[-Entrance of PCR for studying murine coronaviruses-]

Abstract

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.