Comparison of direct PCR and PCR using DNA extracted kit for determine tst and mecA genes in Staphylococcus aureus
Abstract:
Introduction
PCR reactions one of the most accurate and most sensitive tests for the molecular assays. DNA extraction always is time-consuming and costly before the PCR process, which in some cases removed can be saving time and Price. The aim of this study was to compare direct PCR and PCR with DNA extraction template for detection of tst, mecA and femA gens.
Materials And Methods
In this study, clinical isolates of Staphylococcus aureus were confirmed by biochemical tests. Then, with direct PCR by colony acquired of Muller Hinton agar culture and PCR with the extracted DNA extraction kit were studied for tst and mecA genes. For this test, three specific primers were used to target genes.
Findings: amplification was observed of tst, mecA and femA gens with length 326bp, 163bp and 132bp by direct PCR and PCR with the extracted DNA relevant to toxic shock syndrome toxin 1 and are resistant to methicillin. Finally, detected resistant strains and toxins strains by genes. Our results quantitatively Similar each other and in therm of qualitatively were different. The results of PCR test with using extraction kit obtained the better quality.
Conclusions
According to the results obtained in this study, to save time and cost in the detection of Staphylococcus aureus can be used to direct PCR. Bands quality obtained in this way to PCR, DNA, is acceptable and in some cases it can be used as a rapid diagnostic test. To make the better quality must be used mineral oils.
Language:
Persian
Published:
Pars Journal of Medical Sciences, Volume:14 Issue: 3, 2016
Pages:
43 to 46
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