Optimization The Procedure To Detect Porcine Circovirus 3 (Pcv3) Using Direct Pcr From Whole Blood

Abstract

A new swine pathogen that is becoming more and more significant to the global economy is Porcine circovirus 3 (PCV3). It has been discovered in the tissues of pigs around the world suffering from reproductive failure, myocarditis, multisystemic inflammation, and porcine dermatitis and nephropathy syndrome (PDNS). The diagnosis of porcine circovirus-associated disease (PCVAD) involves a combination of clinical symptoms, microscopic and gross lesions, and viral detection. Several PCR assays for PCV3 have been developed using tissues, serum, and semen as templates. However, current studies highlight limitations in cross-contamination, speed, cost-effectiveness, resource conservation, and field application. As such, tests that are rapid, reliable, and easily available are desperately needed for regular diagnosis as well as research purposes. It is thus imperative to shorten sample preparation procedure as well as choose a detection template that is easily collected. A direct PCR assay will eliminate the need for DNA extraction that so many PCR tests required. Among tissues from pig that PCV3 targeted, blood is readily accessible and carries a substantial amount of virus. The purpose of this study is to describe the optimization of a direct PCR assay from whole blood that targets the conserved cap gene to detect PCV3. For comparison, the optimization of a traditional PCR assay was done concurrently with the direct assay optimization. The results demonstrated excellent sensitivity, specificity, and efficiency. The optimized direct PCR assay, which eliminates the requirement for DNA extraction, provides a detection methodology that is inexpensive in cost, quick in processing time, has low cross contamination, and is widely applicable.