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Rapid detection kit and detection method of Chinese softshell turtle artertivirus

A technology of Arteritis virus and detection kit, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, and microorganism measurement/inspection, and can solve problems such as hindering disease prevention, death of Chinese soft-shelled turtle, and lack of detection methods. Achieve the effects of overcoming long detection time, high yield, and reducing background influence

Inactive Publication Date: 2016-07-20
ZHEJIANG INST OF FRESH WATER FISHERIES
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Problems solved by technology

The virus was isolated from soft-shelled soft-shelled turtles suffering from mumps and pneumonia in 2012. It caused a large number of soft-shelled turtles to die during the breeding period. The mortality rate was generally 50% to 70%, and the severe cases reached more than 80%. The economic loss is 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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  • Rapid detection kit and detection method of Chinese softshell turtle artertivirus
  • Rapid detection kit and detection method of Chinese softshell turtle artertivirus
  • Rapid detection kit and detection method of Chinese softshell turtle artertivirus

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Embodiment 1

[0037] (1) RNA extraction:

[0038] Take 30-80 mg of soft-shelled turtle spleen or heart tissue in a 2 mL centrifuge tube, grind it on ice with a grinding rod, add 600 μL of lysate A, continue grinding, add 600 μL of Tris saturated phenol with a pH value of 8.0, shake vigorously, and centrifuge at 11,000 g for 10 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 3mol1 / L, mix well, add two times the volume of ice-cold absolute ethanol, mix well and let it cool down Set aside for 10 minutes, centrifuge at 15,000 g for 5 minutes, discard the supernatant, wash the precipitate twice with 75% ethanol, dry at room temperature for 5-10 minutes, resuspend in 30 μL DEPC water, and store at -80°C for future use.

[0039] (2) Rapid amplification of Chinese soft-shelled turtle arteritis virus:

[0040]According to the number of samples to be detected, the required rapid reaction tube nu...

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Abstract

The invention discloses a rapid detection kit and detection method of Chinese softshell turtle artertivirus.The rapid detection kit of Chinese softshell turtle artertivirus comprises virus RNA extraction reagent and reaction reagent, the reaction reagent comprises a nucleic acid primer group for detecting amplification of Chinese softshell turtle artertivirus, and the primer group comprises a primer TSAV-F3, a primer TSAV-B3, a primer TSAV-FIP and a primer TSAV-BIP.By means of the rapid detection kit, an optimized ring mediated isothermal amplification reaction system is established, qualitative detection of Chinese softshell turtle artertivirus is made to be easier, more convenient and more rapid, specificity is high, sensitivity is high, the kit is the first kit for detecting Chinese softshell turtle artertivirus, the blank that no method for detecting Chinese softshell turtle artertivirus exists is made up for, and quite high scientific research and economic value is achieved.

Description

technical field [0001] The invention belongs to the field of rapid detection of target RNA fragments, and in particular relates to a rapid detection kit for soft-shelled turtle arteritis virus and a detection method thereof. Background technique [0002] Chinese soft-shelled turtle arteritis virus (Trionyxsinensis Arterivirus, TSAV) is one of the main pathogens causing viral pneumonia and arteritis of Chinese soft-shelled turtle, which is extremely harmful to the breeding industry of Chinese soft-shelled turtle. TSAV is a single positive-strand RNA virus belonging to the Arteriviridae family of the order Neviridae, and it is the second RNA virus found to be able to infect Chinese soft-shelled turtles. The virus was isolated from soft-shelled soft-shelled turtles suffering from mumps and pneumonia in 2012. It caused a large number of soft-shelled turtles to die during the breeding period. The mortality rate was generally 50% to 70%, and the severe cases reached more than 80%....

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/701C12Q1/6844C12Q2531/119C12Q2537/1376
Inventor 潘晓艺蔺凌云袁雪梅沈锦玉徐洋姚嘉赟尹文林
Owner ZHEJIANG INST OF FRESH WATER FISHERIES
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