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目的 建立SYBR Green I Real-time PCR检测HBV-DNA方法，并检测IFN-CSP对HepG2.2.15细胞内HBV-DNA影响。方法 提取HepG2.2.15细胞DNA，PCR扩增后纯化，PCR纯化产物梯度稀释作为标准品，采用SYBR Green I Real-time PCR 检测，建立标准曲线，并分析方法的特异性、灵敏度、重复性和稳定性。运用建立的SYBR Green I 方法检测IFN-CSP对HepG2.2.15细胞内HBV-DNA影响，并与Taqman方法进行比较。结果 SYBR Green I Real-time PCR方法特异性、重复性及稳定性均较好，线性范围为107~102拷贝，检测灵敏度可达102拷贝。IFN-CSP对HepG2.2.15细胞内HBV-DNA具有抑制效果，且呈剂量依赖性。SYBR Green I方法与Taqman商业试剂盒检测结果差异无统计学意义。结论 SYBR Green I Real-time PCR方法简便快速、结果准确、价格低廉，可以满足HBV-DNA拷贝数检测的需要。
OBJECTIVE: To establish a real-time PCR method for detection of HBV-DNA by SYBR Green I and to detect the effect of IFN-CSP on HBV-DNA in HepG2.2.15 cells. Methods The DNA of HepG2.2.15 cells was extracted and purified by PCR amplification. The products were purified by gradient dilution of PCR products. The standard curve was established by SYBR Green I Real-time PCR. The specificity, sensitivity, reproducibility and stability Sex. The established SYBR Green I method was used to detect the effect of IFN-CSP on HBV-DNA in HepG2.2.15 cells and compared with Taqman method. Results The SYBR Green I Real-time PCR method showed good specificity, repeatability and stability with a linear range of 107-102 copies and a detection sensitivity of 102 copies. IFN-CSP had inhibitory effect on HBV-DNA in HepG2.2.15 cells in a dose-dependent manner. There was no significant difference between SYBR Green I method and Taqman commercial kit. Conclusion The method of SYBR Green I Real-time PCR is simple, rapid, accurate and inexpensive and can meet the needs of HBV-DNA copy number detection.