DNA芯片检测肝组织及血清中乙型肝炎病毒DNA的临床研究

The clinical study on the detecting simultaneity HBV DNA in the serum and liver tissuse of patients with DNA microarry technique

目的 研究DNA芯片检测乙型肝炎和肝硬化患者肝组织及血清中乙型肝炎病毒(HBV)DNA的应用价值。方法 用点样仪将聚合酶链反应 (PCR)扩增的HBVDNA探针制成基因芯片。对 15例慢性乙型肝炎病人的血清和肝活检组织 ,99例乙型肝炎后肝硬化肝组织 ,分别用基因芯片、原位分子杂交、免疫组织化学和雅培试剂检测HBVDNA、乙型肝炎核心抗原 (HBcAg)、乙型肝炎表面抗原 (HBsAg)和乙型肝炎e抗原 (HBeAg)。结果 用基因芯片对 15份HBsAg、HBeAg阳性的乙型肝炎患者血清进行检测 ,HBVDNA均阳性 ,阳性率符合率为 10 0 % ;15份肝活检组织标本 ,免疫组化法检测HBcAg阳性 15例 ,原位分子杂交法和基因芯片检测HBVDNA均阳性 14例 ,阳性率 93%。 99份肝炎后肝硬化患者肝组织标本 ,HBcAg阳性 6 7份 ,HBVDNA阳性 5 3份 ,基因芯片检测HBVDNA阳性 46份 ,阳性率分别为 6 9%、88%。 32例HBcAg、HBVDNA阴性的肝组织中 ,基因芯片检测HBVDNA均为阴性。结论 肝炎基因诊断芯片可检测肝组织及血清中HBVDNA ,诊断准确率高 ,假阳性率低。

Objective To study the application value that the DNA microarryies diagnose hepatitis virus DNA in the serum and liver tissues of patients with hepatitis B and posthepatitic cirrhosis. Methods The DNA microarries were prepared by spotting PCR products labeled fluorescence of the targets genes to the surface of glass slides by the robotics. For HBV DNA?HBcAg?HBsAg and HBeAg in the serum samples and liver tissues of 15 patients with hepatitis B and HBV DNA, HBcAg in the liver tissues of the 99 cases patients with posthepatitic cirrhosis, they were detected and diagnosed by Abbott reagents, immunocytochemistry, by in situ molecule hybridization technique and the gene chips. Results 15 cases serum samples with positive expression of HBsAg and HBeAg diagnosed with Abbott reagents, were all positive diagnosed by the gene chips. For the liver tissues of 15 cases patients, there were 15 cases positive expression of HBcAg and 14 cases positive expression of HBV DNA in liver tissues were positive detected respectively by immunocytochemistry, by in situ molecule hybridization technique and by gene chip. In the 99 liver tissues samples of posthepatitic cirrhosis 67 cases expression of HBcAg, 53 cases expression of HBV DNA were positive detected with immunocytochemistey and with in situ molecule hybridization technique, in which 46 cases expression of HBV DNA were positive detected by gene chip. 32 cases of HBcAg and HBV DNA were negative detected by the three test methods.Conclusion In the serums and liver tissues HBV DNA can be detected simultaneity by the gene chip in diagnosis with great accuracy.