Description of polymerase chain reaction and sequencing DNA Mycobacterium tuberculosis from specimen sputum of tuberculosis patients in Medan

1st International Conference on Tropical Medicine and Infectious Diseases (ICTROMI) Faculty of Medicine Universitas Sumatera Utara

Lily^1 ; Siregar, Y.^2 ; Ilyas, S.^3

Master of Biomedical Sciences, Faculty of Medicine, Universitas Sumatera Utara, Medan

20155, Indonesia^1

Department of Biomedical Sciences, Faculty of Medicine, Universitas Sumatera Utara, Medan

20155, Indonesia^2

Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara, Medan

20155, Indonesia^3

关键词: Agarose gel electrophoresis;    Base pairs;    M. tuberculosis;    Mycobacterium tuberculosis;    PCR method;    PCR products;    Sequencing analysis;    Tuberculosis patients;   

Others  :  https://iopscience.iop.org/article/10.1088/1755-1315/125/1/012145/pdf
DOI  :  10.1088/1755-1315/125/1/012145

来源: IOP

【 摘 要 】

This study purposed to describe the product Polymerase Chain Reaction (PCR) and sequencing of DNA Mycobacterium (M.) tuberculosis from sputum of tuberculosis (TB) patients in Medan. Sputum was collected from patients that diagnosed with pulmonary TB by a physician. Specimen processed by PCR method of Li et al. and sequencing at Macrogen Laboratory. All of 12 product PCR were showed brightness bands at 126 base pair (bp). These results indicated similarity to the study of Li et al. Sequencing analysis showed the presence of a mutation and non-mutation groups of M. tuberculosis. The reference and outcome berange of the mutation and non-mutation of M. tuberculosis were 56-107, 59-85, 60-120 and 63-94, respectively. The percentage bp difference between the outcome and references for mutation and non-mutation were 3.448-6.569and 3.278-7.428%, respectively. In conclusion, the successful amplification of PCR products in a 1.5% agarose gel electrophoresis where all 12 sputa contained rpoB-positive M. tuberculosis and 0.644% difference was found between the outcome with reference bp of the mutation and non-mutation M. tuberculosis groups.