Detection and resistance of Mycobacterium tuberculosis on rifampicin using nested polymerase chain reaction and sequencing method  

Oleh:

Rosilawati, Maria Lina

Jenis: Article from Journal - ilmiah nasional - terakreditasi DIKTI - non-atma jaya
Dalam koleksi: Universa Medicina vol. 26 no. 01 (Jan. 2007), page 01.
Topik: Mycobacterium tuberculosis; rpoß DNA; nested PCR; sequencing

Ketersediaan

Perpustakaan FK Nomor Panggil: U01.K.01 Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

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Isi artikel

Background The rpob (RNA polymerase sub unit b) DNA of Mycobacterim tuberculosis can be specifically amplified by using a nested PCR. The nested PCR linked to DNA sequencing was applied directly to detect M. tuberculosis and determine the rpob gene mutation related with the rifampicin resistance either in clinical isolates or sputa. Methods Samples used in this research were 20 clinical isolates and 30 sputa which were amplified with the region of rpob DNA of M. tuberculosis. DNA of clinical isolates and sputum samples were extracted by means of fenol-kloroform and Boom’s methods, respectively. Sequencing method was only applied for sputum samples. Results Of 20 clinical isolates, 15 isolates were positive for M. tuberculosis with nested PCR, 4 isolates were Mycobacteria other than tuberculosis (MOTT) and 1 isolate was non-Mycobacteria. The nested PCR could detect 21 sputum samples of 30 samples consisted of 25 samples with positif acid fast bacilli (AFB) and 5 samples with negative AFB. First-round and second-round PCR products were 205 bp and 157 bp, respectively. Conclusion Nested PCR in the sequencing was more sensitive and specific to detect M. tuberculosis and it resistance to rifampicin.

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