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Establishing a cost-effective tetra-primer amplification refractory mutation system PCR for genotyping the apolipoprotein-A5 -1131T>C polymorphism: a preliminary study for genetic association analysis in the Jambi Malay population

Proceedings Academic Universitas Jambi · Vol. 1 No. 2 (2025) · hlm. 400–409

DOI
10.22437/proca.v1i2.50262
Terbit
2025-11-26
Penerbit
LPPM Universitas Jambi
Bahasa
en
Lisensi
creativecommons.org/licenses/by/4.0

Abstrak

Background: The Apolipoprotein A5 (APOA5) gene's -1131T>C polymorphism is a significant genetic marker for dyslipidemia, with effects varying across ethnicities. Investigating this in specific populations like the Jambi Malay requires genotyping methods that are both reliable and feasible for large-scale studies, often in limited-resource settings. Methods: DNA was extracted from blood samples using a solid-phase column extraction method and subsequently diluted. Four specific primers (two outer and two inner allele-specific) for the T-ARMS PCR assay were designed. The PCR protocol was optimized by varying the annealing temperature, while other parameters like initial denaturation (95°C) and extension (72°C) were kept constant based on standard literature. Amplified products were visualized using gel electrophoresis. Results: An optimized T-ARMS PCR protocol was successfully established. The assay produced clear and distinct banding patterns on a 2.0% agarose gel, allowing for unambiguous genotype calling. Homozygous wild-type (TT) samples showed two bands (512 bp control and 355 bp T-allele), homozygous variant (CC) samples showed two bands (512 bp control and 194 bp C-allele), and heterozygous (TC) samples showed three bands (512 bp, 355 bp, and 194 bp). The assay demonstrated high specificity with no non-specific amplification at an optimal annealing temperature of 59°C. Conclusion: The developed T-ARMS PCR assay is a robust, cost-effective, and reliable method for genotyping the APOA5 rs662799 polymorphism. This tool is well-suited for large-scale molecular epidemiological studies to investigate the genetic architecture of dyslipidemia in the Jambi Malay and other resource-limited settings.

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  • Universitas Jambi
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Rina Nofri EnisAfiliasi tidak diketahui29
Nyimas Natasha Ayu ShafiraAfiliasi tidak diketahui27
Amelia Dwi FitriAfiliasi tidak diketahui23
Susan TarawifaAfiliasi tidak diketahui15
Erny KusdiyahAfiliasi tidak diketahui15
Herlambang HerlambangUniversitas Jambi14
Rita HalimAfiliasi tidak diketahui14
HafizahAfiliasi tidak diketahui12
Wahyu Indah Dewi AuroraAfiliasi tidak diketahui10
Esa Indah AyudiaAfiliasi tidak diketahui9
Zahra Frizki AstyAfiliasi tidak diketahui9
Zilzikridini WijayantiAfiliasi tidak diketahui9
MiftahurrahmahAfiliasi tidak diketahui6
Ahmad SyauqyAfiliasi tidak diketahui6
Tia Wida Ekaputri HzAfiliasi tidak diketahui5
Tia Wida EkaputriAfiliasi tidak diketahui5
ELFIANI ELFIANIAfiliasi tidak diketahui4
Mara Imam Taufiq SiregarAfiliasi tidak diketahui4
HerlambangAfiliasi tidak diketahui4
Fathnur Sani KasmadiAfiliasi tidak diketahui3
Andi SubandiAfiliasi tidak diketahui3
FairuzAfiliasi tidak diketahui2
HumaryantoAfiliasi tidak diketahui2
Mirna Marhami IskandarAfiliasi tidak diketahui2
Armaidi DarmawanAfiliasi tidak diketahui2
Hasna DewiAfiliasi tidak diketahui2
HaninaAfiliasi tidak diketahui2
Patrick Wiliam GadingAfiliasi tidak diketahui2
Charles Apul SimanjuntakAfiliasi tidak diketahui1
Nys. Natasha Ayu SAfiliasi tidak diketahui1

