1.Development Of Two Assays For Detection Of Wild-type Pseudorabies Virus 2.Screening Of Long Noncoding RNAs That Regulate The Replication Of Classical Swine Fever Virus

Pseudorabies?PR?,also known as Aujeszky's disease,is an economically important infectious disease of pigs and other animals caused by pseudorabies virus?PRV?.Since late 2011,emerging PRV variants have been confirmed to be responsible for the outbreaks.Here,we developed a triplex real-time PCR for differential detection of classical,variant and Bartha-K61 vaccine strains of PRV.The sensitivity was determined to be 50,50 and 5 copies for the TJ,SC and Bartha-K61 strain,respectively.When testing a total of 234 clinical swine samples,the agreement between the triplex real-time PCR and virus isolation was 100%?234/234?for classical strains,99.5%?233/234?for variant strains,and 100%?234/234?for the Bartha-K61 vaccine strain.The results demonstrate that this method is sensitive and specific and will be useful for differentiation of diverse PRV strains.This study also presents a novel assay based on cross-priming amplification in combination with gold nanoparticle lateral flow strip?CPA-strip?for rapid and on-site detection of wild-type PRV.In the assay,FITC and biotin were labeled at the 5'-ends of the two probes,respectively,resulting in amplified products labeled with either FITC or biotin at the 5'-end.Then the amplified products?biotin-dsDNA-FITC?,as novel targets,were recognized by standardized strips.The detection limit was determined to be 200 copies.When testing a total of 293 field swine samples,the agreement between the CPA-strip assay and a triplex real-time PCR or virus isolation were 100%?293/293?or 97.3%?285/293?.Collectively,the assay is specific,sensitive and convenient that enables rapid and on-site diagnosis of PRV infections.Long noncoding RNAs?IncRNAs?are involved in the host antiviral responses through regulating the expression of adjacent immune genes.Based on RNA sequencing technology,we found more than 4,000 novel IncRNAs from CSFV-infected peripheral blood mononuclear cells.Six IncRNAs are located proximally to the antiviral genes and the expression of the IncRNAs and adjacent genes shows a positive correlation.Therefore,we hypothesize that the expression of these antiviral genes may be regulated by IncRNAs.The result shows that lncRNA₂522 can inhibit the expression of OAS2.