title: [Rapid detection of Macrobrachium rosenbergii nodavirus isolated in China by a reverse-transcription loop-mediated isothermal amplification assay combined with a lateral flow dipstick method].
abstract: White coloration of the muscle of the giant river prawn (Macrobrachium rosenbergii) is a serious problem in China. The Macrobrachium rosenbergii Nodavirus (MrNV) has been confirmed to be the pathogen that causes this disorder. To develop a rapid, sensitive and specific technology for the detection of Macrobrachium rosenbergii Nodavirus isolated from China (MrNV-China), a reverse-transcription loop- mediated isothermal amplification assay combined with a lateral flow dipstick (RT-LAMP-LFD) assay method is described. A set of four primers and a labeled probe were designed specifically to recognize six distinct regions of the MrNV RNA2 gene. Results showed the sensitivity of the RT-LAMP-LFD assay was ten-times higher than the reverse-transcription loop-mediated isothermal amplification assay (RT-LAMP) with agarose gel electrophoresis. The assay was conducted with one-step amplification at 61°C in a single tube within 45 min. No product was generated from shrimps infected with other viruses, including DNA viruses (infectious hypodermal and hematopoietic necrosis virus (IHHNV); white spot syndrome virus (WSSV)) and RNA viruses (Taura syndrome virus (TSV); infectious myonecrosis virus (IMNV); yellow head virus (YHV)). Results were visualized by the LFD method. Therefore, the described rapid and sensitive assay is potentially useful for MrNV detection.
authors.affiliation: Key Laboratory of Healthy Freshwater Aquaculture, Zhejiang Institute of Freshwater Fisheries, Huzhou, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Lin
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.forename: Gui-Jie
authors.initials: GJ
authors.lastname: Hao
authors.forename: Zheng
authors.initials: Z
authors.lastname: Cao
authors.forename: Peng-Cheng
authors.initials: PC
authors.lastname: Sheng
authors.forename: Ying-Lei
authors.initials: YL
authors.lastname: Wu
authors.forename: Jin-Yu
authors.initials: JY
authors.lastname: Shen
mesh_terms: D000818:Animals; D030381:Nodaviridae / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D033541:Palaemonidae / Q000821:virology*; D048348:Reverse Transcription