A SimpleProbe(®) real-time PCR assay for differentiating the canine parvovirus type 2 genotype

Hoang, Minh; Wu, Hung-Yi; Lien, Ying-Xiu; Chiou, Ming-Tang; Lin, Chao-Nan

All genotyping results from the SimpleProbe ® assay were identical to the DNA sequencing results (Table S1 ). The specificity of this SimpleProbe ® assay was 100% in all CPV-2 genotypes. We also evaluated ten samples that could not be evaluated by DNA sequencing due to low viral loads (cq>35). The SimpleProbe ® assay identified one samples as genotype CPV-2c, and the remaining nine samples were negative (data not shown). (Figure 3) . Both The SimpleProbe ® assay required less time for genotyping than the conventional sequencing method. Moreover, the large sample size could be tested simultaneously, and the SimpleProbe ® assay was cheaper than DNA sequencing. Overall, the SimpleProbe ® assay offers some distinct advantages for genotyping CPV-2: (a) it reduces the cost and time required for DNA sequencing; (b) data are easy to interpret; (c) only a single-labeled sensor probe is used; and (d) it offers excellent specificity.