A real-time PCR method for detection of porcine circovirus 2

Abstract

To develop an accurate and rapid detection method for disease screening and prevention with porcine circovirus type 2 （PCV2）, a single-strand DNA virus that infects pigs, primers targeted to the ORF2 fragment of the PCV2 conserved region were designed. A real time polymerase chain reaction （PCR） method was developed using SYBR Green as a fluorescent dye and serial dilutions of ORF2 recombinant plasmid to construct a standard curve for an absolute quantification. Results showed that the detection limit was obtained with 101 copy DNA per microliter. This method exhibited 1 000 times higher sensitivity than conventional PCR, only specific identification of PCV2, and better repeatability with the less than 2% variation coefficient of intra- or inter-assay experiments. A total of 34 PCV2 positive clinical samples were confirmed using this real time PCR method, demonstrating 100% agreement in comparison with the conventional PCR having only 50% agreement. This real time PCR method could provide a valuable tool for PCV2 prevention and control.［Ch, 4 fig. 4 tab. 26 ref.］