Abstract: Partial region of the PRRSV GP5 gene was amplified by RT-PCR and cloned into pGEM-T Easy vector. Serial dilutions of the recombinant plasmid were used as standard templates for RT-PCR to quantify the virus genomic copy number. A SYBR GreenⅠreal\|time PCR was developed to detect PRRSV. Sensitivity analysis showed that the developed SYBR Green I real-time PCR could detect 10 copy/μL. The specificity assay exhibited that the other porcine pathogens, such as PPV, PCV, JEV, PRV, HCV and SIV could not be detected by this method. Then the established method was used to detect the clinical samples. The results showed that 15 positive samples out of 16 suspicious positive samples could be observed by real-time PCR and 12 positive samples could be detected by normal RT-PCR. All the results suggested that the real-time PCR we established in current study showed the characteristics of sensitivity and specificity, and could be used in clinical diagnosis and epidemiological investigation .

Key words: PRRSV, GP5 gene, real-time PCR, SYBR GreenⅠ