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Development of a Plexor real-time PCR assay for the detection of porcine circovirus type 2
Authors:Vlasakova Michaela  Jackova Anna  Leskova Valeria  Vilcek Stefan
Affiliation:University of Veterinary Medicine and Pharmacy, Komenskeho 73, 041 81 Kosice, Slovakia
Abstract:A novel, real-time PCR system for the detection of porcine circovirus type 2 (PCV2) was developed. The system employed Plexor technology and detected 108-101 copies per reaction of PCV2 DNA within a recombinant plasmid. The examination of clinical material showed consistent diagnostic sensitivity when samples contained more than 103 viral copies per reaction. Specificity of Plexor real-time PCR was confirmed using the porcine viruses PCV1, PRRSV, CSFV, TTSuV1 and TTSuV2 employing the melting curve analysis of PCR products. The low values of coefficient of variation in the intra- (1.74%) and inter-assay (2.41%) analysis suggested that the assay was a highly reproducible. The Plexor real-time PCR was compared with three other real-time PCR systems (SYBR Green, TaqMan, LUX) with conclusion that it can be used as a method of choice for the detection and quantitation of PCV2.
Keywords:Plexor   Real-time PCR   Porcine circovirus type 2   Detection   Quantitation
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