title: Development of a reverse-transcription loop-mediated isothermal amplification method for detection of rabbit hemorrhagic disease virus.
abstract: Rabbit hemorrhagic disease virus (RHDV) causes haemagglutination and severe liver damage, with a high mortality rate. To develop a rapid and sensitive method for the surveillance of RHDV, a one-step reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay was established using a set of four primers specific for the VP60 gene segment of RHDV. The established assay was performed at 64°C for 40 min under isothermal conditions, and the results were visualized directly by electrophoresis or as fluorescent signals under ultraviolet light. The detection limit of the RT-LAMP assay was 10 copies of viral RNA per reaction, which was comparable to quantitative real-time RT-PCR, and 100-fold more sensitive than standard RT-PCR. Furthermore, seven viral RNAs of field isolates in China could be detected successfully using this assay. Overall, the newly established RT-LAMP assay indicates the potential usefulness of the technique as a simple, rapid and sensitive procedure, and can visually detect RHDV infection without the need for any specialized equipment.
authors.affiliation: Experimental Animal Research Center, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, China.
authors.forename: Dongwei
authors.initials: D
authors.lastname: Yuan
authors.forename: Dongchun
authors.initials: D
authors.lastname: Guo
authors.forename: Jiasen
authors.initials: J
authors.lastname: Liu
authors.forename: Changde
authors.initials: C
authors.lastname: Si
authors.forename: Qian
authors.initials: Q
authors.lastname: Jiang
authors.forename: Huan
authors.initials: H
authors.lastname: Lin
authors.forename: Tiankuo
authors.initials: T
authors.lastname: Yang
authors.forename: Liandong
authors.initials: L
authors.lastname: Qu
mesh_terms: D000818:Animals; D017250:Caliciviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D002681:China; D017931:DNA Primers / Q000235:genetics; D017929:Hemorrhagic Disease Virus, Rabbit / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011817:Rabbits; D012376:Rodent Diseases / Q000175:diagnosis* / Q000821:virology*; D012680:Sensitivity and Specificity; D013696:Temperature; D013997:Time Factors; D014730:Veterinary Medicine / Q000379:methods*; D014773:Virology / Q000379:methods*
chemical_list: D017931:DNA Primers