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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HBVP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CV 1 5 %～ 2 0 % ,引物标记法用于检测HBV基因多态性灵敏度达 1 0 4copies 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
Objective To study the application of primer and incorporation markers in the detection of Hepatitis B virus (HBV) gene polymorphism. Methods Fluorescent molecule Cy5 was labeled with DNA or primers, then the HBVP and pre-C-C regions were amplified. The target fragment containing the fluorescent Cy5 was prepared and hybridized with the HBV gene polymorphism. The results were scanned and analyzed. Results The signal intensity of spiked labeling was slightly higher than that of primer, but the nonspecific signal intensity was also high. The detection of 42 sera from patients with hepatitis B by the two methods showed the same results and good reproducibility. The CV15% 20%, primer-labeled method for the detection of HBV gene polymorphism sensitivity of 104 copies ml. Conclusion Although the signal intensity of primer-labeled method is slightly lower than that of splicing method, the detection sensitivity and specificity still meet the clinical requirements