A sensitive method to detect canine parvoviral DNA in faecal samples by nested polymerase chain reaction

Abstract

Canine parvovirus 2 (CPV-2) is the causative agent of acute hemorrhagic enteritis and myocarditis in dogs. A nested polymerase chain reaction (n-PCR) was developed using published pCPV-2ab as external primer set and self designed and custom synthesized pCPV-2N as internal primer set for the detection of canine parvovirus in faecal samples of dogs. Out of 52 faecal samples, 27 and 31 were tested positive by one step PCR and n-PCR, respectively. Among the different breed of dogs, German shepherd was found to be highly susceptible (69.2%), followed by non-descript breeds (63.1%) and overall positivity was 59.6%. Ten-fold dilution of known cell culture supernatants was tested and it was observed that one step PCR had a detection limit of 10^–5 dilution, whereas nested-PCR had up to 10^–8 dilution. Thus, the nested-PCR seems to be a sensitive, specific and practical method for the detection of CPV in faecal samples.