Comparative Efficacy of Conventional PCR and SYBR Green-based qPCR Assay for Detection of Canine parvovirus-2 in Diarrheic Pups from Nagpur

Background: Canine parvovirus (CPV) infection is an infectious and contagious viral disease of canines especially puppies of age under 6 months. CPV-2-infected dogs are characterized by a gastroenteritis disorder with clinical signs of anorexia, lethargy, vomiting, fever and diarrhoea. The present study was planned for the development of SY BR -Green-based real-time polymerase chain reaction for detection and quantitation of canine parvovirus type 2 in faecal samples of dogs and compare the efficacy with conventional PCR. Methods: A total of (n=200) samples from clinically ill dogs for parvovirus were tested by utilizing conventional PCR and qPCR based on SYBR green chemistry. A standard curve was plotted using a 10-fold serial dilution of standard DNA and Ct value. The standard curve was found linear over a 10-8 dilutions. The real-time PCR results were expressed as the number of DNA copies of CPV 2 per mg of faecal samples. Result: The results showed that out of 200 samples, 64 (32%) and 96(48%) samples were found positive by conventional PCR and SYBR green-based real-time PCR respectively. The qPCR showed range of 1.0x102 to 7.0x108. The present study suggested that qPCR was more sensitive and specific in the detection of CPV-2 as compared to conventional PCR.