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Application of transcription mediated amplification and real-time reverse transcription polymerase chain reaction in detection of human immunodeficiency virus RNA



Application of transcription mediated amplification and real-time reverse transcription polymerase chain reaction in detection of human immunodeficiency virus RNA



Zhong Nan Da Xue Xue Bao. Yi Xue Ban 42(7): 776-782



目的:利用转录介导的扩增技术(transcription mediated amplification,TMA) 扩增HIV RNA,探讨TMA 技术扩增HIV RNA的敏感性及其与实时荧光定量反转录聚合酶链反应的比较。方法:利用TaqMan探针、特异性引物、鼠白血病反转录酶、T7-RNA聚合酶及PCR底物等建立TMA 扩增体系。通过扩增一组10倍梯度稀释的HIV RNA转录标准品,评价TMA 体系的灵敏性。收集60例HIV感染患者的血浆,同时采用TMA试剂与Cobas Amplicor HIV-1 Monitor 1.5版试剂进行检测,比较两种方法的阳性检出率,并利用线性回归和Bland-Altman法分析两种技术的相关性和一致性。结果:成功建立了TMA 扩增体系,这种技术可以检测低至10 copies/mL 的HIV转录标准品。60份HIV感染者血浆样本中,TMA及Cobas均检测到阳性46 份,均阴性12份,TMA检测阴性而Cobas 检测阳性样本2份,检测一致率为96.7%。两种技术阳性检出率差异无统计学意义(P>0.05)。对其中46 份TMA 检测和Cobas检测均有定量结果的血浆进行线性回归分析,两种技术有非常好的相关性(r=0.997,P<0.001)。Bland-Altman分析显示两种检测方法定量Lg差值平均值为0.02,44份(95.7%)样本在95%的一致性界限内。结论:TMA 技术具有高灵敏性潜能。TMA 与real-time RT-PCR 检测血浆中HIV RNA具有非常好的相关性与一致性。.

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Accession: 059389892

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PMID: 28845000


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