DETEKSI MUTASI GEN APC BERBASIS HIGH RESOLUTION MELTING PADA SAMPEL KANKER KOLOREKTAL

FAISAL RAMADHAN, . (2026) DETEKSI MUTASI GEN APC BERBASIS HIGH RESOLUTION MELTING PADA SAMPEL KANKER KOLOREKTAL. Sarjana thesis, UNIVERSITAS NEGERI JAKARTA.

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Abstract

Kanker kolorektal merupakan salah satu penyebab utama kematian akibat kanker yang berkaitan dengan akumulasi mutasi genetik, termasuk pada gen Adenomatous Polyposis Coli (APC). High Resolution Melting (HRM) merupakan metode berbasis qPCR yang berpotensi digunakan sebagai metode pre-sequencing untuk mendeteksi variasi genetik sebelum dikonfirmasi menggunakan sekuensing Sanger. Penelitian ini bertujuan mendeskripsikan karakteristik subjek penelitian, mengevaluasi desain, karakterisasi, dan optimasi primer gen APC, menganalisis profil qPCR-HRM, serta mengonfirmasi hasilnya menggunakan sekuensing Sanger. Penelitian dilakukan terhadap 50 sampel jaringan kanker kolorektal dan 10 sampel darah individu sehat. Karakteristik subjek penelitian didominasi oleh laki-laki (58%), kelompok usia ≥60 tahun (50%), stadium lanjut (66%), dan metastasis (62%). Primer yang dirancang menunjukkan spesifisitas terhadap daerah target gen APC dan menghasilkan amplifikasi sesuai ukuran yang diharapkan. Analisis qPCR-HRM berhasil mengamplifikasi seluruh sampel secara spesifik, dengan sebagian besar sampel terkelompok dalam Varian1 dan satu sampel (P27) membentuk Varian2. Sekuensing Sanger menunjukkan bahwa mutasi target c.2635C>T (p.Gln879Ter) tidak ditemukan pada seluruh sampel. Namun, pada sampel P27 teridentifikasi dua variasi nukleotida heterozigot, yaitu c.2523A>G (p.Leu841=) dan c.2623A>G (p.Lys875Glu), dengan variasi c.2623A>G diperkirakan berkontribusi terhadap perubahan profil pelelehan DNA. Penelitian ini menunjukkan bahwa qPCR-HRM efektif sebagai metode pre-sequencing untuk mendeteksi variasi gen APC, tetapi identifikasi variasi nukleotida secara definitif tetap memerlukan konfirmasi menggunakan sekuensing Sanger /***** Colorectal cancer is one of the leading causes of cancer-related mortality and is associated with the accumulation of genetic mutations, including those affecting the Adenomatous Polyposis Coli (APC) gene. High Resolution Melting (HRM) is a qPCR-based method with potential as a pre-sequencing approach for detecting genetic variations prior to confirmation by Sanger sequencing. This study aimed to describe the characteristics of the study subjects, evaluate the design, characterization, and optimization of APC gene primers, analyze qPCR-HRM profiles, and confirm the findings using Sanger sequencing. The study involved 50 colorectal cancer tissue samples and 10 blood samples from healthy individuals. The study subjects were predominantly male (58%), aged ≥60 years (50%), diagnosed at advanced stages (66%), and presented with metastasis (62%). The designed primers showed high specificity for the target region of the APC gene and produced amplicons of the expected size. qPCR-HRM successfully amplified all samples specifically, with most samples classified as Variant 1 and one sample (P27) classified as Variant 2. Sanger sequencing confirmed the absence of the target c.2635C>T (p.Gln879Ter) mutation in all analyzed samples. However, sample P27 contained two heterozygous nucleotide variants, c.2523A>G (p.Leu841=) and c.2623A>G (p.Lys875Glu), with the latter likely contributing to the altered DNA melting profile. These findings indicate that qPCR-HRM is effective as a pre-sequencing method for detecting APC gene variations, although definitive identification of nucleotide variations still requires confirmation by Sanger sequencing

Item Type:	Thesis (Sarjana)
Additional Information:	1). Rizky Priambodo, S.Si., M.Si. ; 2). Adhitya Bayu Perdana, S.Si., M.Epid.
Subjects:	Sains > Sains, Ilmu Pengetahuan Alam Sains > Ilmu Bumi > Biologi Sains > Ilmu Bumi > Genetika
Divisions:	FMIPA > S1 Biologi
Depositing User:	Faisal Ramadhan .
Date Deposited:	12 Aug 2026 01:52
Last Modified:	12 Aug 2026 01:52
URI:	http://repository.unj.ac.id/id/eprint/69257

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