80例乙型、丙型病毒性肝炎血清、组织基因芯片检测分析

Detection of the Serum and Liver Tissues in 80 Patients with in using DNA microarray Technique

研究病毒性肝炎基因芯片的制备，用肝炎基因诊断芯片、病毒抗原抗体检测方法、免疫组织化学法和原位分子杂交法，分别检测乙型肝炎和丙型肝炎血清及肝组织，比较各种方法的优缺点。将带荧光标记的ＰＣＲ产物用点样仪点于玻片介质上，并经过点样处理后制备成基因芯片。收集乙型肝炎血清４０例、肝组织２０例，丙型肝炎血清２０例，健康人血清４０例，乙型肝炎确诊用雅培设备试剂，丙型肝炎检测抗ＨＣＶ及ＨＣＶ一ＲＮＡ，确诊后同健康人血清一起进行双盲混合编组，用基因诊断芯片检测，２０例乙型肝炎肝组织用免疫组织化学和原位分子杂交法，检测 ＨＢｃＡｇ和 ＨＢＶＤＮＡ。同时用基因芯片检测。４０例乙型肝炎病毒标志物阳性血清，基因诊断芯片检测阳性 ３０例，阴性１０例，２０例丙型肝炎血清基因芯片检测阳性８例。４０例健康人血清，乙型和丙型肝炎基因诊断芯片检测均阴性． ２０例肝组织免疫组化发现ＨＢｃＡｇ阳性２０例，原位分子杂交发现ＨＢＶＤＮＡ阳性１８例，基因芯片检测阳性１８例。肝炎基因诊断芯片可以同时检测乙型和丙型肝炎，诊断乙型肝炎血清准确率可达８０％，假阳性率低。诊断丙型肝炎阳性率低，技术有待完善。

To study the preparation of gene chips to diagnose hepatitis. HBVM,HBV- DNA ,antiHCV.HCVRNA were detected in the serum and liver tissues of patients with hepatitis B or C by gene chips, immunocytochemistry and in situ hybridization in order to compare the advantages and disadvantages of these diagnosis methods. DNA microarrays were prepared by spotting fluorescence labeled PCR products of target genes onto specially treated glass slides with robotics. The serum samples of 40 patients with hepatitis B,20 patients with hepatitis C,40 healthy persons and 30 cases liver tissues with hepatitis B were diagnosised by Abbott reagents.antiHCV.HCVRNA. immunocytochemistry and in situ hybridization Then all of 80 serum samples were tested randomly by gene chips. Out of 40 serum samples of patients with hepatitis B tested by gene chips, 30 cases were positive. In the 20 serum samples of patients with hepatitis C tested by gene chips, 8 cases were positive. Out of 20 cases liver tissues, 18 cases HBVDNA were positive by gene chips, 20 cases HBcAg were positive by immunocytochemistry and 18 cases HBVDNA were positive in situ hybridization. All healthy persons tested by gene chip were negative. Hepatitis B and C can be detected by one gene chip. The accuracy of diagnosing hepatitis B in the serum can reach 80%, the pseudopositive ratio by using gene chips was low. But the accuracy of diagnosing hepatitis C was low. The technique need to be improved.