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Sumber metadata: OAI-PMH (metadata jurnal) https://online-journal.unja.ac.id/index.php/PROCA/oai, pertama di-harvest 6 Nov 2026, terakhir terlihat 8 Nov 2026.
Metadata asli (oai_dc) 
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	<dc:title xml:lang="en">Establishing a cost-effective tetra-primer amplification refractory mutation system PCR for genotyping the apolipoprotein-A5 -1131T&gt;C polymorphism: a preliminary study for genetic association analysis in the Jambi Malay population</dc:title>
	<dc:creator xml:lang="en">Perkasa, Tengku Arief Buana</dc:creator>
	<dc:creator xml:lang="en">Amatullah, Afifah</dc:creator>
	<dc:creator xml:lang="en">Hardiningsih, Denok Tri</dc:creator>
	<dc:creator xml:lang="en">Delfira, Annissa</dc:creator>
	<dc:creator xml:lang="en">Puspasari, Anggelia</dc:creator>
	<dc:creator xml:lang="en">Maharani, Citra</dc:creator>
	<dc:creator xml:lang="en">Tarigan, Nadir Putra Indra</dc:creator>
	<dc:creator xml:lang="en">Arrahma, Widya Lawra</dc:creator>
	<dc:creator xml:lang="en">Juanda, Syahreza Hadi</dc:creator>
	<dc:creator xml:lang="en">Buana, Tengku Irfan Wira</dc:creator>
	<dc:subject xml:lang="en">APOA5 -1131T&gt;C Polymorphism</dc:subject>
	<dc:subject xml:lang="en">T-ARMS PCR</dc:subject>
	<dc:subject xml:lang="en">Genotyping</dc:subject>
	<dc:subject xml:lang="en">Annealing Temperature</dc:subject>
	<dc:subject xml:lang="en">Optimization</dc:subject>
	<dc:subject xml:lang="en">Cost-Effective</dc:subject>
	<dc:subject xml:lang="en">Jambi Malay</dc:subject>
	<dc:description xml:lang="en">Background: The Apolipoprotein A5 (APOA5) gene's -1131T&amp;gt;C polymorphism is a significant genetic marker for dyslipidemia, with effects varying across ethnicities. Investigating this in specific populations like the Jambi Malay requires genotyping methods that are both reliable and feasible for large-scale studies, often in limited-resource settings. Methods: DNA was extracted from blood samples using a solid-phase column extraction method and subsequently diluted. Four specific primers (two outer and two inner allele-specific) for the T-ARMS PCR assay were designed. The PCR protocol was optimized by varying the annealing temperature, while other parameters like initial denaturation (95°C) and extension (72°C) were kept constant based on standard literature. Amplified products were visualized using gel electrophoresis. Results: An optimized T-ARMS PCR protocol was successfully established. The assay produced clear and distinct banding patterns on a 2.0% agarose gel, allowing for unambiguous genotype calling. Homozygous wild-type (TT) samples showed two bands (512 bp control and 355 bp T-allele), homozygous variant (CC) samples showed two bands (512 bp control and 194 bp C-allele), and heterozygous (TC) samples showed three bands (512 bp, 355 bp, and 194 bp). The assay demonstrated high specificity with no non-specific amplification at an optimal annealing temperature of 59°C. Conclusion: The developed T-ARMS PCR assay is a robust, cost-effective, and reliable method for genotyping the APOA5 rs662799 polymorphism. This tool is well-suited for large-scale molecular epidemiological studies to investigate the genetic architecture of dyslipidemia in the Jambi Malay and other resource-limited settings.</dc:description>
	<dc:publisher xml:lang="en">LPPM Universitas Jambi</dc:publisher>
	<dc:date>2025-11-26</dc:date>
	<dc:type>info:eu-repo/semantics/article</dc:type>
	<dc:type>info:eu-repo/semantics/publishedVersion</dc:type>
	<dc:format>application/pdf</dc:format>
	<dc:identifier>https://online-journal.unja.ac.id/PROCA/article/view/50262</dc:identifier>
	<dc:identifier>10.22437/proca.v1i2.50262</dc:identifier>
	<dc:source xml:lang="en">Proceedings Academic Universitas Jambi; Vol. 1 No. 2 (2025): The 6th Jambi Medicine and Health Science International Conference (JAMHESIC); 400-409</dc:source>
	<dc:source>10.22437/proca.v1i2</dc:source>
	<dc:language>en</dc:language>
	<dc:relation>https://online-journal.unja.ac.id/PROCA/article/view/50262/22595</dc:relation>
	<dc:rights xml:lang="en">Copyright (c) 2025 Tengku Arief Buana Perkasa, Afifah Amatullah, Denok Tri Hardiningsih, Annissa Delfira, Anggelia Puspasari, Citra Maharani, Nadir Putra Indra Tarigan, Widya Lawra Arrahma, Syahreza Hadi Juanda, Tengku Irfan Wira Buana</dc:rights>
	<dc:rights xml:lang="en">https://creativecommons.org/licenses/by/4.0</dc:rights>
</oai_dc:dc>