基因芯片在检测慢性乙型肝炎前C区、C启动子区和P区基因变异中的临床应用研究

Clinical application of gene chips in detection of the pre - core and BCP mutations in chronic hepatitis B patients

目的应用基因芯片检测慢性乙肝患者HBV变异类型及其与乙肝标志物和HBVDNA载量的关系，为合理指导临床对慢性乙肝患者进行抗病毒治疗提供科学依据。方法149倒慢性乙肝患者采用酶联免疫法检测乙肝标志物，采用荧光定量PcR法检测RBVDNA载量，采用基因芯片法检测HBV变异。结果在libV感染的不同模式中，HBV—DNA阳性共检出128例。检出阳性率85．90％（128／149），其中大三阳98．82％（84／85）、小三阳75．75％（28／33）、表面抗原与核心抗体阳性51．61％（16／31）。在三组中前c区突变率分别为14．12％、27．27％和33．33％，1组与2、3组比差别有显著性意义（P〈0．01）；BCP区突变率分剐为47．06％、48．48％和58．06％，1组与3组比剐差别有显著性意义（P〈0．05）；P区突变率分别为82．35％、30．30％和51．61％。1组与其他两组比较差别有显著性意义（P〈0．01）；HBVDNA载量≥106组前C区、BCP区和P区突变率分别为48．68％、58．67％和81．33％。与HBV DNA载量104组比，差别均有显著性意义（P〈0．01）。前C区和P区突变率与HBVDNA载量105组比差别有显著性意义（P〈0．05）。结论基因芯片检测HBV基因变异方法简便，特异性高，变异模式判读直观，结果可靠。在治疗前和治疗过程中检测YMDD变异情况。对指导争调整抗病毒治疗的方案有重要的临床应用价值。

Objective To explore relationship between the mutation patterns of the pie - core and BCP of the hepatitis B virus （HBV） DNA polymerase gene and antiviral treated with lamivudine in chronic hepatitis B patients, in order to guide reasonably antiviral therapy in clinic and predict prognosis of the patients. Methods 149 cases of chronic hepatitis B patients, to detect hepatitis B antigen and antibody with enzyme linked immunosorbent assay, use real time PCR to detect HBV DNA, and YMDD variants were detected by nowly advanced gene chip technique. Results Out of 149 patients, 128 cases were HBV DNA positive （85.90%） ,in which, 84 cases were HBsAg, HBeAg and HBeAb positive （group 1 ）, 28 cases were HBsAg, HBeAb and HBcAb positive （group 2） and 16 cases were HBsAg and HBcAb positive（ group 3 ）. Out of three groups, the rate of Pre - core mutation were 14.12%, 27.27% and 33.33%, the rate of BCP mutation were 47.06%, 48.48% and 58.06%, and the rate of P mutation were 82.35%, 30.30% and 51.61. If the HBV DNA level ≥ 10^6 copies/ml , Pre - C , BCP and P mutation rate were 8.68 % ,58.67 % and 81.33 %. Conclusion C, ene chip assay is simple , reliable and has high specificity. It can directly show the pattern of YMDD mutation. To guide reasonably antiveral therapy in clinic and make therapic remedy, it is necessary for HBV - infected patients before or after antiviral therapy to detect HBV YMDD mutations.