Development of Primer-Probe Energy Transfer real-time PCR for the detection and quantification of porcine circovirus type 2.
A real-time PCR assay, based on Primer-Probe Energy Transfer (PriProET), was developed to improve the detection and quantification of porcine circovirus type 2 (PVC2). PCV2 is recognised as the essential infectious agent in post-weaning multisystemic wasting syndrome (PMWS) and has been associated with other disease syndromes such as porcine dermatitis and nephropathy syndrome (PDNS) and porcine respiratory disease complex (PRDC). Since circoviruses commonly occur in the pig populations and there is a correlation between the severity of the disease and the viral load in the organs and blood, it is important not only to detect PCV2 but also to determine the quantitative aspects of viral load. The PriProET real-time PCR assay described in this study was tested on various virus strains and clinical forms of PMWS in order to investigate any correlation between the clinical signs and viral loads in different organs. The data obtained in this study correlate with those described earlier; namely, the viral load in 1 ml plasma and in 500 ng tissue DNA exceeds 10(7) copies in the case of PMWS. The results indicate that the new assay provides a specific, sensitive and robust tool for the improved detection and quantification of PCV2.
Bálint
Adám
A
Joint R&D Division, Departments of Virology, The National Veterinary Institute & The Swedish University of Agricultural Sciences, Ulls väg 2B, SE-751 89 Uppsala, Sweden. adambalint@yahoo.co.uk
Tenk
Miklós
M
Deim
Zoltán
Z
Rasmussen
Thomas Bruun
TB
Uttenthal
Ase
A
Cságola
Attila
A
Tuboly
Tamás
T
Farsang
Attila
A
Fossum
Caroline
C
Timmusk
Sirje
S
Berg
Mikael
M
Belák
Sándor
S
D000818:Animals; D001483:Base Sequence; D017925:Circovirus / Q000145:classification* / Q000302:isolation & purification*; D004279:DNA, Viral / Q000235:genetics; D004735:Energy Transfer; D016679:Genome, Viral; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D053570:Porcine Postweaning Multisystemic Wasting Syndrome / Q000821:virology; D012680:Sensitivity and Specificity; D013552:Swine
D004279:DNA, Viral