Establishment of a TaqMan-based quantitative real-time PCR for the detection of porcine parvovirus.

Abstract

Porcine Parvovirus (PPV) is a non-enveloped DNA virus that predominantly induces reproductive disorders in swine. The ongoing emergence of novel PPV variants and the frequent co-infections with other viruses have led to significant economic losses within the swine industry. This study, utilizing 31 previously reported complete PPV genome sequences, identified a conserved fragment of the PPV-NS1 gene through homology analysis. A TaqMan-based real-time quantitative PCR (TaqMan-qPCR) detection method was developed targeting this specific fragment. Sensitivity assessments determined a detection limit of 8.5 copies/μL for standard plasmids. Specificity assessments showed no cross-reactivity with 10 other prevalent swine pathogens. The coefficients of variation for both intra-assay and inter-assay repeatability tests were both under 1%, demonstrating high reproducibility. Moreover, an analysis involving 32 clinical samples was conducted to compare the detection outcomes of the developed method with those obtained from a commercial kit. The findings demonstrated that the established method achieved a relative sensitivity of 100% and a relative compliance rate of 75%, suggesting its potential as an alternative to the commercial kit. In summary, the TaqMan-qPCR method developed in this study exhibits high sensitivity and specificity, making it ideal for detecting various clinical samples. It also provides a valuable tool for monitoring PPV and examining its epidemiological traits.