Deteksi Mycobacterium tuberculosis dan resistensinya terhadap rifampisin dengan Metode nested polymerase chain reaction (PCR) dan sequencing

Main Article Content

Maria Lina Rosilawati

Abstract

Latar Belakang
DNA rpob (RNA polymerase sub unit b) Mycobacterium tuberculosis dapat diamplifikasi secara spesifik dengan metode nested polymerase chain reaction (PCR). Nested PCR yang dilanjutkan dengan sequencing dapat secara langsung diaplikasikan untuk mendeteksi M. tuberculosis dan menentukan mutasi pada gen rpob yang berkaitan dengan resistensinya terhadap rifampisin, pada isolat klinis maupun sampel sputum.

Metode
Dalam penelitian ini digunakan 20 isolat klinis dan 30 sampel sputum yang diamplifikasi dengan primer yang dirancang dari bagian gen rpob M. tuberculosis. Metode fenol-kloroform dan metode Boom masing-masing digunakan untuk ekstraksi DNA isolat klinis dan sampel sputum. Sequencing hanya dilakukan untuk hasil PCR dari sampel sputum.

Hasil
Dari 20 isolat klinis, 15 isolat positif terdeteksi sebagai M. tuberculosis dengan nested PCR, 4 isolat tergolong Mycobacteria other than tuberculosis (MOTT) dan 1 isolat non-Mycobacteria. Hasil nested PCR pada 30 sampel sputum dengan 25 sampel basil tahan asam (BTA) positif dan 5 sampel BTA negatif, menunjukkan hasil positif pada 21 sampel. Besarnya produk first-round dan second-round PCR masing-masing adalah 205 bp dan 157 bp.

Kesimpulan
Nested PCR dengan sequencing lebih sensitif dan spesifik dalam mendeteksi  M. tuberculosis dan resistensinya terhadap rifampisin.

Article Details

How to Cite
Rosilawati, M. L. (2007). Deteksi Mycobacterium tuberculosis dan resistensinya terhadap rifampisin dengan Metode nested polymerase chain reaction (PCR) dan sequencing. Universa Medicina, 26(1), 1–10. https://doi.org/10.18051/UnivMed.2007.v26.1-10
Section
Review Article

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