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

发布时间：2018-06-18 23:07

本文选题：Allglo探针 + TaqMan探针　；参考：《南方医科大学学报》2016年11期

【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/m L。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 lower limit and repeatability of Allglo probe and TaqMan probe fluorescence quantitative RT-PCR for the detection of SIV. Methods SIV standard samples were diluted into 6 concentrations, each concentration was extracted at the same time, 12 specimens were extracted at the same time for intra batch difference analysis. Each specimen was extracted 12 times after batch difference analysis and used TaqMan probe. The Allglo probe was used for quantitative PCR detection. Each sample in the group was simultaneously reverse transcriptase and on machine detection, and the inter batch difference analyst was divided into 12 times for reverse transcription and detection. The PCR results of the two probes were analyzed by the software and related statistical methods carried by the ABI7300 quantitative PCR instrument. Results TaqMan probe and Allglo The sensitivity lower limit of SIV standard samples by probe method was 50 copies/m L. repeatability results showed that the maximum variation coefficient of Allglo probe method was 0.63%, the minimum variation coefficient was 0.33%, the maximum variation coefficient of TaqMan probe method was 1.33% and the minimum variation coefficient was 0.2%. The difference analysis between batch results showed Allglo probe method The maximum coefficient of variation is 1.77%, the minimum variation coefficient is 0.95%, the maximum variation coefficient of TaqMan probe method is 1.86% and the minimum variation coefficient is 1.03%.. The Allglo probe method may be superior to the TaqMan probe method in the detection of monkey immunodeficiency virus by fluorescence quantitative RT-PCR method.
【作者单位】：广州中医药大学热带医学研究所;
【基金】：国家自然科学基金(81573861) 广州中医药大学青年英才基金(A3-AAC414124A03)共同资助~~
【分类号】：R440

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