Homberger F R, Zhang L, Thomann P E
Institut für Labortierkunde, Veterinärmedizinische Fakultät, Universität Zürich.
Schweiz Arch Tierheilkd. 1996;138(2):67-73.
This paper describes a number of applications of the Polymerase Chain Reaction (PCR) in the study of enterotropic murine coronaviruses [mouse hepatitis virus (MHV)]. A diagnostic PCR was developed which detected all of 11 different MHV strains. This fast and reliable method was also able to differentiate MHV from other non-murine coronaviruses. On the basis of this assay a quantitative PCR was designed using a mutant template containing a point mutation which competed for the PCR primers. The amplification and cloning of the structural protein genes of enterotropic MHV strains it plasmid vectors for subsequent sequencing is described. In addition an RT PCR was developed which was able to selectivity detect artificially generated recombinant coronavirus.
本文描述了聚合酶链反应(PCR)在嗜肠性鼠冠状病毒[小鼠肝炎病毒(MHV)]研究中的一些应用。开发了一种诊断性PCR,可检测11种不同的MHV毒株。这种快速可靠的方法还能够区分MHV与其他非鼠冠状病毒。基于该检测方法,使用含有一个点突变的突变模板设计了一种定量PCR,该模板可与PCR引物竞争。文中描述了嗜肠性MHV毒株结构蛋白基因在质粒载体中的扩增和克隆,以便后续测序。此外,还开发了一种RT-PCR,它能够选择性地检测人工构建的重组冠状病毒。