title: A nanoparticle-assisted PCR assay to improve the sensitivity for rapid detection and differentiation of wild-type pseudorabies virus and gene-deleted vaccine strains.
abstract: Nanoparticle-assisted polymerase chain reaction (nanoPCR) is a novel method for the rapid amplification of DNA and has been adopted for the detection of virus because of its simplicity, rapidity, and specificity. A nanoPCR assay was developed to detect and differentiate wild-type and gene-deleted pseudorabies virus (PRV). Three pairs of primers for nanoPCR developed in this study were selected from conserved regions of PRV, producing specific amplicons of 431 bp (gB), 316 bp (gE), and 202 bp (gG). The sensitivity of this assay using purified plasmid constructs containing the specific gene fragments was 100-1000 fold higher than conventional PCR. The PRV nanoPCR assay did not amplify porcine parvovirus, porcine circovirus type 2, porcine reproductive and respiratory syndrome virus, porcine teschovirus, or African swine fever virus but produced three bands of expected size with PRV and two bands of expected size with the gene-deleted PRV-Bartha-K61. Of 110 clinical samples collected from seven provinces in China, 53% and 48% were positive for wild-type PRV according to the nanoPCR assay and virus isolation, respectively.
authors.affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, China.
authors.forename: Xingjie
authors.initials: X
authors.lastname: Ma
authors.forename: Yuchao
authors.initials: Y
authors.lastname: Cui
authors.forename: Zheng
authors.initials: Z
authors.lastname: Qiu
authors.forename: Bingkun
authors.initials: B
authors.lastname: Zhang
authors.forename: Shangjin
authors.initials: S
authors.lastname: Cui
mesh_terms: D000818:Animals; D002681:China; D017931:DNA Primers / Q000235:genetics; D017353:Gene Deletion; D011558:Herpesvirus 1, Suid / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D053758:Nanoparticles; D016133:Polymerase Chain Reaction / Q000379:methods*; D022421:Pseudorabies Vaccines / Q000235:genetics*; D012680:Sensitivity and Specificity; D013552:Swine; D014759:Viral Envelope Proteins / Q000235:genetics; D014773:Virology / Q000379:methods*
chemical_list: D017931:DNA Primers; D022421:Pseudorabies Vaccines; D014759:Viral Envelope Proteins; C089580:glycoprotein E, Suid herpesvirus 1; C070915:glycoprotein gII, pseudorabies virus
keywords: NanoPCR; PRV; gB; gE; gG