磁珠法测定痕量肝炎病毒的研究

Study on trace hepatitis virus detected by magnetic beads

目的寻求灵敏而简便的肝炎病毒临床检测新方法。方法采用氧化硅包覆的磁珠同时提取肝炎病毒的DNA及RNA,结合荧光实时定量聚合酶链反应(PCR)检测乙型肝炎病毒(HBV)及丙型肝炎病毒(HCV)。结果磁珠法可以同时提取HBV-DNA和HCV RNA,磁珠法提取HBV-DNA的效率高于离心柱法和煮沸法,磁珠法提取HCV RNA的效率与离心柱法相近,但提取效率都高于煮沸法。结论将磁珠的富集分离与高灵敏度的检测手段相结合,可显著提高检测的特异性和灵敏度,且方法简便可自动化操作,可以满足大批样品肝炎病毒筛选检测的要求,适用于血液中心筛查及医院检测等领域。

Objective To find a new sensitive and simple clinical testing method to detect hepatitis virus. Methods The silica-coated magnetic beads were adopted to extract hepatitis virus DNA and RNA simultaneously, and combined with the real time PCR method for detecting hepatitis B virus and hepatitis C virus. Results The mag- netic beads could extract hepatitis B virus（HBV） DNA and hepatitis C virus（HCV） RNA simultaneously. The ex- traction efficiency of HBV-DNA by the magnetic beads method was higher than that by the centrifuge method and the boiling method,which of HCV RNA was close to the centrifuge method,but higher than the boiling method. Conclu- sion To combine the magnetic beads enrichment and separation of low intensity of virus with the high sensitivity de- tection method can significantly improve the detection specificity and sensitivity. The method is simple and can be au- tomated,which can meet the requirement of hepatitis virus screening detection in large number of samples, therefore, is suitable for the fields of blood center screening and hospital detection.