DETEKSI VARIASI GENETIK GEN PENYANDI ENZIM PEMETABOLISME ASAM VALPROAT UGT1A6 rs2070959 A>G PADA RESPONDEN SEHAT SUKU JAWA DI INDONESIA

Abstract

Epilepsy is a seizure disease which has a risk of death three times higher in patients of epilepsy compared to normal people and also has the risk Sudden Unexpected Death in Epilepsy (SUDEP), which causing sudden death due to epilepsy. As many as 70% of people with epilepsy can be controlled with an Antiepileptic drugs (OAE). One of the first line OAE agents in epilepsy theraphy is Valproic Acid (VA). Valproate monitoring is indispensable for its variability occurance as the result of drug level taken and drug dosing between each individual; also monitoring for variability occurance happens by gender, age, and their genetics. The occurence of polymorphism genetic in metabolizing gene of Valproic Acid (UGT1A6) as a gene encoding the glucoronidase enzyme is resulting different drug level in plasma which may effect therapeutic results. This study aimed to determine the frequency of genetic variations in the UGT1A6 rs207959 A>G in Javanese population. The sample used in this study were stored as DNA speciment from 100 healthy respondent who included the inclusion criteria. The analysis was using by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) method with forward primary 5’CTT TAA GGA GAG CAA GTT TGA TG-3’ and reverse primary 5’-CCA CTC GTT GGG AAA AAG TC-3’. The DNA from PCR amplification digested by NsiI enzyme which cut on ATGCA|T sequences from the DNA template resulted wild type AA (128 and 87 bp) of genotypic variant. Frequency variant of UGT1A6 gene rs2070959 A>G can not be shown in this research. Some trials had been executed for optimizing the cutting by increasing amount of enzyme, time and temperature of incubation, also DNA concentration. The failure of digestion process in optimizing PCR-RFLP was confirmed by Direct Sequencing method on two random samples. Sequencing result showed that there were samples that had genotype variants of AG (heteroziygous mutants) which had the results of cuts, 128, 87 and 215 bp. Therefore, further research is needed related to the detection of variations of the UGT1A6 gene rs2070959 A>G using restriction enzymes other then NsiI in Javanese population.