Development and Application of a qPCR Assay for Detection of Duck-Origin Goose Parvovirus
To develop a qPCR assay for rapid and accurate detection of Duck-origin goose parvovirus(D-GPV),we designed and synthesize specific primers based on the sequences of D-GPV gene.A recombinant plasmid was then constructed and verified.Subsequently,we used it as the positive template to draw the standard curve of this assay and then tested its sensitivity,specificity,repeatability and infectious samples.The results showed that the linear relationship of the standard curve of this method was good with the correlation coefficient at 0.999 and the sensitivity at 10 copies.Its specificity was also good as there was no cross-reactivity with other viruses.In the repeatability test,the inter-assay and intra-assay coefficients of variation were both lower than 1％with good stability.Examination of oropharyngeal and cloacal swabs from ducks infected with D-GPV showed that ducks shed viruses through mouths and cloacae at 1 dpi till 42 dpi.The viral loads reached the peak at 3 dpi.The qPCR method developed in this research had high sensitivity,specificity and stability,and provided a fast and accurate method for clinical detection of D-GPV.
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