凝胶图像处理系统定量检测乙型肝炎病毒DNA

目的建立一种聚合酶链反应(PCR)-凝胶图像处理系统定量分析核酸扩增产物的方法。方法1.用PCR荧光定量分析仪实时监测PCR扩增曲线，并选择合适的扩增循环次数；2.将梯度标准乙型肝炎病毒(HBV)阳性血清(101～106拷贝/μl)分别进行40，35及32次循环次数的扩增，经琼脂糖电泳进行凝胶分析；3.选择凝胶分析软件中能客观反映核酸实际含量的最适参数，制定标准曲线。结果40及35次循环时，在101～104拷贝/μl线性良好，而在104，105及106拷贝/μl3个梯度时直线上升缓慢，趋于平坦。32次循环时，10拷贝/μl未能检出，102～106拷贝/μl的5个梯度的模板(对数值)与产物(体积)有良好的线性关系(r=0.97)。结论32次循环线性范围较广，为最适循环次数；在UVIband的吸光度定量分析指标中，以体积或体积百分比作为定量指标较好。

Quantitative detection of HBV-DNA on gel documentation systems

Objective To establish polymerase chain reaction-gel documentation systems (PCR-GDS) technology for detecting nucleic acid amplification products. Methods 1.PCR amplification curve was observed with fluorescence quantitative PCR (FQ-PCR), and suitable cycle numbers were selected. 2.Standard HBV-DNA sera (101-106 copies/μl) were amplified 40, 35 and 30 cycles separately. The PCR products were gel electrophoried and EB stained, then analyzed with GDS. Results When cycles were 40 and 35, linearity was good in standard sera 101-104 copies/μl, but line was nearly flat in 104-106 copies/μl; when cycles were 32, 10 copies/μl can not be detected, and linearity was good in 102-106 copies/μl (r=0.98). Conclusion 32 cycles were suitable cycle number for PCR-GDS. Among quantitative analysis of GDS, parameters of volume and volume percentage were better than others.