A TaqMan qPCR for quantitation of Ungulate protoparvovirus 1 validated in several matrices

Journal of Virological Methods
André Felipe StreckUwe Truyen

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.

References

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Feb 14, 2006·Journal of Virological Methods·Sonja WilhelmUwe Truyen
Mar 16, 2006·Journal of Virology·Laura A Shackelton, Edward C Holmes
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Dec 2, 2008·Journal of Virological Methods·Hong-Ying ChenZhen-Ya Wang
Dec 3, 2008·Virus Research·Cui ShangjinJoaquim Segalés
Jul 29, 2011·The Journal of General Virology·André Felipe StreckUwe Truyen
May 12, 2012·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Dániel CadarAttila Cságola
Jun 28, 2013·The Journal of General Virology·André Felipe StreckUwe Truyen

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Citations

Dec 21, 2017·Viruses·István MészárosZoltán Zádori

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