Study on association of HSP70_hom gene polymorphism with rheumatoid arthritis using capillary electrophoresis-laser induced fluorescence

摘要

A method using capillary electrophoresis-laser induced fluorescence (CE-LIF) for HSP70_hom gene polymorphism detection has been developed, which was applied to analyze the correlation between rheumatoid arthritis (RA) and HSP70_hom gene polymorphism. The conditions of CE-LIF detection were optimized using DL-1000 DNA Marker, including the concentration of sieving matrix, separation voltage and the concentration of running buffer. The whole genome DNA was extracted from 101 cases of RA and 58 control subjects, amplified by specific primers for PCR, and then digested by NcoI restriction enzyme. The enzyme-digested products were detected by the optimized CE-LIF method and the results were analyzed by logistic regression. Under the optimized conditions of CE-LIF, DL-1000 DNA Marker was detected successively in the same day and the relative standard deviations (RSDs) of migration time was 0.29%-0.41%. There were 58 cases of TT genotype, 41 cases of TC genotype and 2 cases of CC genotype in the case group; 34 cases of TT genotype, 21 cases of TC genotype and 3 cases of CC genotype in the control group. After adjusted age and gender, no significant difference was observed in the distribution of HSP70_hom gene polymorphism between the case group and the control group (P>0.05). The method of CE-LIF detection had the advantages of rapid analysis, high resolution and environmental friendliness, and could be applied to the detection of gene polymorphism; HSP70_hom gene polymorphism was not significantly associated with rheumatoid arthritis. (C) 2017 Elsevier B.V. All rights reserved.