XÂY DỰNG QUY TRÌNH ASO PCR XÁC ĐỊNH ĐIỂM ĐA HÌNH ĐƠN R1628P TRÊN GEN LRRK2

Abstract

Background: Parkinson's disease (PD) is among most common neurodegenerative disorders characterized by distinctive motor symptoms of tremor, rigidity, and reduced movement, caused by the progressive loss of dopaminergic neurons in the substantia nigra. The R1628P variant in the LRRK2 gene is a significant risk factor for PD in Asian populations, particularly among Chinese and Thai individuals. Research on R1628P provides further insights into the pathogenesis of PD. Objectives: This study aimed to develop and optimize an ASO-PCR protocol for detecting the R1628P variant in the LRRK2 gene, and to investigate the frequency of this variant in population without Parkinson's disease. Methods: This study was performed using 200 pre-collected DNA samples at the Center for Molecular Biology - University of Medicine and Pharmacy, HCMC. The ASO-PCR process utilized allele-specific primers for G and C alleles of the R1628P variant, with optimized temperature parameters to enhance sensitivity and specificity. Results: The ASO-PCR protocol demonstrated high specificity in genotype detection, with a frequency of the R1628P variant of 3.5%, including 7 samples with G/C genotype and 193 samples with G/G genotype. The genotypes identified by ASO-PCR were consistent with those by Sanger sequencing. Conclusion: The ASO-PCR technique is an effective tool for detecting the R1628P variant, providing a highly accurate method to determine allele frequency and genetic risk of PD.