O.N. Aulia, D.H. Putri, L. Chaidir, M. Yusuf, K. Kurniawan, I. Faizal
Tuberculosis (TB), caused by Mycobacterium tuberculosis (MTB), remains one of the leading causes of death in Indonesia. The limited diagnostic tools for TB, including Multi-Drug Resistant Tuberculosis (MDR-TB), contribute to the increasing number of cases. MDR-TB refers to TB strains resistant to Anti-Tuberculosis Drugs such as Isoniazid (INH). In response, the WHO has recommended the use of a rapid, equipment-free method called loop-mediated isothermal amplification (LAMP). This study aims to design primers and optimize the LAMP method for the detection of Isoniazid-resistant M. tuberculosis. The research integrates bioinformatics testing, laboratory experiments, and validation methods. The study successfully designed LAMP primers for detecting Isoniazid-resistant comprising three pairs of primers: F3, B3; FIP, BIP; and LF, LB. The optimization of the LAMP method involved temperature and time parameters. Five temperature treatments were tested, revealing an optimum temperature of 70°C. For time optimization, eight treatments were conducted, with an optimum time range of 25-30 minutes. The study concludes that the LAMP method differs from the PCR method due to the higher number of primers employed and LAMP requires only a single isothermal temperature. Additionally, the study achieved optimization of temperature and time for the LAMP method, facilitating the detection of Isoniazid-resistant MTB. © Published under licence by IOP Publishing Ltd.
Department of Biology, Faculty of Mathematics and Sciences, Universitas Negeri Padang, Jl. Prof. Dr. Hamka, Air Tawar Barat., North Padang District, West Sumatra, Padang City, Indonesia; Center for Translational Biomarker Research, Universitas Padjadjaran, West Java, Bandung, 40132, Indonesia; Department of Chemistry, Faculty of Mathematics and Sciences, Universitas Padjadjaran, Indonesia; Centre for Vaccine and Drug Research, National Research and Innovation Agency, Kawasan Puspiptek Serpong, Serpong, 15314, Indonesia; Department of Biotechnology, Faculty of Biotechnology, Atma Jaya Catholic University of Indonesia, Tangerang, Jakarta Selatan, 15345, Indonesia
