Polymorphism Analysis of the CoA (Coagulase) Gene in Isolates of Methicillin-Resistant Staphylococcus aureus with AluI Restriction Sites

Anita Dwi Anggraini, Eko Budi Koendhori, Hendro Pramono, Daniel Djoko Wahyono

Abstract


Abstract: Analysis of the polymorphism of a gene is important to obtain early information in identifying genetic markers related to the characteristics to be seen. The RFLP method becomes one of the chosen methods because it can see polymorphism that can be detected by using the different fragments of DNA that have been cut by using certain endonuclease enzyme so that it is possible to describe the polymorphism of a gene. Application of RFLP in this study was devoted to see the coagulase gene polymorphism of methichillin-resistant S. aureus. Based on the description above, researcher was interested in conducting the analysis of CoA (coagulase) gene polymorphism with AluI restriction site of methicillin-resistant S. aureus isolates. The results of this study are aimed to be scientific information about the genetic variation of the coagulase gene of methicillin-resistant S. aureus, and as information in the management of diseases related to antibiotic resistance. This was a descriptive study intended to discover the coagulase gene polymorphism of methichillin-resistant S. aureus. The research was conducted in Microbiology Laboratory of Health Analyst Department, Surabaya, and Human Genetic Laboratory of Institute of Tropical Disease of Airlangga University Surabaya. Samples from this study was a collection of isolates of MRSA in Microbiology Laboratory of Dr. Soetomo Hospital Surabaya selected by using consecutive sampling. Based on the bacteriological examination, all of the samples were S. aureus and the results of amplification showed that PCR product (amplicon) of mecA and CoA gene from specific primers, that were 304 bp and 756 bp, had a positivity of 100%. The results of PCR-RFLP of CoA gene showed that all 25 samples underwent polymorphism which was divided into four RFLP patterns and the biggest one was RFLP I pattern (the one which was not digested by AluI restriction enzyme) with a proportion of 64%.


Keywords


Staphylococcus aureus, coA gene, PCR-RFLP, AluI

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