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Published on: December 3, 2011
A TaqMan qPCR for quantitation of Ungulate protoparvovirus 1 validated in several matrices
André Felipe Streck1, Francine Hergemöller2, Dana Rüster2
1Institute for Animal Hygiene and Veterinary Public Health, Faculty of Veterinary Medicine, University of Leipzig, An den Tierkliniken 1, 04103 Leipzig, Germany; Laboratory of Virology, Faculty of Veterinary Medicine, Rio Grande do Sul Federal University, Avenida Bento Gonçalves 9090, 91540-000 Porto Alegre, Brazil.
Abstract:
Ungulate protoparvovirus 1 (UPV1) is one of the major causes of reproductive disorders in swine. Recently, the rapid viral evolution of UPV1 and its viral persistence in several tissues has been described. Based upon this, a real-time qPCR method using upgraded primers targeting VP1 and applying the TaqMan technology was developed in this study for UPV1, and it was validated in feces, serum and tissue. Within the results, the limit of detection of the qPCR was 100copies of the viral genome per reaction of serum and feces and 1000copies of the viral genome per reaction of the grinded tissue (pre-inoculated matrices with diluted serially viruses). No cross reactivity was observed with other viruses associated with reproductive disorders. The assay was specific and reproducible, presenting low intra- and inter-assay variation (0.93% and 1.06%, respectively). In 50 clinical samples, the method was found to be more sensitive than immunofluorescence and a SYBR Green PCR. In conclusion, this qPCR represents an upgraded and useful tool to quantify UPV1 in different sample matrices for diagnostic and research purposes.
