Simultaneous detection and subtyping of porcine endogenous retroviruses proviral DNA using the dual priming oligonucleotide system.
The purpose of this study was to develop a multiplex PCR that can detect porcine endogenous retrovirus (PERV) proviral genes (pol, envA, envB, envC) and porcine mitochondrial DNA, using a dual priming oligonucleotide (DPO) system. The primer specifically detected the PERV proviral genes pol, envA, envB, envC, and porcine mitochondrial DNA only in samples of pig origin. The sensitivity of the primer was demonstrated by simultaneous amplification of all 5 target genes in as little as 10 pg of pig DNA containing PERV proviral genes and mitochondrial DNA. The multiplex PCR, when applied to field samples, simultaneously and successfully amplified PERV proviral genes from liver, blood and hair root samples. Thus, the multiplex PCR developed in the current study using DPO-based primers is a rapid, sensitive and specific assay for the detection and subtyping of PERV proviral genes.
Moon
Hyoung Joon
HJ
Department of Veterinary Medicine Virology Laboratory, College of Veterinary Medicine and BK21 Program for Veterinary Science, Seoul National University, Seoul 151-742, Korea.
Park
Seong Jun
SJ
Kim
Hye Kwon
HK
Ann
Soo Kyung
SK
Rho
Semi
S
Keum
Hyun Ok
HO
Park
Bong Kyun
BK
D000818:Animals; D017931:DNA Primers / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics*; D017988:Gammaretrovirus / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011533:Proviruses / Q000145:classification / Q000235:genetics*; D012680:Sensitivity and Specificity; D034421:Sus scrofa / Q000235:genetics* / Q000821:virology*
D017931:DNA Primers; D004272:DNA, Mitochondrial