Abstract
A TaqMan-based real-time polymerase chain reaction (PCR) assay was devised for the detection of porcine parvovirus (PPV). Two primers and a TaqMan probe for the non-structural protein NS1 gene were designed. The detection limit was 1 × 102 DNA copies/μL, and the assay was linear in the range of 1 × 102 to 1 × 109 copies/μL. There was no cross-reaction with 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 assay was specific and reproducible. In 41 clinical samples, PPV was detected in 32 samples with the real-time PCR assay and in only 11 samples with a conventional PCR assay. The real-time assay using the TaqMan-system can therefore be practically used for studying the epidemiology and management of PPV.
Highlights
Porcine parvovirus (PPV) is an autonomous parvovirus belonging to the genus parvovirus, subfamily Parvovirinae, family Parvoviridae
PPV is composed of a linear single-stranded segment of DNA approximately 5 kb long (Molitor, T.W., 1983), and its genome has more than two open reading frames (ORF) [6]
The conventional polymerase chain reaction (PCR) assay showed a negative result when the solution was diluted to 1.00 × 104 copies/μL. These results indicate that, based on direct observation, the sensitivity of the real-time PCR is 100 times greater than that of the conventional PCR (Figure 1C)
Summary
Porcine parvovirus (PPV) is an autonomous parvovirus belonging to the genus parvovirus, subfamily Parvovirinae, family Parvoviridae. It is one of the major etiological agents of reproductive failure in pigs. Reproductive failure caused by PPV is characterized by embryonic and foetal death, mummification, stillbirth, and delayed return to oestrus [1]. PPV has been implicated as the causative agent of diarrhea, skin disease, and arthritis in swine [2]. PPV has been reported from many different countries [3,4,5]. PPV is composed of a linear single-stranded segment of DNA approximately 5 kb long (Molitor, T.W., 1983), and its genome has more than two open reading frames (ORF) [6]. The 3’ end of ORF1 encodes nonstructural proteins (NS proteins), and the 5’ end of ORF2 encodes structural proteins (VP proteins)
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