Frequency Variant rs1051740 T337C in Carbamazepine Carbide Enzyme Enzyme (EPHX1) Genes on Healthy Volunteers of Javanese Women in Yogyakarta
Abstract
Carbamazepine (CBZ) is a first-line antiepileptic drug belonging to a class
of drugs with a narrow therapeutic index. Women are having hormonal factors
that can increase seizure seizures. Gen Epoxide Hydrolase 1 (EPHX1) is a
microsomal epoxide hydrolase (mEH) enzyme coding gene for CBZ metabolism
which is reported to have variations in rs1051740 with a Minor Allele Frequency
(MAF) value of 0.3133. This study aims to examine the variance frequency of
rs1051740 T337C gene EPHX1 encoding CBZ metabolism enzyme in healthy
female volunteers of Javanese Tribe in Yogyakarta Special Province province.
Methods of research conducted DNA isolation on blood samples of healthy
volunteers women Javanese as much as 50 subjects test by silica gel method, then
do DNA amplification by using Polymerase Chain Reaction (PCR) method.
Primer used forward-5'GATAAGTTCCGTTTCACCC-3 'and reverse
5'TGTTCCTGCCTAGCTCTAA-3'. PCR conditions were initial denaturation of
94 ° C for 3 minutes followed by 32 cycles of denaturation at 94 ° C for 30
seconds, the annealing temperature of 55 ° C for 25 seconds and extension of 72 °
C for 1 minute and the final extension at 72 ° C for 5 minutes. PCR 368bp product
was optimized for Length Polymorphism Restriction Fragment (RFLP) by using
an EcoRV restriction enzyme, then screening all PCR products to ensure that the
enzyme was not able to cut the target sequence. Continued confirmation using the
Sequencing method in three randomly selected samples. The results of the study
showed that there were no gene variations when done using the RFLP method.
There are two out of three samples of nucleotide sequence confirmed by
Sequencing DNA method is heterozygous (CT). It is recommended to do research
with Real-Time PCR method.
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