›› 2009, Vol. 21 ›› Issue (3): 0-224.

• 论文 •    

Development of SYBR Green I real-lime PCR assay for detection of porcine parvovirus

LI Ming-feng;W EI Zhan-yong;WANG Xue—bin;ZHANG Hong-ying;WANG Ya-bin;CUl Bao-an;*   

  1. 1 College of Animal Science and Veterinary Medicine,Henan argricultural University,Zhengzhou,Henan 450002,China;2 Henan Key Laboratory for Animal Food Safety,Zhengzhou,Henan 450002,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-05-25 Published:2009-05-25

Abstract: A 194 bp region of the porcine parvovirus VP2 gene was amplified by PCR and cloned into pGEM T vector,named as pGEM-VP2.Serial dilutions of plasmid pGEM-VP2 were used as standard templates for PCR to quantify the virus
genomic copy number.We develoloped a SYBR Green I real-time PCR to detect porcine parvovirus.Sensitivity analysis showedthat the developed SYBR GreenI real-time PCR could detect 1.0×10 2 template/μL.The speeitleity assay exhibited that negative control and the other porcine pathogens,such as PRRSV,PCV,JEV,PRV,HCV and SIV could not be detected by this PCR.It was showed that10 positive results could be observed by the real-time PCR for 10 suspicious positive samples,but 7 positive results by normal PCR.The results suggested that as a result of the good sensitivity and specificity of the assay with a rapid and simple procedure,the real-time PCR method would be useful for the clinical diagnosis of porcine parvovirus infection.

Key words: porcine parvovirus, real-time PCR, standard curve