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Macrobrachium rosenbergii Nodavirus NASBA-LFD detection method and detection kit thereof

A technology of Nodamura virus and detection kit, which is applied in the field of NASBA-LFD detection method of Macrobrachium rosenbergii Nodamura virus and its detection kit, can solve the problem of insufficient sensitivity, achieve high detection sensitivity, good specificity, and extraction method simple and fast effect

Active Publication Date: 2013-11-27
ZHEJIANG INST OF FRESH WATER FISHERIES
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  • Application Information

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Problems solved by technology

[0007] Another object of the present invention is to provide a detection kit for the above method, which has strong specificity, high sensitivity, simple and fast operation, overcomes the problem of insufficient sensitivity of existing detection methods, and is suitable for the detection of Macrobrachium rosenbergii. The screening and prevention of disease, as well as the parental traceability screening of diseased shrimp seedlings

Method used

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  • Macrobrachium rosenbergii Nodavirus NASBA-LFD detection method and detection kit thereof
  • Macrobrachium rosenbergii Nodavirus NASBA-LFD detection method and detection kit thereof
  • Macrobrachium rosenbergii Nodavirus NASBA-LFD detection method and detection kit thereof

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Embodiment approach

[0078] The shrimp -like shrimp infected with Luo's shrimp shrimp virus virus is collected from a farm in Xihu District, Hangzhou City, Zhejiang Province;Sulfur sulfur glycol, potassium chloride, etc. are purchased from SIGMA; BSA, RNASE inhibitors, T7 RNA polymerase, and NTP are purchased from Fermentas.

[0079] 2. Extraction of Rochena Shrimp Sample virus RNA:

[0080] (1) Take 25mg of Rochenna shrimp and young shrimp or shrimp muscle tissue, add 500 μl of cracking liquid, use a one -time grinding rod to grind, wait for thoroughly, 12000rpm centrifugal 2min, and absorb the liquid to the new centrifugal tube;

[0081] (2) Add the volume of equal volume (with the volume of the upper liquid) to the new centrifugal tube equipped with the liquid liquid, and mix it evenly after 70%of the volume (with the volume of the upper liquid);

[0082] (3) Transfer the mixed liquid into a adsorption column with a glass fibrin membrane, the adsorbent column is placed in the collection tube, 12000...

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Abstract

The invention discloses a Macrobrachium rosenbergii Nodavirus NASBA-LFD (nuclear acid sequence-based amplification-lateral flow dipstick) detection method and a detection kit thereof. The detection method includes: according to a Macrobrachium rosenbergii Nodavirus (MrNV) sequence published by GenBank, screening a conserved region, and designing the primers and probe needed by an MrNV NASBA-LFD reaction system; employing a tissue RNA extraction reagent prepared by the inventor to extract the RNA of a to-be-detected sample, then utilizing the MrNV NASBA reaction system established by the invention to perform detection, and at the end of the reaction, determining the result according to nucleic acid rapid test paper. Compared with common detection technologies of the virus, the method and the kit provided in the invention have the characteristics of simplicity and rapidity, good specificity and high sensitivity, can be used for field service by technical personnel and farmers at the production front line, and are in favor of diagnosis prevention and pathogen blocking of Macrobrachium rosenbergii white tail disease.

Description

Technical field [0001] The present invention involves the field of aquatic animal virus testing, and specially involves a Rochenuma Shrimpya village virus NASBA-LFD detection method and its detection kit. Background technique [0002] White Tail Disease (WTD), also known as white tail disease, is a Macrobrachium virus (Macrobrachium rosenbergii Nodavirus, mRNV) is a kind of infectious diseases characterized by white spots, white turbidity or whiteness of the whole body with disease shrimp muscles, with a mortality of up to 100%, which has a maximum of 100%of the diseased shrimp muscles.Great harm.At present, WTD is a worldwide Rochenia shrimp disease, and the disease has been listed as OIE application of epidemic and aquatic animal epidemic disease in my country. [0003] Studies have shown that Roche Nona Shrimp Nomota virus particles are uniform and symmetrical 20-facial structures, with smooth surfaces, no capsules, and 26-27nm in diameter.They are AY222839, AY222840).At prese...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 林锋沈锦玉刘莉钱冬曹铮王军毅郝贵杰盛鹏程袁雪梅吴颖蕾
Owner ZHEJIANG INST OF FRESH WATER FISHERIES
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