Objectives

 To develop multiplex PCR for identification of CPV and differentiation of CPV 2 from CPV 2a/2b.

 To develop a rapid, sensitivity and specificity tool for routinely diagnosis of CPV infected dogs.

Materials & Methods

Samples from various commercial CPV vaccine strains, both CPV2 and CPV2a/2b strain, and diarrheic dog's feces, clinically suspected as CPV infection, were performed DNA extraction. These extracts were then used as DNA templates. Two set of primers, CPV-NS1 (common primers for CPV) and CPV2a/2b-VP2 (strain specific primers), were independently tested for each PCR and, consequently, adjusted a condition for multiplex PCR. The results were visualized by agarose gel electrophoresis and UV illumination.

Results

NS1 primers gave the positive results for all vaccine strains and 10 of 12 fecal samples. However, VP2 primers gave the positive results for the vaccine using CPV2a or CPV2b and 10 positive fecal samples. Multiplex PCR using NS1 and VP2 primers also gave the similar results.

Conclusion

The multiplex PCR could be used to detect and differentiate CPV2 from CPV2a/2b from the feces of vaccinated and infected dogs.