DESAIN IN SILICO DNA PROBE PENDETEKSI MUTASI DAERAH RESISTENSI QUINOLON Gen gyrA DAN gyrB Mycobacterium tuberculosis
| Main Authors: | Pramiswari, Tasya, Tamara, Jennifer, Febrianti, Ni Made, Chandra Yowani, Sagung |
|---|---|
| Format: | Article info application/pdf eJournal |
| Bahasa: | eng |
| Terbitan: |
Jurusan Biologi Universitas Udayana
, 2019
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| Online Access: |
https://ojs.unud.ac.id/index.php/simbiosis/article/view/53532 https://ojs.unud.ac.id/index.php/simbiosis/article/view/53532/31727 |
Daftar Isi:
- Fluoroquinolone (FQ) is the main drug used in MDR-TB therapy resistance to FQ can cause death and increase the risk of treatment failure in MDR-TB patients. Mutations in gyrA gene and gyrB gene from Mycobacterium tuberculosis are responsible for the occurrence of FQ resistance. The highest mutation of gyrA gene in QRDR was found in codon 94, while mutations in gyrB gene was found in codon 500. M. Tuberculosis which resistant to FQ can be detected using the Real Time Polymerase Chain Reaction (RT-PCR) method with DNA probe. This study will design the nucleotide sequence of the TaqMan type probe using the Clone Manager Suite 9.2 program. The results of the DNA probe design were then analyzed in two stages, which is based on the probe criteria in general and based on the TaqMan probe labeling criteria. The design of the mutant probe DNA using the program produced 1 probe for Asp94Ala specific mutations in the gyrA gene and 33 probes for Asp500Ala specific mutations in the gyrB gene. After being analyzed by the two criteria, it was obtained the A94MA1 probe with the 5 '-TCGATCTACGCCAGCCTGGT-3' sequence and B500MA12 probe with the order of 5 '-TACCACAAGCTCGTGCTGATGGC-3'. The results of these probes meet both criteria and can be used to detect mutations in codon 94 gyrA genes and codons 500 gyrB genes of Mycobacterium tuberculosis.
- Fluorokuinolon (FQ) merupakan obat utama yang digunakan pada terapi MDR-TB sehingga resistensi terhadap FQ dapat menyebabkan kematian dan resiko kegagalan terapi pada pasien MDR-TB. Mutasi pada gen gyrA dan gen gyrB dari Mycobacterium tuberculosis bertanggung jawab terhadap terjadinya resistensi FQ. Mutasi region QRDR pada gen gyrA paling tinggi ditemukan pada kodon 94, sedangkan region QRDR pada gen gyrB ditemukan pada kodon 500. Penyakit M. tuberculosis yang resisten terhadap FQ dapat dideteksi menggunakan metode Real Time Polymerase Chain Reaction (RT-PCR) dengan DNA probe. Penelitian ini akan mendesain urutan nukleotida probe berjenis TaqMan menggunakan program Clone Manager Suite 9.2. Hasil rancangan DNA probe kemudian dianalisis 2 tahap yaitu berdasarkan kriteria probe secara umum dan berdasarkan kriteria pelabelan TaqMan probe. Rancangan DNA probe mutan menggunakan program menghasilkan 1 probe untuk mutasi spesifik Asp94Ala pada gen gyrA dan 33 probe untuk mutasi spesifik Asp500Ala pada gen gyrB. Setelah dianalisis dengan kedua kriteria, didapat probe A94MA1 dengan urutan 5’-TCGATCTACGCCAGCCTGGT-3’ dan probe B500MA12 dengan urutan 5’-TACCACAAGCTCGTGCTGATGGC-3’. Hasil probe tersebut memenuhi kedua kriteria dan dapat digunakan untuk mendeteksi mutasi pada kodon 94 gen gyrA dan kodon 500 gen gyrB Mycobacterium tuberculosis.