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Development of a Gelatin-Based Genomic Reference Material for Halal Authentication Using Real-Time PCR Rahma, Anisa Aula; Meilani, Nanda Diva; Sulistiawati; Ainaputri, Aliza Salsabila; Damara, Dandy Satria; Malau, Jekmal
Science and Technology Indonesia Vol. 10 No. 1 (2025): January
Publisher : Research Center of Inorganic Materials and Coordination Complexes, FMIPA Universitas Sriwijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26554/sti.2025.10.1.27-42

Abstract

Indonesia, home to over 270 million people, has the largest Muslim population globally, with approximately 87.18% adhering to Islam, driving significant demand for halal products, particularly in the food and pharmaceutical sectors. Gelatin, commonly used in medicinal capsules, often originates from porcine sources, necessitating precise halal authentication methods. This study presents the development of a novel genomic DNA-based Reference Material (RM) for gelatin, specifically for porcine DNA detection, employing Real-Time Polymerase Chain Reaction (qPCR) techniques. The methodology encompassed in-silico primer design, sample extraction optimization, DNA quality and quantity analysis, linearity assessment, limit of detection (LoD) and quantification (LoQ) determination, and RM characterization. Results indicated that the designed primers could reliably and efficiently detect porcine DNA, with optimal annealing at 58°C and primer concentration at 500 nM, achieving a PCR efficiency of 96.74%. The LoD and LoQ for pork meat samples were determined to be 0.02 pg/uL and 0.004 pg/uL, respectively, while the LoD for porcine gelatin was 0.27 ng/uL. The RMs exhibited robust homogeneity (Sig. 0.052), significant intergroup differences (Sig. 0.000), and low variation (CV 0.96%). Short-term storage at -80°C and -20°C preserved Ct value stability and consistency. Conclusively, this study successfully developed a novel gelatin-based genomic DNA RM for halal authentication, offering a scientifically validated tool that strengthens the halal assurance system, addressing Indonesian consumers’ demand for porcine-free products. These findings hold substantial implications for regulatory authorities, especially in Indonesia, and could inform the development of standardized qPCR RMs for porcine DNA detection in halal compliance testing.
Primer Design of Porcine DNA using Mitochondrial DNA (Cyt b Gene) for Halal Authentication using Polymerase Chain Reaction (PCR) Rohmah, Siti; Irwansyah, Silvana Lestari; Kasasiah, Ahsanal; Malau, Jekmal
Jurnal Sains dan Kesehatan Vol. 7 No. 1 (2025): J. Sains Kes.
Publisher : Fakultas Farmasi, Universitas Mulawarman, Samarinda, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.25026/jsk.v7i1.2418

Abstract

The Muslim community has difficulties in determining the halal status of animal-derived items and their derivatives due to their extensive prevalence. Islamic customers are increasingly becoming more selective and are demanding halal certification for culinary goods. Developing a way to identify porcine DNA in goods is of utmost importance. The Cyt b gene is being used to evaluate swine DNA samples by using primer candidates. The process of generating primers utilizing bioinformatics tools, such as NCBI, MegaXI, Primer3Plus, SnapGene Viewer, and Net Primer websites, is utilized to assess and analyze the effectiveness of laboratory research. The study concluded that the primers effectively amplified pig DNA, but they were unsuccessful in amplifying chicken or beef DNA. After conducting in silico experiments, a total of 7 possible primers were generated. The most advantageous pair of primers was identified, which includes the forward primer 5'-AACATCCGAAATCACACCC-3' and the reverse primer 5'-AGAATGATATTTGTCCTCAGGG-3'. The efficacy of these primers was evaluated in a controlled laboratory environment. Results from the laboratory experiments demonstrate that these particular primers have the ability to amplify the Cyt b gene from the Sus scrofa species at a temperature of 58°C, producing a DNA fragment that is 415 base pairs long. DNA sequencing is essential to verify that the amplified DNA band matches to the Sus scrofa Cyt b gene. Keywords:          PCR, Porcine, Primer, Cyt b, Halal
In Silico Analysis of Actin Gene as a Candidate for DNA Non-Halal Detection Base on Real-Time PCR Waluyo, Seagames; Malau, Jekmal; Raekiansyah, Muhareva; Yulian, Edwin; Hardiman, Imam
Indonesian Journal of Halal Research Vol. 3 No. 2 (2021): August
Publisher : UIN Sunan Gunung Djati Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15575/ijhar.v3i2.12123

