Abstract
BackgroundPorcine parvovirus (PPV) is the important causative agent for infectious infertility, which is a fairly tough virus that multiplies normally in the intestine of pigs without causing clinical signs in the world.ResultsWe developed an assay integrating real-time PCR and high resolution melting (HRM) analysis for the detection of PPV. Primers targeting the VP gene were highly specific, as evidenced by the negative amplification of closely related viruses, such as porcine circovirus 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), classical swine fever virus (CSFV), or Japanese encephalitis virus (JEV). The performance of unlabeled real time PCR was compared to TaqMan real time PCR, and the detection limits of the two methods were nearly equal. Moreover, there was good correlation between Cp and diluted genomic DNA when tested with the two methods. The assay has the accuracy of 100% in reference to labeled real time PCR, when it was tested on 45 clinical samples.ConclusionsThe present study demonstrated that the established assay integrating real-time PCR and HRM is relatively cost-effective and more stable, which provides an alternative tool for rapid, simple, specific and sensitive detection of PPV.
Highlights
Porcine parvovirus (PPV) is the important causative agent for infectious infertility, which is a fairly tough virus that multiplies normally in the intestine of pigs without causing clinical signs in the world
We developed a real-time PCR assay coupled with high-resolution melting (HRM) analysis for the rapid and specific detection of PPV
Specificity of the detection assay and confirmation of amplicon identity The specificity of the primers was determined by performing PCR using pure genomic DNA from closely related viruses, including porcine circovirus 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), classical swine fever virus (CSFV), or Japanese encephalitis virus (JEV)
Summary
Porcine parvovirus (PPV) is the important causative agent for infectious infertility, which is a fairly tough virus that multiplies normally in the intestine of pigs without causing clinical signs in the world. Porcine parvovirus (PPV) is a major cause of reproductive failure in pigs. PPV does not cause clinical signs at any other time than during pregnancy, when it may cross the placenta and infect developing embryos and fetuses, resulting in resorption and mummification, abortion, stillbirth, neonatal death and reduced neonatal vitality [2]. Several methods have been developed for rapid detection of PPV, including specific PCR, ELISA and LAMP [4,5,6].
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