ANALISIS POLIMORFISME GENE SOLUTE CARRIER FAMILY 2, FACILITATED GLUCOSE TRANSPORTER MEMBER 4 DENGAN TEKNIK PCR-RFLP

Larasati, Putu Diva (2025) ANALISIS POLIMORFISME GENE SOLUTE CARRIER FAMILY 2, FACILITATED GLUCOSE TRANSPORTER MEMBER 4 DENGAN TEKNIK PCR-RFLP. Diploma thesis, Poltekkes Kemenkes Denpasar Jurusan Teknologi Laboratorium Medis 2025.

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Abstract

ANALYSIS OF SOLUTE CARRIER FAMILY 2, FACILITATED GLUCOSE TRANSPORTER MEMBER 4 GENE POLYMORPHISM USING PCR RFLP TECHNIQUE ABSTRACT Background: The SLC2A4 gene encodes the GLUT4 protein that plays an important role in insulin-triggered glucose uptake. Polymorphisms of this gene may affect GLUT4 expression and function, thus contributing to insulin resistance and the pathogenesis of type II diabetes mellitus. The aim of the study was to detect SLC2A4 gene polymorphisms in type II DM and non-DM patients using specially designed SLC2A4 gene-specific primers containing EcoRI recognition sites by PCR-RFLP technique. Methods: descriptive study on 20 blood samples (10 type II DM, 10 non-DM) obtained by purposive sampling at Puskesmas II North Denpasar and Denpasar Polytechnic Laboratory. DNA was extracted with Wizard® Genomic DNA Purification Kit (Promega), amplified with forward primer 5′ CTCATTCCCCTCGTGTGACTCT-3′ and reverse primer 5′ ATACATTTAGTACAACCGACCTTTGGT-3′, EcoRI digested, and separated on 1.5% agarose. Results: univariate analysis showed that in the DM group 70% of samples were homozygous wild type (712 bp and 288 bp) and 30% showed no bands, while in the non-DM group 90% were homozygous wild type, while 10% were heterozygous (1,000 bp, 712 bp, 288 bp). In conclusion, the PCR-RFLP technique with SLC2A4 gene primers containing EcoRI recognition sites is able to detect SLC2A4 gene polymorphisms. The homozygous wild type genotype was dominant in both groups and the heterozygous variant was detected in only one non DM sample. Keywords: SLC2A4, GLUT4, gene polymorphism, PCR–RFLP ANALISIS POLIMORFISME GENE SOLUTE CARRIER FAMILY 2, FACILITATED GLUCOSE TRANSPORTER MEMBER 4 DENGAN TEKNIK PCR-RFLP ABSTRAK Latar Belakang: Gen SLC2A4 mengkode protein GLUT4 yang berperan penting dalam penyerapan glukosa yang dipicu insulin. Polimorfisme gen ini dapat memengaruhi ekspresi dan fungsi GLUT4, sehingga berkontribusi pada resistensi insulin dan patogenesis diabetes melitus tipe II. Tujuan penelitian untuk mendeteksi polimorfisme gen SLC2A4 pada pasien DM tipe II dan non-DM dengan menggunakan primer spesifik gen SLC2A4 yang dirancang khusus mengandung situs pengenalan EcoRI dengan teknik PCR-RFLP. Metode: penelitian deskriptif pada 20 sampel darah (10 DM tipe II, 10 non-DM) yang diperoleh secara purposive sampling di Puskesmas II Denpasar Utara dan Laboratorium Poltekkes Denpasar. DNA diekstraksi dengan Wizard® Genomic DNA Purification Kit (Promega), diamplifikasi dengan primer forward 5′-CTCATTCCCCTCGTGTGACTCT-3′ dan primer reverse 5′-ATACATTTAGTACAACCGACCTTTGGT-3′, dicerna EcoRI, dan dipisahkan pada agarosa 1,5%. Hasil: analisis univariat menunjukkan bahwa pada kelompok DM 70% sampel homozigot wild type (712 bp dan 288 bp) dan 30% tidak menunjukkan pita, sedangkan pada kelompok non-DM 90 % homozigot wild type, sementara 10% heterozigot (1.000 bp, 712 bp, 288 bp). Kesimpulannya, teknik PCR–RFLP dengan primer gen SLC2A4 yang mengandung situs pengenalan EcoRI mampu menderteksi polimorfisme gen SLC2A4. Genotipe homozigot wild type dominan pada kedua kelompok dan varian heterozigot terdeteksi hanya ditemukan pada satu sampel non-DM. Kata kunci: SLC2A4, GLUT4, polimorfisme gen, PCR–RFLP

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