Abstract

Actin genes are genes that are common in organisms, and their expression is constitutive. These genes are used for gene normalization and internal control of DNA extraction, but the actin gene is not widely used for halal certification tests. Bioinformatic studies help to analyze the experiment through in silico more deeply before the experiment is carried out in laboratory, making it more efficient and time effective. uMelt is an analysis to predict the melting curve of target amplification in real-time PCR. Real-time PCR has been widely used for screening and detection of pork content in a product. This research aimed to explore actin gene as a candidate for testing pork using qPCR. The study was carried out in two main stages, namely alignment of the DNA sequence and analysis of the melting curve using the uMelt approach. The results showed a set of actin genes containing conserved regions that can be used as degenerate primers with different family-type coverages. Melting curve prediction with uMelt shows differences in tm peaks so as the types of samples can be easily identified. The use of bioinformatic applications such as uMelt helps in the simulation of predicting the melting curve to increase the precision of the analysis.
Increasing Understanding of Medicines in the Community Through the Implementation of DAGUSIBU as a Prevention of Antibiotic Resistance in Pasir Jengkol Village, Karawang Regency Zahra, Aliya Azkia; Malau, Jekmal; Kasasiah, Ahsanal; Ratnasari, Devi; Septiani, Dia; Sholih, Mally Ghinan
Jurnal Kreativitas Pengabdian Kepada Masyarakat (PKM) Vol 8, No 5 (2025): Volume 8 No 5 (2025)
Publisher : Universitas Malahayati Lampung

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33024/jkpm.v8i5.18936

Abstract

ABSTRACT Antibiotic resistance occurs when microorganisms become unresponsive to antibiotic treatment, rendering common therapies ineffective against infections. As a result, the infection becomes increasingly challenging to treat, posing a heightened risk of severe disease progression and increasing the risk of spreading the disease. The causes of the emergence of antibiotic resistance include a number of factors, including excessive use of antibiotics from self-medication, to inappropriate disposal of antibiotic drugs. Karawang Regency, a region in West Java which consists of 10 sub-districts and has a population of more than 2.4 million people. Research conducted in Karawang shows the potential for antibiotic resistance based on use and the environment. It is known that the people of Karawang Regency have poor knowledge and perception regarding antibiotic resistance and use, one of which is Pasir Jengkol village. Therefore, it is necessary to increase understanding of medicines among the people of Karawang Regency, including by socializing the DAGUSIBU movement (Dapatkan, Gunakan, Simpan dan Buang Obat) or Proper Use, Storage and Disposal of Medications and its implementation as a measure to prevent antibiotic resistance. The output of this activity is primarily an increase in understanding of public knowledge through questionnaires which are analyzed using statistics. The results obtained were then tested using a statistical test showing a significant influence (Sig 0.002) increase in knowledge through the difference in pre-test and post-test scores. This outreach activity, which provides educational materials on preventing resistance, has positively impacted the Pasir Jengkol community's knowledge. Therefore, this socialization initiative can be replicated in other areas, particularly in Karawang Regency. Keywords: Antibiotic,  Drug Resistance, DAGUSIBU, Karawang.
Development of Species-Specific Primers Targeting Mitochondrial Cyt b Gene for Porcine DNA Detection in Halal Authentication via Polymerase Chain Reaction (PCR) Malau, Jekmal; Kasasiah, Ahsanal; Zahra, Aliya Azkia; Irwansyah, Silvana Lestari; Siboro, Dewi Pratiwi Purba
JURNAL PEMBELAJARAN DAN BIOLOGI NUKLEUS Vol 11, No 2: Jurnal Pembelajaran Dan Biologi Nukleus June 2025
Publisher : Universitas Labuhanbatu

