本文選題：Allglo探針 + TaqMan探針　； 參考：《南方醫(yī)科大學學報》2016年11期

【摘要】：目的比較Allglo探針和TaqMan探針熒光定量RT-PCR檢測SIV的靈敏度下限和重復性。方法把SIV標準品進行梯度稀釋成6個濃度,每個濃度同時提取12個標本進行批內差異分析,每個標本提取12次進行批間差異分析之后進行逆轉錄并用TaqMan探針和Allglo探針進行定量PCR檢測。批內差異分析者每個標本同時進行逆轉錄和上機檢測,批間差異分析者分12次進行逆轉錄和檢測,對這兩種探針的PCR結果利用ABI7300定量PCR儀所攜帶的軟件和相關的統(tǒng)計學方法進行分析。結果 TaqMan探針和Allglo探針法檢測SIV標準品的靈敏度下限均為50 copies/m L。重復性結果顯示:批內結果差異分析顯示Allglo探針法最大變異系數(shù)為0.63%,最小變異系數(shù)為0.33%,TaqMan探針法最大變異系數(shù)為1.33%,最小變異系數(shù)為0.2%;批間結果差異分析顯示Allglo探針法最大變異系數(shù)為1.77%,最小變異系數(shù)為0.95%,TaqMan探針法最大變異系數(shù)為1.86%,最小變異系數(shù)為1.03%。結論在熒光定量RT-PCR法檢測猴免疫缺陷病毒中,Allglo探針法可能優(yōu)于TaqMan探針法。
[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.
【作者單位】： 廣州中醫(yī)藥大學熱帶醫(yī)學研究所;
【基金】：國家自然科學基金(81573861) 廣州中醫(yī)藥大學青年英才基金(A3-AAC414124A03)共同資助~~
【分類號】：R440

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