Huttunen Moona, Mercer Jason
MRC-Laboratory for Molecular Cell Biology, University College London, London, UK.
Methods Mol Biol. 2019;2023:189-208. doi: 10.1007/978-1-4939-9593-6_12.
Quantitative PCR-based methods have proven to be easy-to-use, cost-effective procedures for the quantification of viral gene expression and viral genome numbers. Quantitative PCR (qPCR) and quantitative reverse transcriptase-PCR (qRT-PCR) are rapid and sensitive approaches that can be used to pinpoint defects in viral DNA replication and transcriptional activity, respectively. Due to the significant nucleotide overlap between Poxviridae these methods can be employed across a wide range of viruses from this family. Here we provide methods for the quantification of vaccinia DNA replication by qPCR and quantification of the three classes of vaccinia gene transcription by qRT-PCR.
基于定量PCR的方法已被证明是用于定量病毒基因表达和病毒基因组数量的易于使用且具有成本效益的程序。定量PCR（qPCR）和定量逆转录PCR（qRT-PCR）是快速且灵敏的方法，分别可用于查明病毒DNA复制和转录活性中的缺陷。由于痘病毒科之间存在显著的核苷酸重叠，这些方法可用于该科的多种病毒。在这里，我们提供了通过qPCR定量痘苗病毒DNA复制以及通过qRT-PCR定量三类痘苗病毒基因转录的方法。
