摘要
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.
出处
《临床肝胆病杂志》
CAS
北大核心
2002年第1期28-30,共3页
Journal of Clinical Hepatology
基金
江苏省科学技术委员会重点科研项目(BS2000028)资金资助
江苏省卫生厅2000年重点科研项目资金资助。