title: Development of a nanoparticle-assisted PCR (nanoPCR) assay for detection of mink enteritis virus (MEV) and genetic characterization of the NS1 gene in four Chinese MEV strains.
abstract: BACKGROUND
  Mink enteritis virus (MEV) causes mink viral enteritis, an acute and highly contagious disease whose symptoms include violent diarrhea, and which is characterized by high morbidity and mortality. Nanoparticle-assisted polymerase chain reaction (nanoPCR) is a recently developed technique for the rapid detection of bacterial and viral DNA. Here we describe a novel nanoPCR assay for the clinical detection and epidemiological characterization of MEV.
  
  
  RESULTS
  This assay is based upon primers specific for the conserved region of the MEV NS1 gene, which encodes nonstructural protein 1. Under optimized conditions, the MEV nanoPCR assay had a detection limit of 8.75 × 10(1) copies recombinant plasmids per reaction, compared with 8.75 × 10(3) copies for conventional PCR analysis. Moreover, of 246 clinical mink samples collected from five provinces in North-Eastern China, 50.8% were scored MEV positive by our nanoPCR assay, compared with 32.5% for conventional PCR. Furthermore no cross reactivity was observed for the nanoPCR assay with respect to related viruses, including canine distemper virus (CDV) and Aleutian mink disease parvovirus (AMDV). Phylogenetic analysis of four Chinese wild type MEV isolates using the nanoPCR assay indicated that they belonged to a small MEV clade, named "China type", in the MEV/FPLV cluster, and were closely clustered in the same location.
  
  
  CONCLUSIONS
  Our results indicate that the MEV China type clade is currently circulating in domestic minks in China. We anticipate that the nanoPCR assay we have described here will be useful for the detection and epidemiological and pathological characterization of MEV.
authors.affiliation: State Key Laboratory for Molecular Biology of Special Economic Animals, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, 130112, China. tcswjk@126.com.
authors.forename: Jianke
authors.initials: J
authors.lastname: Wang
authors.affiliation: Jilin Teyan Biological Technology Company, Changchun, 130122, China. 89331641@qq.com.
authors.forename: Yuening
authors.initials: Y
authors.lastname: Cheng
authors.affiliation: Jilin Teyan Biological Technology Company, Changchun, 130122, China. 756160768@qq.com.
authors.forename: Miao
authors.initials: M
authors.lastname: Zhang
authors.affiliation: State Key Laboratory for Molecular Biology of Special Economic Animals, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, 130112, China. 1797630148@qq.com.
authors.forename: Hang
authors.initials: H
authors.lastname: Zhao
authors.affiliation: State Key Laboratory for Molecular Biology of Special Economic Animals, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, 130112, China. 740891227@qq.com.
authors.forename: Peng
authors.initials: P
authors.lastname: Lin
authors.affiliation: State Key Laboratory for Molecular Biology of Special Economic Animals, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, 130112, China. tcs_yili@126.com.
authors.forename: Li
authors.initials: L
authors.lastname: Yi
authors.affiliation: State Key Laboratory for Molecular Biology of Special Economic Animals, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, 130112, China. 498183427@qq.com.
authors.forename: Mingwei
authors.initials: M
authors.lastname: Tong
authors.affiliation: State Key Laboratory for Molecular Biology of Special Economic Animals, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, 130112, China. tcscsp@126.com.
authors.forename: Shipeng
authors.initials: S
authors.lastname: Cheng
mesh_terms: D000818:Animals; D002681:China; D015967:Gene Expression Regulation, Viral; D014644:Genetic Variation; D008907:Mink; D053489:Mink Viral Enteritis / Q000453:epidemiology / Q000821:virology*; D053490:Mink enteritis virus / Q000235:genetics* / Q000302:isolation & purification*; D053758:Nanoparticles; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: C100315:NS1 protein, Flavivirus; D017361:Viral Nonstructural Proteins