Brichacek B, Stevenson M
Program in Molecular Medicine, University of Massachusetts Medical Center, Worcester 01655, USA.
Methods. 1997 Aug;12(4):294-9. doi: 10.1006/meth.1997.0482.
Quantitative competitive PCR is a highly sensitive technique that allows accurate quantitation of small amounts of RNA. We have modified the original method to include the use of an internal standard at all stages of sample analysis. In this way, the method can accommodate for variations in the recovery of viral particles and in the isolation of genomic RNA as well as provide a suitable competitive substrate during quantitative RNA PCR. We have used this method to characterize changes in virus load in plasma of HIV-1-seropositive individuals following their vaccination against opportunistic infections.
定量竞争性聚合酶链反应是一种高度灵敏的技术，可对少量RNA进行精确定量。我们对原始方法进行了改进，在样本分析的所有阶段都使用内标。通过这种方式，该方法能够适应病毒颗粒回收和基因组RNA分离过程中的变化，并在定量RNA聚合酶链反应期间提供合适的竞争性底物。我们已使用此方法来表征HIV-1血清阳性个体在接种预防机会性感染疫苗后血浆中病毒载量的变化。
