Deteksi Mycobacterium tuberculosis pada Sputum Penderita Tuberkulosis Paru Basil Tahan Asam Negatif dengan Reaksi Rantai Polimerase

Abstract

Pulmonary tuberculosis is an infectious disease remains a public health problem in Indonesia and around the world. Tuberculosis (Tb) is an infectious disease caused by rod-shaped bacteria (bacilli), known as Mycobacterium tuberculosis in most developing countries, the examination of acid-fast bacilli smear, culture and radiological examination was used as a method of diagnosis of tuberculosis in humans. Advanced techniques for the diagnosis of M. rapid and specific tuberculosis is highly emphasized the need for eradication of tuberculosis. Detection of M. tuberculosis by polymerase chain reaction plays an important role for the diagnosis of smear negative cases, early diagnosis of infection, monitoring response to treatment, to determine drug resistance in patients and confirm the certainty of free M. tuberculosis. The purpose of this study was to detect M. tuberculosis in patients with pulmonary tuberculosis sputum acid-fast bacilli staining (AFB) negative by polymerase chain reaction. A total of 36 samples of sputum smear-negative pulmonary TB patients coming from Haji Adam Malik Hospital, General Hospital dr. Pirngadi and Lung Diseases Clinical Centre Medan examined crosssectional descriptive method. Detection of M. tuberculosis carried out using semi-quantitative PCR techniques SYBR Green I-based gene targeting IS6110 and primers T4 and T5. Sputum samples decontaminated with NALC-NaOH method is then performed by the method of DNA extraction spin columns. From all 36 samples of sputum smear-negative pulmonary TB patients from 25 men and 11 women, aged 17-78 years found a positive PCR test results were 13 samples (36.11%) and negative PCR results of 23 samples (63.88%). PCR can be used to support the diagnosis when the clinical, radiological support pulmonary Tb with negative sputum smear microscopic, so no need to wait for culture results for two months. PCR is also an examination of the very useful, fast and effective anti-TB treatment monitoring in patients and confirm the certainty of free M. tuberculosis. The weakness of the PCR test is able to give positive results even though there are only 1-10 germs or bacteria in the examination of specimens dead / dormant and contamination from other specimens. However, in this study has been carried out of action - preventive action to avoid false positives such as the arrangement and the sterilization room, the discipline of working procedures, selection and sterilization of equipment / materials and methods used.