基因芯片技术检测慢性乙型肝炎肝硬化患者肝组织及血清HBVDNA研究

Study of the gene chips techonlogy in detecting HBVDNA in the serums and liver tissues of patients with posthepatitic cirrhosis

目的研究DNA芯片技术在检测乙型肝炎和肝硬化患者肝组织及血清中HBVDNA的应用,并与雅培试剂、免疫组织化学和原位分子杂交法比较。方法用点样仪将PCR扩增的HBVDNA探针制成基因芯片,对15例慢性乙型肝炎(下称慢乙肝)患者的血清及肝活检组织、99例乙型肝炎后肝硬化肝组织,分别用基因芯片、原位分子杂交法、免疫组织化学法、雅培试剂检测HBVDNA、HBcAg、HBsAg、HBeAg。结果15例慢乙肝HBsAg、HBeAg阳性患者的乙型肝炎血清基因芯片检测均阳性。15例肝组织标本中,免疫组化法HBcAg阳性15例,HBVDNA原位分子杂交阳性14例,基因芯片检测阳性14例。99例乙型肝炎后肝硬化肝组织标本中,HBcAg阳性67例、HBVDNA阳性53例,基因芯片检测阳性46例;32例HBcAg、HBVDNA阴性组织基因芯片检测均阴性。结论肝炎基因诊断芯片可以检测肝组织及血清HBVDNA,其诊断准确率高,假阳性率低。

s Objective [WT5”BZ]To study the application of the gene chips technology in detecting HBVDNA in the serums and liver tissues of patients with 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.HBVDNA,HBcAg,HBsAg and HBeAg in the serum samples and liver tissues of 15 patients with hepatitis B and HBVDNA, HBcAg in the liver tissues of the 99 patients with posthepatitic cirrhosis were detected and diagnosed by Abbott reagents,immunohistochemical method,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 positive diagnosed by the gene chips.15 cases expression of HBcAg and 14 cases expression of HBVDNA in liver tissues were positive detected respectively by immunohistochemical method,by in situ molecule hybridization technique,and by gene chip. In the 99 liver tissues samples of posthepatitic cirrhosis,67 cases expression of HBcAg and 53 cases expression of HBVDNA were positive detected with immunohistochemical method and in situ molecule hybridization technique, in which 46 cases expression of HBVDNA were positive detected by gene chip.32 cases of HBcAg and HBVDNA were negative detected by the three test methods.Conclusion The serums and liver tissues HBVDNA can be detected simultaneity by the gene chip,the accuracy of diagnosing is high and the rate of the sham positive is lower.