LAMP detection kit for macrobrachium rosenbergii nudivirus and detection method thereof

A detection kit, Macrobrachium rosenbergii technology, applied in the direction of microbe-based methods, microbiological determination/inspection, biochemical equipment and methods, etc., can solve problems such as hindering disease prevention, immeasurable economic losses, and lack of detection methods , to achieve the effect of overcoming long detection time, reducing background influence, and high yield

Active Publication Date: 2012-11-28
ZHEJIANG INST OF FRESH WATER FISHERIES
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Problems solved by technology

The virus was discovered in 2010 from Macrobrachium rosenbergii larvae suffering from Macrobrachium rosenbergii larval metamorphosis syndrome, which caused a large number of deaths of Macrobrachium rosenbergii in the larval stage, especially in the late stage of larvae and the process of metamorphosis into larvae. The mortality rate is generally 70-80%, and it can reach more than 90% in severe cases. The economic losses caused in the past 2 years are immeasurable
Since it was discovered, due to the lack of detection methods, the prevention of diseases caused by the virus has been seriously hindered, so the development of the virus detection kit and the research of detection technology are particularly important

Method used

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  • LAMP detection kit for macrobrachium rosenbergii nudivirus and detection method thereof
  • LAMP detection kit for macrobrachium rosenbergii nudivirus and detection method thereof

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Embodiment Construction

[0045] The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.

[0046] (1) DNA extraction:

[0047] Take 0.1-0.2 g of the cephalothorax of Macrobrachium rosenbergii larvae after removing the eyes or the gill tissue of adult shrimp, grind it with a grinding rod on ice, add 400 μL TE buffer solution (containing 5-15 mM Tris, 0.5-1.5 mM EDTA, pH 8.0) and 1 μL of RNase A with a concentration of 10 mg / mL, continue to grind sufficiently, then add 20 μL of proteinase K solution with a concentration of 20 mg / mL and a pH value of 8.0, vortex for 1 minute, bathe in 56 ° C for 30 min, add the same volume of pH value 8.0 Tris saturated phenol, shake vigorously, centrifuge at 11000g for 3 minutes, take the supernatant, and repeat the phenol extraction; take the supernatant, add 0.1 times the volume of sodium acetate with a concentration of 3mol / L, mix well, and add an equal volume After mixing, let stand at low temper...

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Abstract

Provided are an LAMP (loop-mediated isothermal amplification) detection kit for macrobrachium rosenbergii nudivirus and a detection method thereof. The invention belongs to the technical field of DNA fragment detection. The LAMP detection kit comprises a viral DNA extraction reagent and an LAMP reaction reagent. The LAMP reaction reagent contains an LAMP amplification-used nucleic acid primer group for detecting the macrobrachium rosenbergii nudivirus. The primer group contains a primer of MRNuV-FIP, a primer of MRNuV-BIP, a primer of MRNuV-F3, a primer of MRNuV-B3, a primer of MRNuV-LF and a primer of MRNuV-LB. The kit of the invention is high in specificity and sensitivity. The LAMP detection method for macrobrachium rosenbergii nudivirus makes use of the kit, and is convenient, sensitive, accurate and rapid.

Description

technical field [0001] The invention relates to the technical field of detection of target DNA fragments, in particular to a kit and a detection method for rapid detection of Macrobrachium rosenbergii Noti virus by using a loop-mediated isothermal amplification (Loop-mediated isothermal amplification, LAMP) technology. Background technique [0002] Macrobrachium rosenbergii Nudivirus (MRNuV) is one of the main pathogens causing larval metamorphosis syndrome of Macrobrachium rosenbergii, which is extremely harmful to the nursery industry of Macrobrachium rosenbergii. MRNuV is a double-stranded DNA virus belonging to the genus Notivirus, and the classification status of this genus has not been clarified internationally. The virus was discovered in 2010 from Macrobrachium rosenbergii larvae suffering from Macrobrachium rosenbergii larval metamorphosis syndrome, which caused a large number of deaths of Macrobrachium rosenbergii in the larval stage, especially in the late stage o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 潘晓艺沈锦玉蔺凌云姚嘉赟徐洋袁雪梅郝贵杰尹文林
Owner ZHEJIANG INST OF FRESH WATER FISHERIES
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