical Samples Based on 16srRNA, rpoB, and hsp70 Genes by MLSA
. IEM 2019; 5 (3) :27-38
URL: http://iem.modares.ac.ir/article-4-35116-en.html
URL: http://iem.modares.ac.ir/article-4-35116-en.html
1- Department of biology, school of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran
2- Department of biotechnology, School of Basic Sciences, Parand Branch, Islamic Azad University, Tehran, Iran , saeedzaker20@yahoo.com
2- Department of biotechnology, School of Basic Sciences, Parand Branch, Islamic Azad University, Tehran, Iran , saeedzaker20@yahoo.com
Abstract: (3358 Views)
Aims: Staphylococcus aureus is a Gram-positive bacterium with the capability of causing a variety of nosocomial and community-acquired infections. Evaluating the genetic structure, polymorphism, genotyping, and phylogeny of S. aureus isolates could contribute to the prevention and treatment of infections caused by this microorganism.
Materials & Methods: In this study, the polymorphisms of 16S rRNA, rpoB, and hsp70 genes were investigated in a total of 50 S. aureus isolates using S. aureus NCTC 8325 as the reference strain. Polymerase chain reaction (PCR) was used for the detection and amplification of the studied genes. The amplicons were then sequenced using a Sanger sequencing method. Moreover, phylogeny of the isolates was studied using Neighbor-joining and Maximum Parsimony methods for 16S rRNA, rpoB, and hsp70 genes individually and in combination.
Findings: After Sanger sequencing, data obtained by Sequencher and Mesquite software programs revealed several polymorphisms of S. aureus isolates 16S rRNA, rpoB, and hsp70 genes, respectively. These polymorphisms included transversion, transition, insertion, and deletion. Among the studied strains, 10 cases showed no polymorphism. Multi-locus sequence analysis (MLSA) showed several genetic diversities in S. aureus isolates.
Conclusion: It seems essential to rapidly and reliably identify the phylogenetic sources and characteristics of this microorganism and to have a better understanding of its molecular epidemiology in order for infection practical surveillance and control.
Materials & Methods: In this study, the polymorphisms of 16S rRNA, rpoB, and hsp70 genes were investigated in a total of 50 S. aureus isolates using S. aureus NCTC 8325 as the reference strain. Polymerase chain reaction (PCR) was used for the detection and amplification of the studied genes. The amplicons were then sequenced using a Sanger sequencing method. Moreover, phylogeny of the isolates was studied using Neighbor-joining and Maximum Parsimony methods for 16S rRNA, rpoB, and hsp70 genes individually and in combination.
Findings: After Sanger sequencing, data obtained by Sequencher and Mesquite software programs revealed several polymorphisms of S. aureus isolates 16S rRNA, rpoB, and hsp70 genes, respectively. These polymorphisms included transversion, transition, insertion, and deletion. Among the studied strains, 10 cases showed no polymorphism. Multi-locus sequence analysis (MLSA) showed several genetic diversities in S. aureus isolates.
Conclusion: It seems essential to rapidly and reliably identify the phylogenetic sources and characteristics of this microorganism and to have a better understanding of its molecular epidemiology in order for infection practical surveillance and control.
Article Type: Original Research |
Subject:
Bacteriology
Received: 2019/07/24 | Accepted: 2019/09/26 | Published: 2020/01/8
Received: 2019/07/24 | Accepted: 2019/09/26 | Published: 2020/01/8
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