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Spa Gene Polymorphism Based Molecular Typing of Clinical MRSA Strains Isolated from Wasit City/ Iraq
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| Author:
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, JASSIM FATEHI ALI, ZAINAB N. AL-SAADI, YOUNIS SAADY SAEED
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| Abstract:
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The investigation goal was to distinguish various methicillin resistant genotypes of Staphylococcus aureus (MRSA)
via using spa gene PCR-RFLP the in Wasit Government. The study started from November 2018 to April 2019,
during which 809 samples were collected from the patients (nose, ear, tonsils, pus, urine, wounds) at Al-Kut
Hospital for Women and Children, Teaching Hospitals of Al-Zahra and Al-Karama and various clinics of medical
at Wasit, they were then processed, cultured. The isolates were examined by cefoxitin (fox) 30 µg disk method
of diffusion and oxacillin (oxa) 1 µg disk method of diffusion. Then, the results confirmed via system of Api Staph
for all isolates resisting to both antibiotics fox and oxa antibiotics, and the results have found that all these
isolates are S. aureus bacteria. The monoplex PCR has been utilized to identify gene of mecA, and to magnify
gene of spa, then the PCR productions were tested via PCR-RFLP using BstH2I as enzyme of restriction for the
first time in Iraq. 78 (37.5%) isolates were not sensitive to both fox and oxa antibiotics, all tested S. aureus isolates
afforded PCR positive for gene of mecA, PCR-RFLP of the gene of spa handed 6 different types of groups
depending on the number of bands and 40 subtypes according to bands MW. After performing a RFLP-PCR
reaction using BstH2I enzyme 40 patterns were obtained from patients according to bands MW. The results also
showed that the value of the discriminatory index for the genotyping of the spa gene was 0.782 for patients
group for all subtypes and patterns. In this study the use of this enzyme (BstH2I) was examined for the first time
in Iraq, where the results were reliable, as this enzyme proved an effective role in the cutting process. Typing of
PCR-RFLP technique is a perfect workable appliance in epidemiological routine observation. It's effortless in
interpretation and achievement, and it can substitute PFGE that is costly and consuming the time.
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| Keyword:
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MRSA, Wasit, Spa gene, Molecular Typing, PCR-RFLP
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| EOI:
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-
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| DOI:
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https://doi.org/10.31838/ijpr/2020.SP3.074
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| Download:
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