title: Rapid detection of peste des petits ruminants virus by a reverse transcription loop-mediated isothermal amplification assay.
abstract: Peste des petits ruminants virus (PPRV) is the causative agent of peste des petits ruminants (PPR), an economically important viral disease of small ruminants. In this report, a one-step, single-tube, reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of PPRV. A set of six LAMP primers were designed based on the matrix gene sequence of PPRV to amplify the target RNA by incubation at 63°C for 60min with Bst DNA polymerase and reverse transcriptase. The amplified products could be observed by the naked eye. The specificity of the RT-LAMP assay was validated by amplifying eight strains of PPRV isolated in different geographical areas. No cross-reactivity with other related viruses, including rinderpest virus, canine distemper virus and measles virus, was detected. The sensitivity of the assay was similar to that of real-time reverse transcription polymerase chain reaction (RT-PCR) and 10-fold higher than that of conventional RT-PCR. Twenty clinical samples were evaluated by the RT-LAMP assay, and the results were consistent with those of real-time RT-PCR. As a simple, rapid and accurate detection method, this RT-LAMP assay has important potential applications in the clinical diagnosis of PPR and the surveillance of PPRV.
authors.affiliation: China Animal Health and Epidemiology Center, Qingdao, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Li
authors.forename: Jingyue
authors.initials: J
authors.lastname: Bao
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Wu
authors.forename: Zhiliang
authors.initials: Z
authors.lastname: Wang
authors.forename: Junwei
authors.initials: J
authors.lastname: Wang
authors.forename: Mingxia
authors.initials: M
authors.lastname: Gong
authors.forename: Chunju
authors.initials: C
authors.lastname: Liu
authors.forename: Jinming
authors.initials: J
authors.lastname: Li
mesh_terms: D000818:Animals; D001483:Base Sequence; D003429:Cross Reactions; D004587:Electrophoresis, Agar Gel; D005456:Fluorescent Dyes; D015511:Goat Diseases / Q000175:diagnosis / Q000821:virology; D006041:Goats / Q000821:virology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D029021:Peste-des-Petits-Ruminants / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D018107:Peste-des-petits-ruminants virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D012756:Sheep / Q000821:virology; D012757:Sheep Diseases / Q000175:diagnosis / Q000821:virology; D013194:Staining and Labeling / Q000379:methods
chemical_list: D005456:Fluorescent Dyes; D012367:RNA, Viral