title: Establishment of reverse transcription loop-mediated isothermal amplification for rapid detection and differentiation of canine distemper virus infected and vaccinated animals.
abstract: Although widespread vaccination against canine distemper virus (CDV) has been conducted for many decades, several canine distemper outbreaks in vaccinated animals have been reported frequently. In order to detect and differentiate the wild-type and vaccine strains of the CDV from the vaccinated animals, a novel reverse transcription loop-mediated isothermal amplification (RT-LAMP) method was developed. A set of four primers-two internal and two external-were designed to target the H gene for the specific detection of wild-type CDV variants. The CDV-H RT-LAMP assay rapidly amplified the target gene, within 60 min, using a water bath held at a constant temperature of 65°C. The assay was 100-fold more sensitive than conventional RT-PCR, with a detection limit of 10(-1)TCID50ml(-1). The system showed a preference for wild-type CDV, and exhibited less sensitivity to canine parvovirus, canine adenovirus type 1 and type 2, canine coronavirus, and canine parainfluenza virus. The assay was validated using 102 clinical samples obtained from vaccinated dog farms, and the results were comparable to a multiplex nested RT-PCR assay. The specific CDV-H RT-LAMP assay provides a simple, rapid, and sensitive tool for the detection of canines infected with wild-type CDV from canines vaccinated with attenuated vaccine.
authors.affiliation: College of Wildlife Resources, Northeast Forestry University, Harbin 150040, China; Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China.
authors.forename: Da-Fei
authors.initials: DF
authors.lastname: Liu
authors.affiliation: Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China.
authors.forename: Chun-Guo
authors.initials: CG
authors.lastname: Liu
authors.affiliation: Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China. Electronic address: 289533973@qq.com.
authors.forename: Jin
authors.initials: J
authors.lastname: Tian
authors.affiliation: Beijing Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100091, China.
authors.forename: Yi-Tong
authors.initials: YT
authors.lastname: Jiang
authors.affiliation: Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China.
authors.forename: Xiao-Zhan
authors.initials: XZ
authors.lastname: Zhang
authors.affiliation: College of Wildlife Resources, Northeast Forestry University, Harbin 150040, China.
authors.forename: Hong-Liang
authors.initials: HL
authors.lastname: Chai
authors.affiliation: Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China.
authors.forename: Tian-Kuo
authors.initials: TK
authors.lastname: Yang
authors.affiliation: College of Wildlife Resources, Northeast Forestry University, Harbin 150040, China.
authors.forename: Xiu-Chen
authors.initials: XC
authors.lastname: Yin
authors.affiliation: Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China.
authors.forename: Hong-Ying
authors.initials: HY
authors.lastname: Zhang
authors.affiliation: Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Liu
authors.affiliation: College of Wildlife Resources, Northeast Forestry University, Harbin 150040, China. Electronic address: 112441457@qq.com.
authors.forename: Yu-Ping
authors.initials: YP
authors.lastname: Hua
authors.affiliation: Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China. Electronic address: qld@hvri.ac.cn.
authors.forename: Lian-Dong
authors.initials: LD
authors.lastname: Qu
mesh_terms: D000818:Animals; D004216:Distemper / Q000175:diagnosis / Q000517:prevention & control / Q000821:virology*; D004217:Distemper Virus, Canine / Q000235:genetics* / Q000276:immunology; D004285:Dogs / Q000821:virology; D060885:Multiplex Polymerase Chain Reaction / Q000662:veterinary; D021141:Nucleic Acid Amplification Techniques / Q000379:methods / Q000662:veterinary*; D017993:Parvovirus, Canine / Q000235:genetics; D012680:Sensitivity and Specificity; D014765:Viral Vaccines / Q000235:genetics* / Q000627:therapeutic use
chemical_list: D014765:Viral Vaccines
keywords: Attenuated vaccine; Canine distemper virus (CDV); Differential diagnosis; Loop-mediated isothermal amplification (LAMP)