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.

Highlights

  • Fluoroquinolone (FQ) is the main drug used in Multidrug Resistant Tuberculosis (MDR-TB) therapy resistance to FQ can cause death and increase the risk of treatment failure in MDR-TB patients

  • the main drug used in MDR-TB therapy resistance to FQ can cause death

  • Mutations in gyrA gene and gyrB gene from Mycobacterium tuberculosis are responsible for the occurrence of FQ resistance

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Summary

Program Studi Biologi FMIPA UNUD

ABSTRAK 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. Penyakit M. tuberculosis yang resisten terhadap FQ dapat dideteksi menggunakan metode Real Time Polymerase Chain Reaction (RT-PCR) dengan DNA probe. 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. Hasil probe tersebut memenuhi kedua kriteria dan dapat digunakan untuk mendeteksi mutasi pada kodon 94 gen gyrA dan kodon 500 gen gyrB Mycobacterium tuberculosis. Kata kunci: MDR-TB, gen gyrA, gen gyrB, TaqMan probe, Real-Time PCR

MATERI DAN METODE
Penentuan urutan nukleotida target
Analisis primer
Analisis hasil perancangan DNA probe
Tidak ada
Oligonucleotides with Fluorescent
Tuberculosis pada Isolat Klinik
Full Text
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