Hepatitis gene chip in detecting HBV DNA, HCV RNA in serum and liver tissue samples of hepatitis patients

摘要

OBJECTIVE: To study the preparation of diagnostic gene chip for detecting hepatitis B virus (HBV) andhepatitis C virus (HCV) and its accuracy in detecting HBV DNA and HCV RNA in serum and liver tissues.METHODS: The probes, which depend on the conservative gene fragment of hepatitis virus, wasdesigned, synthesized and spotted on the modified glass. The probes and some other control probes wereassembled on the diagnostic microarray of hepatitis virus. The gene of hepatitis virus, purified from bloodor tissue, was labeled with fluorescence and hybridized to the microarray. The hybridized microarry wasscanned with microarray scanner and the diagnostic result was analyzed from the scanning data. Fourtypatients with hepatitis B virus and 40 healthy people or 40 patients with hepatitis C virus were subjected todetection of HBV DNA and HCV RNA with the hepatitis virus gene chip by the double-blind method.Paraffin liver specimens obtained from 99 cases of posthepatitic cirrhosis were used to detect HBV DNA.The liver tissues and serum from 15 cases of chronic hepatitis B were used to detect HBV DNA.Simultaneously, HBsAg and HBcAg were detected in the serum by fluorescence microparticlequantitation, HBV DNA and HCV RNA in the serum by PCR, and HBcAg in liver tissues byimmunocytochemistry or HBV DNA by in situ molecular hybridization.RESULTS: Chip detection of serum specimens showed that 30 patients were HBV DNA positive and 10HBV DNA negative in the 40 patients with HBV positive, 25 patients were HCV RNA positive and 15patients were HCV RNA negative in the 40 patients with HCV positive, and all were HBV and HCVnegative in the 40 healthy people. In 15 patients with HBV marker positive who were subjected to liverbiopsy, 15 patients were detected HBV DNA positive in serum by gene chip, 15 patients HBcAg positivein liver tissues by immunocytochemistry, 14 patients HBV DNA positive in liver tissues by in situmolecular hybridization, and 14 patients HBV DNA positive in liver tissues by gene chip. Paraffin livertissues specimens from the 99 patients with posthepatitis B cirrhosis showed that 67 patients were detectedHBcAg positive by immunocytochemistry, 53 patients HBV DNA positive by in situ molecularhybridization, and 46 patients HBV DNA positive by gene chip. In the 46 patients, 40 patients weredetected HBV DNA and HBcAg positive by in situ molecular hybridization and immunocytochemistry, 6patients only HBcAg positive, and 33 patients HBcAg negative.CONCLUSIONS: The designed diagnostic gene chip can be used to simultaneously detect serum HBVDNA and HCV RNA, but the positive rate of HCV RNA diagnosed by this chip is lower. The gene chipcan detect HBV DNA in serum and in liver tissue.