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目的建立一种简捷、实用的荧光定量PCR试剂质量评估方法。方法以扩增反应后的荧光背景、杂光及其试剂盒标准曲线的斜率与截距等多参数对其实施质量监控。结果不同品牌荧光定量PCR试剂盒标记探针的荧光强度以及背景荧光差异较大,以试剂盒a较为理想,其标准曲线斜率和截距的批内变异分别为3.106%和1.714%,试剂盒的批间变异为1.229%和2.14%。结论利用扩增反应后的荧光背景、杂光及其试剂盒标准曲线斜率与截距对试剂盒实施动态质量监控,较经典的血清盘测定法更为便捷、实用。
Objective To establish a simple and practical method for the quality assessment of fluorescent quantitative PCR reagents. Methods The quality control of the fluorescent background, stray light and the standard curve of the kit, such as slope and intercept, was carried out. Results The fluorescent intensity and background fluorescence of the labeled probes of different brands of fluorescent quantitative PCR kits were quite different. The kit a was ideal with the intra-assay variation of the slope and intercept of the standard curve of 3.106% and 1.714%, respectively. Inter-batch variation was 1.229% and 2.14%. Conclusion The dynamic quality control of the kit is achieved by using the fluorescence background and stray light of the amplification reaction and the slope and intercept of its standard curve of the kit, which is more convenient and practical than the classical serum plate assay.