Kemunculan Gen Mycobacterium tuberculosis dari Sampel Darah Pasien Pengobatan Tuberkulosis Fase Konversi Menggunakan PCR yang Tidak Terdeteksi dengan Evaluasi Pewarnaan Bakteri di Puskesmas Janti

Abstract

The conversion phase in tuberculosis treatment examines patients to ensure the possibility of further advancement. Despite its low specificity, microscopic evaluation is a standard method at public health centers. Moreover, the biased results from this method form a new problem related to multidrug-resistant tuberculosis (MDR-TB). Molecular approaches are known for their high specificity and accuracy in microbial identification. Therefore, this study aimed to compare the specificity and sensitivity of molecular and microscopic methods. The sample used was 17 blood of tuberculosis patients in the conversion phase accompanied by one positive and negative control collected from October to November 2020. The DNA sequence was 16s RNA with forwarding ACT GAG ATA CGG CCC AGA CT and reverse primer TCA CGA ACA ACG CGA CAA AC. Furthermore, the research was conducted by optimizing variations in the number of cycles, temperature melting (Tm), and template DNA concentration. The results showed 279 bp length of PCR product, under the optimum conditions under 35 cycles, Tm of 60˚C, and template DNA concentration of 5 µmol. The PCR result identified that 2 of 17 samples contain positive Mycobacterium tuberculosis. Meanwhile, none was positive on microscopic evaluation. Therefore, there are different results between microscopic and molecular methods.