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Ni Made Febrianti
Program Studi Farmasi, Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Udayana
Agriculture, Biological Sciences & Forestry

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IN SILICO DNA PROBE DESIGN FOR MUTATION DETECTION OF QUINOLON RESISTANCE AREA gyrA AND gyrB GENE Mycobacterium tuberculosis Tasya Pramiswari; Jennifer Tamara; Ni Made Febrianti; Sagung Chandra Yowani
SIMBIOSIS Vol 7 No 2 (2019)
Publisher : Jurusan Biologi Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (352.871 KB) | DOI: 10.24843/JSIMBIOSIS.2019.v07.i02.p05

Abstract

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.
Co-Authors Jennifer Tamara Sagun Chandra Yowani Tasya Pramiswari