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Lamp detection kit and detection method of macrobrachium rosenbergii dicistronic virus

A detection kit, Macrobrachium rosenbergii technology, applied in the detection field of target DNA fragments, can solve the problems of death, lack of detection methods, hindering disease prevention, etc., achieve high amplification efficiency, overcome long detection time, and reduce background influence Effect

Active Publication Date: 2011-12-14
ZHEJIANG INST OF FRESH WATER FISHERIES
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AI Technical Summary

Problems solved by technology

The virus caused the disease of Macrobrachium rosenbergii larvae in my country in 2009 and 2010, causing a large number of deaths in the larval stage of Macrobrachium rosenbergii, especially when the larvae were 6-9 days old, the death rate was particularly serious, and the mortality rate was generally 80-90%. Seriously reaching 100%, causing incalculable economic losses in the past 2 years
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 and detection method of macrobrachium rosenbergii dicistronic virus

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Embodiment 1

[0056] 1) Extract RNA with RNA extraction reagent (RNA extraction reagent is a conventional reagent in the prior art, which contains Trizol reagent, chloroform, isopropanol, RNase-free 70% ethanol and DEPC water):

[0057] Take 0.1, 0.15 or 0.2 g of the cephalothorax of Macrobrachium rosenbergii larvae or the gill tissue of adult shrimp, grind it on ice with a grinding rod, add 300 μL Trizol, continue to grind thoroughly, add Trizol to 1 mL, vortex for 1 minute, and room temperature for 5 minutes , add 200 μL of chloroform, vortex for 30 seconds, stand at room temperature for 3 minutes, centrifuge at 12,000 g for 10 minutes, take the upper aqueous phase into a new EP tube, add 500 μL of isopropanol, leave at room temperature for 10 minutes, and centrifuge at 12,000 g for 10 minutes; remove The supernatant and the precipitate were washed twice with 70% ethanol, dried at room temperature for 5, 8 or 10 minutes, then resuspended with DEPC-treated deionized water, and divided to ob...

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Abstract

The invention discloses an LAMP detection kit and an LAMP detection method for macrobrachium rosenbergii bicistronic virus, which belongs to the technical field of target DNA segments. The detection kit comprises an RNA extraction reagent and a reverse transcription (RT)-LAMP reaction reagent. The detection method comprises: 1) extracting RNA of macrobrachium rosenbergii; 2) performing the RT-LAMP amplification of the macrobrachium rosenbergii bicistronic virus; and 3) performing color developing detection. The detection kit has high specificity and sensitivity, the detection method is convenient, sensitive, accurate and quick, and thus, a foundation is laid for the prevention of syndrome in larvae of macrobrachium rosenbergii.

Description

technical field [0001] The invention belongs to the technical field of detection of target DNA fragments, and in particular relates to a kit and a detection method for rapid detection of Macrobrachium rosenbergii bicistronic virus by using a loop-mediated isothermal amplification (LAMP) technology. Background technique [0002] Macrobrachium rosenbergii bicistronic virus ( Macrobrachium rosenbergii dicistrovirus , MRDV) is the cause of Macrobrachium rosenbergii larval syndrome ( Macrobrachium rosenbergii larval Syndrome, MRLS), which is extremely harmful to the Macrobrachium rosenbergii nursery industry. MRDV is a single positive-sense RNA virus classified in the Dicistroviridae family. The virus has not been reported in the literature at home and abroad, and its genome sequence has only disclosed 900 bases. The virus caused the disease of Macrobrachium rosenbergii larvae in my country in 2009 and 2010, causing a large number of deaths in the larval stage of Macrobrachi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68
Inventor 潘晓艺沈锦玉郝贵杰姚嘉赟徐洋尹文林曹铮
Owner ZHEJIANG INST OF FRESH WATER FISHERIES
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