限制性片段长度多态性分析法在CXCL10G-201A位点变异检测中的应用

目的 通过检测慢性乙型肝炎重型(chronic severe hepatitis B,CSHB)患者趋化因子CXCL10[干扰素γ诱导蛋白(IFNγ-inducible protein,IP-10)]基因启动子区G-201A位点的变异率,探讨PCR-限制性片段长度多态性(restriction fragment len...

Application of restriction fragment length polymorphism method to detecting the mutation of CXCL10 gene promoter G-201A site

Objective To investigate the application of restriction fragment length polymorphism(RFLP) method to the identi-fication of the volume single nucleotide polymorphism(SNP) genotyping by detecting the mutation rate of chemokine(C-X-C motif) ligand 10(CXCL10),also named as IFN γ-inducible protein(IP-10),gene promoter G-201A site in patients with chronic severe hepatitis B(CSHB).Methods Totally 200 CSHB patients in 302 hospital of PLA(CSHB group) and 300 normal controls(NC group) were included in the study.The whole blood anticoagulated with EDTA-K2 was collected,leukocyte DNA was extracted,and PCR-RFLP method was used to detect polymorphisms.Meanwhile,185 samples including 93(46.5%) from CSHB group and 92(30.7%) from NC group were verified by the method of DNA sequencing.Results The agreement between PCR-RFLP method and direct sequencing analysis was statistically significant(k=0.937,P<0.05).The agreement rate of the two methods(97.84%) was in accordance with professional requirements(≥95%).The mutation rates of G-201A related to chemokine IP-10 was 21.28% in CSHB group and 13.82% in NC group,and the difference between them was significant(P<0.05).Conclusions The mutation rate of chemokine IP-10 gene promoter G-201A site in CSHB group is higher than that in NC group.Our study shows that PCR-RFLP method verified by direct sequencing analysis is reliable,and can be used for the volume SNP genotyping identification.