**4.20 Fluorescence melting curve analysis (FMCA)**

It is a probe-based assay that uses melting curve analysis to detect and differentiate between CPV-2 variants. This assay consists of 2 TaqMan probes namely FAM labeled and HEX labeled. The FAM-labeled probe sequence is perfectly complementary to CPV-2a, with a 1 bp mismatch to CPV-2b and a 2 bp mismatch to CPV-2c. The HEXlabeled probe has complete complementarity with the original CPV-2 and a 1-bp mismatch with the other variants. This method is also capable of detecting samples containing more than one variant without sequencing [72].


#### **Table 1.**

*Summary of the different types of diagnostic assays for CPVE diagnosis.*



**Table 2.**

*List of commericially available kits for CPV-2 detection.*
