Preclinical Detection of Porcine Circovirus Type 2 Infection Using an Ultrasensitive Nanoparticle DN

• 作 者: Yong Huang Xiujuan Zhang Qian Du Fengyu Wang Xiaomin Zhao Wenlong Zhang Dewen Tong
• 机 构: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100
• 出 处: 中国畜牧兽医学会兽医病理学分会第二十次学术研讨会暨中国病理生理学会动物病理生理专业委员会第十九次学术研讨会
• 发表日期: 2014年6期
• 关键词: PCV2 nanoparticle PCR

　　Porcine circovirus type 2 (PCV2) has emerged as one of the most important pathogens affecting swine production globally.Preclinical identification of PCV2 is very important for effective prophylaxis of PCV2-associated diseases.In this study,we developed an ultrasensitive nanoparticle DNA probe-based PCR assay (UNDP-PCR) for PCV2 detection.Magnetic microparticles coated with PCV2 specific DNA probes were used to enrich PCV2 DNA from samples,then gold nanoparticles coated with PCV2 specific oligonucleotides were added to form a sandwich nucleic acid-complex.After the complex was formed,the oligonucleotides were released and characterized by PCR.This assay exhibited about 500-fold more sensitive than conventional PCR,with a detection limit of 2 copies of purified PCV2 genomic DNAand 10 viral copies of PCV2 in serum.The assay has a wide detection range for all of PCV2 genotypes with reliable reproducibility.No cross-reactivity was observed from the samples of other related viruses including porcine circovirus type 1,porcine parvovirus,porcine pseudorabies virus,porcine reproductive and respiratory syndrome virus and classical swine fever virus.The positive detection rate of PCV2 specific UNDP-PCR in 40 preclinical field samples was 27.5%,which appeared greater than that by conventional and real-time PCR and appeared application potency in evaluation of the viral loads levels of preclinical infection samples.The UNDP-PCR assay reported here can reliably rule out false negative results from antibody-based assays,provide a nucleic acid extraction free,specific,ultrasensitive,economic and rapid diagnosis method for preclinical PCV2 infection in field,which may help prevent large-scale outbreaks.