引物标记与掺入标记在HBV基因多态性芯片检测中应用的研究

目的 研究引物标记与掺入标记在乙型肝炎病毒（HBV）基因多态性芯片检测中的应用。方法 用掺入标记或引物标记荧光分子Cy5，扩增HBV P区、前C／C区，制备含荧光分子Cy5的目的片段后，分别与HBV基因多态性芯片杂交、扫描并分析结果。结果 掺入标记法信号强度略高于引物标记法，但非特异信号强度也高，两法检测42份乙型肝炎患者血清，结果完全一致，重复性均较好，批内CV15％－20％，引物标记法用于检测HBV基因多态性灵敏度达10^4 copies/ml。结论 引物标记法信号强度虽略低于掺入标记法，但检测灵敏度、特异性仍满足临床要求。

Influence of labeled primer and labeled dUTP assays on the signal intensity of the chip for the detection of HBV gene polymorphism

Objective To evaluate the influence of assays with primer labeled with fluorochrome (Cy5) and dUTP labeled with Cy5 on the signal intensity of the chip for detection of hepatitis B virus (HBV) gene polymorphism.Methods The P region and pre C/C region of HBV gene were amplified by polymerase chain reaction (PCR) with Cy5 labeled primer or Cy5 labeled dUTP.The amplicons of the two assays were hybridized with chips,scanned and analyzed by computer software for the detection of HBV gene polymorphism.Results The signal intensity of assay with Cy5 labeled dUTP was slightly higher than that of assay with Cy5 labeled primer,but non specific signal intensity of the assay with Cy5 labeled dUTP was higher.The result of 42 samples showed that there was no significant difference between the two assays,and that both had a good repeatability and CV value (15% 20%).Conclusion The assay with Cy5 labeled primer may replace the assay with Cy5 labeled dUTP as a routine method to detect HBV gene polymorphism,and it is simpler and cheaper.