title: Effects of DNA extraction method on Porcine circovirus-2 real-time polymerase chain reaction quantification in swine lymph node samples.
abstract: Quantitative real-time polymerase chain reaction (PCR) has become an important tool for Porcine circovirus-2 (PCV-2) research and diagnosis. However, significant differences in detection limit and quantification data, among laboratories and quantitative real-time PCR methods, have been demonstrated. New efforts are required for providing more accurate and comparable results. The current study is an evaluation of the effects of DNA extraction procedures on PCV-2 quantification in lymph node samples. Differences, greater than 1 log(10) copies/g, were shown among PCV-2 loads detected after different extraction procedures. The work highlighted the critical role of the DNA extraction method in PCV-2 quantification by quantitative real-time PCR. This important aspect should be evaluated when comparing data from different laboratories or different studies. The PCV-2 quantification data should not be considered comparable before demonstrating the equivalence of the DNA extraction methods performed.
authors.affiliation: IZSLER Diagnostic Section of Mantova, Circovallazione Sud 21/a, 46100 Mantova, Italy. silvia.faccini@izsler.it
authors.forename: Silvia
authors.initials: S
authors.lastname: Faccini
authors.forename: Arrigo D
authors.initials: AD
authors.lastname: Nigrelli
authors.forename: Giuliana
authors.initials: G
authors.lastname: Franzini
authors.forename: Carlo
authors.initials: C
authors.lastname: Rosignoli
authors.forename: Ilaria
authors.initials: I
authors.lastname: Barbieri
authors.forename: Giovanni L
authors.initials: GL
authors.lastname: Alborali
authors.forename: Maria B
authors.initials: MB
authors.lastname: Boniotti
mesh_terms: D000818:Animals; D017925:Circovirus / Q000145:classification* / Q000235:genetics*; D004279:DNA, Viral / Q000302:isolation & purification*; D008198:Lymph Nodes / Q000821:virology*; D053570:Porcine Postweaning Multisystemic Wasting Syndrome / Q000821:virology*; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary*; D013552:Swine; D019562:Viral Load
chemical_list: D004279:DNA, Viral