南方医科大学学报 ›› 2016, Vol. 36 ›› Issue (11): 1496-.

• • 上一篇    下一篇

Allglo探针与TaqMan探针荧光定量RT-PCR检测猴免疫缺陷病毒的比较

吴胜男,谢延峥,刘翠华,何金洋   

  • 出版日期:2016-11-20 发布日期:2016-11-20

Comparison of TaqMan and Allglo probe in fluorescence quantitative RT-PCR detection of simian immunodeficiency virus

  • Online:2016-11-20 Published:2016-11-20

摘要: 目的比较Allglo 探针和TaqMan探针荧光定量RT-PCR检测SIV 的灵敏度下限和重复性。方法把SIV 标准品进行梯 度稀释成6 个浓度,每个浓度同时提取12 个标本进行批内差异分析,每个标本提取12 次进行批间差异分析之后进行逆转录 并用TaqMan探针和Allglo 探针进行定量PCR检测。批内差异分析者每个标本同时进行逆转录和上机检测,批间差异分析 者分12 次进行逆转录和检测,对这两种探针的PCR结果利用ABI7300 定量PCR仪所携带的软件和相关的统计学方法进行 分析。结果TaqMan 探针和Allglo 探针法检测SIV 标准品的灵敏度下限均为50 copies/mL。重复性结果显示:批内结果差 异分析显示Allglo 探针法最大变异系数为0.63%,最小变异系数为0.33%,TaqMan 探针法最大变异系数为1.33%,最小变异 系数为0.2%;批间结果差异分析显示Allglo 探针法最大变异系数为1.77%,最小变异系数为0.95%,TaqMan探针法最大变异 系数为1.86%,最小变异系数为1.03%。结论在荧光定量RT-PCR 法检测猴免疫缺陷病毒中,Allglo 探针法可能优于 TaqMan探针法。

Abstract: Objective To compare the sensitivity and reproducibility of Allglo and TaqMan probe in the detection of simian immunodeficiency virus (SIV) using fluorescence quantitative RT-PCR (QPCR). Methods The reference sample of SIV was diluted to 6 gradient concentrations; at each concentration 12 samples were tested to analyze the variations within batches, and each sample was tested for 12 times for analysis of variations between batches by QRT-PCR using TaqMan probe and Allglo probe. The results of QPCR using the two probes were analyzed with ABI7300 PCR system software. Results In QPCR using TaqMan and Allglo probe, the lower limit of sensitivity for SIV detection was both 50 copies/mL. Assessment of the reproducibility of the tests showed that the maximum and minimum coefficients of variation between batches were 0.63% and 0.33 % with Allglo probe, respectively, as compared with 1.33% and 0.2% with TaqMan probe. The maximum and minimum coefficients of inter-batch variation was 1.77% and 0.95% with Allglo probe, respectively, as compared with 1.86% and 1.03% with TaqMan probe. Conclusion Allglo probe shows a better performance then TaqMan probe in detection of SIV QPCR.