Grolla Allen
Special Pathogens Program, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, MB, Canada.
Methods Mol Biol. 2017;1628:341-352. doi: 10.1007/978-1-4939-7116-9_27.
Reverse transcriptase polymerase chain reaction (RT-PCR)-based techniques allow for highly sensitive and specific detection of RNA viruses. Detection of the amplification products can be achieved using several methods. The following are descriptions of the detection of ebolavirus RNA using end-point RT-PCR (agarose gel visualization of amplification products) and quantitative RT-PCR (Q-RT-PCR), with fluorescent detection using an intercalating dye or detection with the use of 5' hydrolysis probe assays. All of these techniques can be used to accurately detect the presence of ebolavirus in samples.
基于逆转录酶聚合酶链反应(RT-PCR)的技术能够对RNA病毒进行高度灵敏且特异的检测。扩增产物的检测可通过多种方法实现。以下是使用终点RT-PCR(扩增产物的琼脂糖凝胶可视化)和定量RT-PCR(Q-RT-PCR)检测埃博拉病毒RNA的描述,其中定量RT-PCR采用嵌入染料进行荧光检测或使用5'水解探针检测法。所有这些技术均可用于准确检测样本中埃博拉病毒的存在。