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
May 1, 1991·Journal of Virological Methods·T W MolitorL R Ludemann
Apr 16, 1999·Journal of Virological Methods·R M SoaresL J Richtzenhain
Jun 14, 2000·Animal Reproduction Science·W L MengelingA C Vorwald
May 29, 2003·The Journal of General Virology·Rodrigo Martins SoaresLeonardo José Richtzenhain
Sep 6, 2003·Journal of Immunological Methods·Joseph A WhelanMichael A Whelan
Jan 1, 2005·Proceedings of the National Academy of Sciences of the United States of America·Laura A ShackeltonEdward C Holmes
Dec 1, 2005·Journal of Veterinary Medicine. B, Infectious Diseases and Veterinary Public Health·S WilhelmU Truyen
Jan 25, 2006·The Journal of General Virology·P ZimmermannU Truyen
Feb 14, 2006·Journal of Virological Methods·Sonja WilhelmUwe Truyen
Mar 16, 2006·Journal of Virology·Laura A Shackelton, Edward C Holmes
Apr 25, 2007·Nucleic Acids Research·Warren A Kibbe
Dec 2, 2008·Journal of Virological Methods·Hong-Ying ChenZhen-Ya Wang
Dec 3, 2008·Virus Research·Cui ShangjinJoaquim Segalés
Feb 28, 2009·Clinical Chemistry·Stephen A BustinCarl T Wittwer
Dec 4, 2010·Virology Journal·Cuiping SongShangjin Cui
May 7, 2011·Molecular Biology and Evolution·Koichiro TamuraSudhir Kumar
Jul 29, 2011·The Journal of General Virology·André Felipe StreckUwe Truyen
Nov 29, 2011·Journal of Virological Methods·Lester J PérezHeidy Díaz de Arce
May 12, 2012·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Dániel CadarAttila Cságola
Jul 10, 2012·Journal of Virological Methods·Lan-lan ZhengBao-an Cui
Jan 23, 2013·Archives of Virology·André Felipe StreckUwe Truyen
Jun 28, 2013·The Journal of General Virology·André Felipe StreckUwe Truyen
Citations
Mar 5, 2016·The Journal of General Virology·Tessa FoersterUwe Truyen
Dec 21, 2017·Viruses·István MészárosZoltán Zádori
Mar 12, 2020·Veterinary Medicine·István KissVilmos Palya