Search

Introduction: Tuberculosis (TB) due to infection of the Mycobacterium tuberculosis has become a concern since this disease has been suffered by most of the world's population and causes death in large numbers. The increasing number of TB patients has increased our vigilance to reduce the spread of this disease. One of the efforts to provide effective treatment for cases of infection by this bacterium is to determine the proper anti-tuberculosis drug. Methods: This study tested M. tuberculosis isolates against several tuberculosis drugs, such as isoniazid, quinolone (ofloxacin and moxifloxacin), kanamycin, capreomycin, bedaquiline, linezolid, and clofazimine in MGIT liquid medium. The isolates used were bacterial stock belonging to the Department of Microbiology, Medical Faculty, Universitas Indonesia. All isolates used had a resistance phenotype to the anti-tuberculosis drug rifampicin determined by the GeneXpert MTB/Rif method. Results: From all the isolates tested, the percentage of anti-tuberculosis resistance was as follows: low-dose isoniazid 70.4%, high-dose isoniazid 66.7%, ofloxacin 9.9%, low-dose moxifloxacin 9.9%, high-dose moxifloxacin 2.5%, and bedaquiline 5.1%. There was no resistance to linezolid and clofazimine among the tested isolates. This study also found that 57 isolates were multi-drug resistant (MDR) strains, six isolates were pre-extensively drug resistance (XDR), and one isolate was an XDR strain. Conclusion: This study presented an overview of the resistance profile of M. tuberculosis to several first-, second-line and new tuberculosis drugs in vitro. The results of this study can be used by stakeholders in the health sector to develop policies for better management of tuberculosis. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis. It is a persistent global health problem with a high mortality rate. Currently, TB is controlled by administering the Bacillus Calmette-Guerin (BCG) vaccine, but the effectiveness of its protection varies among individuals in a population. The pe/ppe gene family comprises a typical group of genes that play a role in avoiding the host immune response and inducing persistent TB infection. Based on in silico analysis, the pe11 gene has estimated immunogenicity and potential as a TB seed vaccine candidate. The pe11 gene from an Indonesian isolate of an M. tuberculosis Beijing strain was amplified by polymerase chain reaction (PCR) and inserted into the mammalian expression vector pcDNA3.1. The recombinant vector pcDNA3.1-pe11 was used to transform Top10 competent Escherichia coli. Clones from the transformation were subjected to colony PCR to confirm the direction of the insert. Sequencing was performed to confirm the correctness of the insert sequence. In this study, the pe11 gene was successfully cloned into the pcDNA3.1 vector in the correct direction to assure PE11 expression. No mutations were found in the pe11 gene insert, compared with the M. tuberculosis H37Rv sequence as the standard. A pcDNA3.1 vector containing the pe11 gene derived from an M. tuberculosis Beijing strain was successfully constructed. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (M. tuberculosis). TB is still a major health problem. The Bacillus Calmette-Guerin (BCG) vaccine is the only one available for TB and is known to confer variable levels of protection. Because of this variability, a new vaccine is needed to control TB. Proteins secreted by M. tuberculosis are known to induce protective immunity. Within the genome of M. tuberculosis, there is a family of proteins called resuscitation promoting factor (Rpf), which plays a role in the reactivation of M. tuberculosis. RpfD is a member of the Rpf family that has been shown to be immunogenic, making it suitable for use as a TB vaccine. The rpfD gene of the M. tuberculosis Beijing strain from the bacterial stock of the Department of Microbiology at the Medical Faculty of the Universitas Indonesia was amplified using polymerase chain reaction (PCR) and then inserted into the mammalian expression vector pcDNA3.1(+). Then, the pcDNA3.1(+)-rpfD vector was transformed to Escherichia coli DH5α. A 465-bp target fragment was obtained, and the accuracy of the cloning was confirmed using colony PCR, restriction enzyme digestion, and sequencing. We expect that this recombinant plasmid will induce immunity in future animal models and thus will prove itself to be a candidate for an M. tuberculosis vaccine. [ABSTRACT FROM AUTHOR]