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All Journal Medical Laboratory Technology Journal Biosaintifika: Journal of Biology & Biology Education
Intan Razari
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Health Professions Immunology & microbiology Nursing Public Health

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The Use of High Resolution Melting (HRM) Method to Detect rs1800629 of Tumor Necrosis Factor-alpha (TNF-alpha) Gene among Tuberculosis Patients Kinasih Prayuni; Intan Razari; Silviatun Nihayah; Rika Yuliwulandari
Medical Laboratory Technology Journal Vol. 7 No. 1 (2021): June
Publisher : Poltekkes Kemenkes Banjarmasin Jurusan Analis Kesehatan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (439.767 KB) | DOI: 10.31964/mltj.v7i1.362

Abstract

The rs1800629 polymorphism plays a crucial role in the pathogenesis of infectious and autoimmune diseases. Meanwhile, tuberculosis (TB) remains a health primary infectious disease in Indonesia. The purpose of this study is to evaluate the HRM method in detecting the rs1800629 genotype, in the TNF-α gene’s promoter region, within TB patients. The benefit of this study is to accelerate the detection of rs1800629 with a simple, rapid, and cost-effective method for genotyping and mutation screening that does not include the use of a fluorescent probe. In this experimental study, the rs1800629 genotyped in a total of 25 tuberculosis patients using KAPA HRM kit in MyGo Mini PCR, and all amplified PCR products subsequently dispatched for direct DNA sequencing to Macrogen Inc, South Korea. Based on the results, a 100% concordance find in the genotyping of rs1800629 between HRM and sequencing. The authors provided evidence to use HRM in detecting rs1800629 within the TNF-α promotor region. This application as a genotyping assay in tuberculosis patients is a low-cost, rapid, and accurate detection. However, further studies using the HRM method in case-control samples of tuberculosis are required to evaluate the method’s effectiveness and to obtain more information regarding the genotype’s susceptibility to tuberculosis and its adverse effect treatment, including anti-tuberculosis drug, induced liver injury (AT-DILI), and multidrug-resistant TB (MDR-TB), within the Indonesian population.
Correlation of Human Telomerase Reverse Transcriptase Promoter (hTERT) Gene Methylation and Ageing Nihayah, Silviatun; Wening Sari; Yusnita, Yusnita; Intan Razari; Kinasih Prayuni; Utomo, Ahmad Rusdan Handoyo
Biosaintifika: Journal of Biology & Biology Education Vol. 17 No. 2 (2025): August 2025
Publisher : Universitas Negeri Semarang

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15294/biosaintifika.v17i2.22038

Abstract

Promoter methylation of the hTERT gene in blood DNA has been proposed to be an epigenetic molecular clock because it is negatively correlated with ageing. Saliva is an alternative source of DNA because it contains buccal cells. Currently, the correlation of hTERT promoter methylation and ageing using saliva is not known. This study aimed to determine: first, the correlation between hTERT promoter methylation and ageing in saliva, as a source of non-invasive DNA sampling, instead of blood was determined. Second, the influence of sex on the methylation of the hTERT promoter in ageing was evaluated. A cross-sectional study design was used, and 119 subjects were recruited, consisting of 25 children (1-5 y.o), 42 teens (17-19 y.o), 16 adults (20-50 y.o), and 36 elderly (60 to 84 y.o). Promoter methylation of the hTERT of extracted DNA was determined using the MSRE (methyl-specific restriction enzyme) method. The relationship between age and the percentage of hTERT methylation was assessed using the Pearson test. The percentage of hTERT methylation in saliva DNA was negatively correlated with age r= -0.4305 (p-value <0.05). Negative correlation was also found in men (r= -0.376) and women (r=-0.43). Negative correlation between hTERT and ageing has been confirmed in saliva as a noninvasive sampling method. The benefit of this research in ethical and social considerations may encourage a greater participation in ageing research, particularly in underprivileged communities, thus democratising access to scientific advancements in longevity studies.
Co-Authors Kinasih Prayuni Kinasih Prayuni Nihayah, Silviatun Rika Yuliwulandari Silviatun Nihayah Utomo, Ahmad Rusdan Handoyo Wening Sari YUSNITA YUSNITA