ABSTRACT
Ungulate bocaparvoviruses (UBoV) 2-5 are recently discovered porcine bocaparvoviruses belonging to the family Parvoviridae, and are considered to be a potentially major cause of swine diseases. In order to detect local UBoV2 epidemics in China, we developed a TaqMan-based real-time PCR (qPCR) assay targeting the UBoV2 VP1 gene and compared the results of qPCR with conventional PCR (cPCR). The qPCR reproducibly detected a recombinant DNA plasmid containing the VP1 gene over a range of eight orders of magnitude, from 9.97 × 10-1-106 copies/µL, with a lower limit of detection of 9.97 copies/µL, compared with approximately 9.97 × 102 copies/µL for cPCR. The qPCR assay showed no cross-reactivity with other UBoVs or other porcine viruses. This qPCR assay detected UBoV2 in 18.1% (84/463) of pig samples collected from Chinese swine herds, with the highest infection rate of 35.3% (53/150) in loose stools. UBoV2 was not detected in liver samples. The TaqMan-based qPCR assay established in this study was highly sensitive and specific for the diagnosis and quantification of UBoV2. The results of this study will further our understanding of the etiology, epidemiology, and pathogenesis of UBoV2 infection.
Subject(s)
Bocavirus/isolation & purification , Parvoviridae Infections/veterinary , Real-Time Polymerase Chain Reaction/methods , Swine Diseases/diagnosis , Animals , Bocavirus/genetics , China , Feces/virology , Parvoviridae Infections/virology , Reproducibility of Results , Sensitivity and Specificity , Swine , Swine Diseases/virology , Viral Load/methodsABSTRACT
In this study, we obtained the whole genomes of three porcine bocaparvovirus (PBoV) strains (GD6, GD10, and GD23) by polymerase chain reaction. Sequence analysis showed that all three field strains belonged to PBoV group 3 (G3). The phylogenetic trees based on NS1, NP1, and VP1 differed to the extent that these PBoVs were potentially more closely related to bocaparvoviruses known to infect other animals than to other PBoVs. GD6, GD10, and GD23 all included the conserved sequences YLGPF and HDXXY, with known phospholipase A2 activity. Using recombination-detection software we identified a natural recombinant breakpoint in the NS1 region of PBoV G3. The results of this study will further the epidemiological characterization of PBoVs.
