Polymorphism Analysis of the 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


Latar belakang: Analisis polimorfisme suatu gen penting dilakukan untuk memperoleh informasi lebih awal dalam identifikasi penanda genetik yang berhubungan dengan sifat yang ingin dilihat. Metode RFLP  menjadi salah satu metode yang dipilih karena dapat melihat polimorfisme urutan DNA yang dapat dideteksi melalui adanya perbedaan fragmen DNA setelah dipotong dengan menggunakan enzim endonuclease  tertentu sehingga mampu menggambarkan polimorfisme dari suatu gen. Tujuan penelitian adalah untuk mengetahui adanya polimorfisme gen coagulase S. aureus resisten methicillin.

Metode: Penelitian yang dilakukan secara deskriptif pada 25 sampel. Isolatbakteri MRSA diidentifikasi menggunakan pemeriksaan bakteriologis dan PCR gen mecA dan coagulase dengan menggunakan primer spesifik. Analisis polimorfisme gen coagulase dengan situs restriksi AluI isolat S. aureus resisten methicillin dilakukan dengan menggunakan metoda PCR-RFLP.

Hasil: Amplifikasi menunjukkan produk PCR (amplicon) gen mecA dan coagulase dengan primer spesifik ke 25 isolate bakteri MRSA mempunyai positivitas sebesar 100%. Hasil dari PCR-RFLP menunjukkan empat pola RFLP dengan situs restriksi AluI pada 25 isolat. Proposi terbesar (64%) pada pola RFLP I (pola yang tidak terdigesti enzim restriksi AluI).

Kesimpulan: Terdapat polimorfisme isolate S. aureus resisten methicillin berdasarkan analisis gen coagulase.

Kata kunci: Staphylococcus aureus,gen coagulase, PCR-RFLP, AluI


Background: 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.The aim of the study is to discover the  gene polymorphism of methicillin-resistant Staphylococcus aureus.

Methods: This was a descriptive study using 25 isolates.  Isolates of MRSA tested bybacteriological examination and PCR of mecA and coagulase gene using specific primers.Polymorphism analysis of the coagulase gene in isolates of methicillin-resistant Staphylococcusaureus with AluI Restriction Sites tested by PCR-RFLP.

Results: The Amplification showed that PCR product (amplicon) of mecA and coagulase gene from specific primers of all 25 isolate samples, had a positivity of 100%.The PCR-RFLP of coagulase gene showed that all 25 samples underwent polymorphism into four RFLP patterns with AluI restriction sites. The largest proportion (64%) was found polymorphism in clinical samples MRSA with RFLP I pattern (un-digested pattern of AluI restriction enzyme).

Conclusion: There is polymorphism in the samples MRSA from the analysis of the coagulase gene.

Keywords: Staphylococcus aureus, coagulase gene, PCR-RFLP, AluI


 


Keywords


Staphylococcus aureus, coA gene, PCR-RFLP, AluI

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