**4.18 Insulated isothermal PCR method**

It is a convection-based method using a hydrolysis probe for detection of CPV-2 and its antigenic variants. The reaction mixture is sequentially allowed to pass in an automatic manner through variable temperature zones in a capillary tube which undergoes thermocyclic phase to amplify the DNA and the probe hydrolysis produces optical output providing the result within an hour [70].
