UTILIZATION OF GENOM DNA FOR NON-INVASIVE DETECTION OF BRCA1 GENE MUTATIONS IN OVARIAN CANCER PATIENTS

• Main Author: Fadiyah, Farizul
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/71344
• Institution: Institut Teknologi Bandung

Ovarian cancer is the seventh most common cause of death in women. This is mainly due to the absence of clear symptoms in ovarian cancer patients, resulting in late detection of ovarian cancer. Mutation detection for ovarian cancer can be done by tracing the presence of mutations in the BRCA1 gene with the main target of genomic DNA in the blood and urine of patients with suspected ovarian cancer. Better diagnostic strategies for detection are needed to improve survival/cure rates for women with ovarian cancer. Screening of ovarian cancer patients shows that mutations in the BRCA1 gene increase the occurrence of ovarian cancer by 40-53%. The most common mutation in the BRCA1 gene is the 185delAG mutation in exon 2. This study aims to trace the 185delAG mutation in exon 2 of the BRCA1 gene in ovarian cancer patients using urine, while blood sample to validate the result. The method for this research is the PCR-RFLP method to detect mutation of BRCA1 gene in genomic DNA. The length of BRCA1 gene exon 2 with 185delAG mutation, which is targeted for PCR-RFLP is 498 bp. The process of detecting the 185delAG mutation in exon 2 of the BRCA1 gene uses the specific restriction enzyme HinfI, which cuts the mutated part. The result of the HinfI restriction enzyme in the 185delAG mutation with a size of 498 bp will be truncated into 2 DNA bands with sizes of 294 bp and 204 bp. Genomic DNA from blood and urine samples were obtained from patients with ovarian cancer (45 samples) and non-ovarian cancer (9 samples). Mutation detection using PCR- RFLP method and validated by Sanger sequencing The results of detection using the RFLP PCR method in this study showed that the detection percentage of the 185delAG mutation in exon 2 of the BRCA1 gene from blood and urine samples of ovarian tumor/cancer patients was 15.55%. Detection result in blood and urine samples using PCR-RFLP method showed that 7 patients out of a total of 45 ovarian tumor/cancer patients have the 185delAG mutation . The result of sequencing in blood and urine samples using Sanger sequencing showed that 7 positive patients had the 185delAG mutation. Based on the results of analysis using PCR-RFLP, the 185delAG mutation in exon 2 of the BRCA1 gene occurs in ovarian tumor/cancer patients with malignant or benign tumor status. The positive detection status of the 185delAG mutation occurred in the age group of 25-55 years. This study succeeded in demonstrating that genomic DNA in urine can be used to detect the 185delAG mutation in exon 2 of the BRCA1 gene.