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.36987/jpbn.v11i2.6799

Abstract

Background: The detection of porcine DNA is critical to ensuring adherence to halal standards, particularly in food and pharmaceutical products. This study aimed to design and validate species-specific primers targeting the mitochondrial Cyt b gene of Sus scrofa for porcine DNA identification. Using in silico tools such as NCBI, Primer3Plus, SnapGene, Mega and NetPrimer, four primer pairs were designed and assessed for specificity and efficiency. Methodology: Laboratory validation involved PCR amplification and bi-directional Sanger sequencing. Findings: The findings demonstrated that primers 2F/2R and 3F/3R successfully amplified the target DNA, producing amplicons of 514 bp and 456 bp, respectively. The primers exhibited high specificity, with no amplification observed in non-target DNA samples, including chicken and beef. Sanger sequencing confirmed that the amplified products corresponded to the Cyt b gene sequence of Sus scrofa with 100 % similarity, as validated through BLAST analysis. This study presents an accurate and dependable molecular method for detecting porcine DNA, with valuable applications in halal authentication and molecular diagnostics. Contribution: The developed primers offer an effective tool for accurately identifying porcine-derived components, addressing the critical demand for species-specific DNA detection to support halal compliance.
Advancements in Real-Time PCR Technologies: A Comprehensive Review of Probe-Based and Non-Probe-Based Assays for Molecular Diagnostics Malau, Jekmal; Zahro, Aurora Fatimatuz; Zahra, Aliya Azkia; Kasasiah, Ahsanal; Meilani, Nanda Diva; Damara, Dandy Satria; Lestari, Agatha Nabilla; Saryono; Wahyono, Daniel Joko
Science and Technology Indonesia Vol. 10 No. 3 (2025): July
Publisher : Research Center of Inorganic Materials and Coordination Complexes, FMIPA Universitas Sriwijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26554/sti.2025.10.3.660-677

Abstract

The decision between probe-based and non-probe-based qPCR assays is crucial, influenced by diagnostic goals and sample characteristics. This review provides an in-depth evaluation of these two assay types, analyzing their principles, strengths, drawbacks, and applications. A thorough review of the literature, primarily sourced from PubMed, was undertaken to explore prominent assay systems, including TaqMan, KASP, rhAmp, HRM, and SYBR Green. Probe-based qPCR assays, exemplified by TaqMan and rhAmp, are distinguished by their high specificity, aptitude for multiplex analysis, and reduced risk of false positives, making them highly suitable for SNP genotyping and pathogen detection. However, their elevated costs and intricate design requirements remain significant challenges. Conversely, non-probe-based assays, such as SYBR Green and HRM, present cost-effective alternatives with straightforward designs. HRM, in particular, is effective in identifying genetic variations like SNPs with remarkable sensitivity. Nonetheless, these methods are susceptible to non-specific amplifications, requiring careful optimization to maintain reliability. The selection of a suitable qPCR assay depends on various factors, including precision, affordability, and multiplexing capabilities, with applications spanning infectious disease detection and genetic disorder analysis. This review emphasizes the indispensable role of qPCR in molecular diagnostics while showcasing recent technological advances that aim to mitigate existing constraints and enhance diagnostic precision and accessibility.
Potensi Aktivitas Antibakteri dan Antioksidan dari Senyawa Bioaktif Bakteri Tanah Asal Samarinda, Kalimantan Timur: The Antibacterial and Antioxidant Potential of Bioactive Metabolites from Soil-Derived Bacteria in Samarinda, East Kalimantan Atwita, Syelziva Yonda; Malau, Jekmal; Permatasari, Vera; Primahana, Gian; Dewijanti, Indah D.; Yuswan, Apriza; Prastya, Muhammad Eka
Jurnal Sumberdaya Hayati Vol. 11 No. 2 (2025): 2025
Publisher : Departemen Biologi, Institut Pertanian Bogor

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/jsdh.11.2.63-73

Abstract

Antibiotic resistance is a global challenge, especially in the treatment of bacterial infections. This study aims to explore the antibacterial and antioxidant potential of bioactive compounds isolated from soil bacteria in Samarinda, East Kalimantan. Of the 30 isolates tested, 1 potential isolate was obtained, namely isolate code T1.16. This potential bacterial isolate was further cultured and fermented in Tryptic Soybean and Luria Bertani Broth medium to obtain its crude secondary metabolite. Interestingly, its crude extract showed antibacterial activity with MIC values of 100.4-3,211 µg/ml against Escherichia coli strain ATCC 8739, Pseudomonas aeruginosa strain ATCC 9027, Bacillus subtilis strain ATCC 6633, and Staphylococcus aureus strain ATCC 25923. Based on 1,1-diphenyl-2-picrylhydrazyl (DPPH) method was used to identify antioxidant activity, IC50 values obtained ranged from 990-1730 µg/ml. Ultimately, by using Gas Chromatography Mass Spectrometry (GC-MS) analysis, bioactive compounds in the form of Cyclo(L-prolyl-L-valine), 2-methylpropyl and phenylmethyl in bacterial extract T1.16 have potential antioxidant and antibacterial bioactivities. These results support the potential development of new antibacterial and antioxidant compounds from soil bacteria, as well as the importance of further exploration for pharmacological applications.