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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 / 病毒学报
Chinese Journal of Virology ; (6): 502-507, 2014.
Article in Chinese | WPRIM | ID: wpr-280336
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.
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Full text: Available Index: WPRIM (Western Pacific) Main subject: Virology / Nucleic Acid Amplification Techniques / Nodaviridae / Palaemonidae / Reverse Transcription / Methods Type of study: Diagnostic study Limits: Animals Language: Chinese Journal: Chinese Journal of Virology Year: 2014 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Virology / Nucleic Acid Amplification Techniques / Nodaviridae / Palaemonidae / Reverse Transcription / Methods Type of study: Diagnostic study Limits: Animals Language: Chinese Journal: Chinese Journal of Virology Year: 2014 Type: Article