MUTASI T539C GEN pncA (Val180Ala) ISOLAT KLINIS L20 MDR Mycobacterium tuberculosis

Abstract

Abstract: Pyrazinamide is one of the first line drug for therapy of tuberculosis (TB). Isolates L20 of MDR-M. tuberculosis that resistant to pyrazinamide investigated in this research. This research consists of five experimental steps: amplification, electrophoresis, sequencing, and in silico analysis of sequencing results. The results of PCR show a single band of 0,72 kilobase. The nucleotide sequences of isolates L20 and H37Rv have different length of nucleotide sequences, 674 bp and 640 bp, respectively. Nucleotide sequence homology analysis shows that a mutation T539C was observed in the case of the pncA gene of L20 MDR-M. tuberculosis isolate. Substituting mutation towards T539C resulted in changing of amino acid residue Val180Ala. Both amino acids have nonpolar side chain, so that it is not significantly alter three dimensional structure of pyrazinamidase.

Abstrak: Pirazinamida merupakan salah satu obat lini pertama untuk terapi tuberculosis (TB). Tujuan penelitian ini adalah mengkaji Isolat L20 MDR-M. tuberculosis yang resisten terhadap pirazinamida. Penelitian ini meliputi lima tahapan: amplifikasi, elektroforesis, sekuensing, dan analisis hasil sekuensing secara in silico. PCR menghasilkan pita tunggal berukuran 0,72 kilo basa. Sekuen nukleotida isolate L20 dan H37Rv memiliki panjang nukleotida 674 pb dan 640 pb. Analisis homologi sekuen nukleotida menunjukkan bahwa terdapat mutasi T539C pada gen pncA isolat L20 MDR-M. tuberculosis. Mutasi substitusi T539C menghasilkan perubahan residu asam amino Val180Ala yang  sama-sama memiliki rantai samping yang bersifat non polar sehingga secara signifikan tidak mengubah struktur tiga dimensi pirazinamidase.