Quick diagnostic kit for abalone shriveling syndrome associated virus (AbSV) and detection method

A technology for abalone muscular dystrophy and muscular dystrophy, which is applied in the field of diagnostic kits for aquatic economic animal diseases, and achieves the effects of avoiding virus spread and epidemic, simple and rapid qualitative detection, and speeding up the detection process.

Inactive Publication Date: 2011-09-14
SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no effective treatment for viral diseases. Preventing the infection and prevalence of viruses is the main measure that can be taken. Early and rapid diagnosis of the virus is an effective way to reduce losses. Therefore, it is simple, fast, specific and sensitive. The diagnostic kit and its detection method are of great significance for the prevention of diseases

Method used

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  • Quick diagnostic kit for abalone shriveling syndrome associated virus (AbSV) and detection method
  • Quick diagnostic kit for abalone shriveling syndrome associated virus (AbSV) and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1: Gene diagnosis kit for abalone muscular dystrophy virus

[0062] The kit includes a box and 10 reagent tubes stored in the box. A foam board is placed in the box, the size of which is the same as the bottom of the cuboid box. The foam board has four rows of holes. The first row has four holes with a diameter of 4.3cm. The second row has five holes with a diameter of 3.0cm. There are six holes in four rows, with a diameter of 0.6cm. The above-mentioned small tubes are respectively placed in the holes of the foam board and installed in the rectangular parallelepiped box.

[0063] The reagents contained in the 10 reagent tubes are as follows (50 samples):

[0064] ①A solution: 1 bottle, 40mL / bottle, composition: 10mmol / L Tris-Cl (pH8.0), 1mmol / L EDTA and 0.1% (m / V) SDS mixed solution, as the DNA extraction solution;

[0065] ②B solution: 1 bottle, 20mL / bottle, composition: 12mL 5mol / L potassium acetate, 2.3mL glacial acetic acid and 5.7mL water mixed solution, as the ...

Embodiment 2

[0089] (1) Prepare the DNA template of the abalone sample to be tested;

[0090] a) Take 15mg of fresh abalone variegated muscle tissue sample with sterilized scissors as the sample to be tested, add 600μL A solution to dilute, homogenize in ice bath in a homogenizer to obtain a homogenate;

[0091] b) Add 3 μL of F1 solution to the homogenate of step a), and place it at 55°C for 3 hours;

[0092] c) After the F1 liquid digested homogenate in step b) is cooled to room temperature, 3μL of F2 liquid is added, and it is placed at 37°C for 1 hour for digestion to obtain a digestion solution;

[0093] d) Add 200μL of solution B to the solution obtained in step c), shake vigorously for 20s; then centrifuge at 12000r / min for 3min at 4°C, take 400μL of supernatant, add an equal volume of solution C, shake gently to mix; Centrifuge at 12000r / min for 3min at 4℃, discard the supernatant, and wash the resulting precipitate with 600μL of pre-cooled D solution once, then centrifuge at 12000r / min fo...

Embodiment 3

[0103] The difference from Example 2 is:

[0104] Step (1) Prepare the DNA template of the abalone sample to be tested:

[0105] a) Take 10 mg of fresh abalone mantle sample with sterile scissors as the sample to be tested, add 400μL A solution to dilute, homogenize in an ice bath in a homogenizer to obtain a homogenate;

[0106] b) Add 2μL of F1 solution to the homogenate of step a), and place it at 60°C for 2 hours;

[0107] c) After the F1 liquid digested homogenate in step b) is cooled to room temperature, 2μL of F2 liquid is added, and it is digested at 37°C for 2 hours to obtain a digestion solution;

[0108] d) Add 100μL of solution B to the solution in step c), shake vigorously for 10s; then centrifuge at 13000r / min for 1min at 4°C, take 200μL of supernatant, add an equal volume of solution C, shake gently to mix; Centrifuge at 13000r / min for 1min at ℃, discard the supernatant, and wash the resulting precipitate with 400μL of pre-cooled D solution, then centrifuge at 13000r / min...

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Abstract

The invention discloses a quick diagnostic kit for abalone shriveling syndrome associated virus (AbSV). The quick diagnostic kit comprises the following components: (1) DNA extract; (2) digestive juice; (3) G1 liquid used for polymerase chain reaction (PCR) amplification reaction and comprising Buffer containing Mg<2+>, dNTP, forward primer F1, reverse primer R1 and Taq enzyme, wherein F1 is 5-ATGACAGATTTCACCGTAAGCAATGAAAATC-3, and R1 is 5-CTAGCTGGCTTTGGTATAAGTAGTACCAAAAG-3; and (4) positive control H liquid, which is 100ng/muL of AbSV infected abalone genome DNA. The invention also disclosesa method for detecting the AbSV by using the quick diagnostic kit. The quick diagnostic kit and the detection method provided by the invention can be used for virus tracking detection of each period in the abalone culture process, can also be used for environment monitoring, avoid viral transmission, improve the scientific management efficiency, and have a very high practical value.

Description

Technical field [0001] The invention relates to a diagnostic kit and a detection method for diseases of aquatic economic animals, in particular to a rapid diagnostic kit and a detection method for abalone muscular dystrophy virus. Background technique [0002] Abalone is a traditional precious food in China. Its meat is delicious and nutritious. It is a marine economic shellfish widely cultivated all over the world. In the winter of 2005, large-scale diseases broke out in abalone farms in Fujian, Guangdong and other places. The symptoms were mainly atrophy of muscle and mantle, black body color, and abalone disease at all stages, which caused a devastating blow to the abalone breeding industry in southern my country. . It was later identified that the pathogen of the disease was Abalone Shriveling syndrome associated Virus (AbSV). So far, there is no effective treatment method for viral diseases. Preventing the infection and epidemic of the virus is the main measure that can be ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 姜敬哲王江勇郭志勋庄军刘广锋
Owner SOUTH CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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