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Genetic Variation Analysis of katG Gene Sequence in Mycobacterium tuberculosis Using RFLP In Silico Nurfalinda, Siti; Aulia, Ony Nattasha; Sirwati, Fadila; Alridho, Rezeki Rival; Achayar, Afifatul
Jurnal Serambi Biologi Vol. 9 No. 1 (2024): Jurnal Serambi Biologi
Publisher : Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Negeri Padang

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24036/srmb.v9i1.337

Abstract

Tuberculosis (TB) is an infectious disease that is the leading cause of death in Indonesia. This infection is caused by Mycobacterium tuberculosis. In Indonesia in 2021, there were approximately 824,000 cases of TB. A total of 8,268 TB cases were associated with resistance of M. tuberculosis to anti-tuberculosis drugs (OAT). MDR-TB is a strain that is resistant to two of the most important anti-tuberculosis drugs in first-line TB therapy, namely Rifampicin (RIF) and Isoniazid (INH). The most common mechanism of INH resistance is the katG gene mutation, found (95%) at codon 315. In the katG gene there is a change in the amino acid Ser (AGC) → Thr (ACC). Genetic variation in a population will affect the survival ability of an individual. This study used BamHI restriction enzyme. This study aims to analyze genetic variation in the KatG gene sequence in M. tuberculosis NCBI popset 2363230696 by using RFLP in silico. The results showed that there were genetic variations in the KatG gene sequence and two allelic variations contained in 7 KatG gene sequences using the BamHI restriction enzyme. Key words : M. tuberculosis, BamHI, KatG , Genetic Variation, RFLP in silico.
Development and Optimization of SARS-CoV-2-Specific Primers for Accurate Diagnosis: A Case Study in West Sumatra - Indonesia Aulia, Ony Nattasha; Putri, Dwi Hilda; Faizal, Irvan
Althea Medical Journal Vol 11, No 4 (2024)
Publisher : Faculty of Medicine Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15850/amj.v11n4.3348

Abstract

Background: In 2022, new cases of Covid-19 emerged, including the Omicron variant which is classified as a variant of concern (VOC). West Sumatra is one of the top ten provinces with the highest number of cases in Indonesia. This study aimed to design specific primers and optimize the PCR method that can be used for accurate detection, specifically for SARS-CoV-2 circulating in West Sumatra.Methods: This study used an in silico approach, using whole genome sequencing (WGS) data available at the global initiative on sharing avian influenza data (GISAID), and employing the Geneious Prime application which confirmed samples collected from Padang, West Sumatra, and from Jakarta, Bogor, Depok, Tangerang, Bekasi (Jabodetabek) serving as comparative sample tests. Technology development was supported by bioinformatics testing, laboratory testing, and validation methods, involving gene mining, sequence alignment, and primer design. Laboratory tests and validation included viral genomes extraction and cDNA synthesis, polymerase chain reaction (PCR) testing, and results analysis. Results: Three sets of optimal primer candidates amplified the coveted target gene was discovered, specifically, the S gene of the receptor binding domain (RBD) region.Conclusions: The primers designed through a consensus between the complete genome of the SARS-CoV-2 isolate Wuhan-Hu-1 and the WGS of the Omicron variant in Padang, West Sumatra, have successfully detected the SARS-CoV-2 virus variant in the region. The most effective temperature optimization results were achieved by testing three primer products on samples from Padang and Jabodetabek. It has significance as a valuable diagnostic tool in the primer form.