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

A technique for rapid diagnosis of abalone muscular dystrophy, applied in the field of diagnostic kits for aquatic economic animal diseases, to achieve good specificity, improve scientific management efficiency, and speed up the detection process

Inactive Publication Date: 2012-12-05
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
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Effect test

Embodiment 1

[0061] Embodiment 1: the genetic diagnosis kit of abalone muscular dystrophy virus

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

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

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

[0065] ② Solution B: 1 bottle, 20mL / bottle, ingredients: 12mL of 5mol...

Embodiment 2

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

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

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

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

[0093] d) Add 200 μL of solution B to the solution obtained in step c), shake vigorously for 20 seconds; then centrifuge at 12,000 r / min for 3 minutes at 4°C, take 400 μL of supernatant, add an equal volume of solution C, and shake gently to mix; Centrifuge at 12000r / min for 3min at 4°C, discard the supernatant, wash the resulting precipitate once with 600μL pre-co...

Embodiment 3

[0103] The difference with embodiment 2 is:

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

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

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

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

[0108] d) Add 100 μL of solution B to the solution in step c), shake vigorously for 10 seconds; then centrifuge at 13,000 r / min for 1 min at 4°C, take 200 μL of supernatant, add an equal volume of solution C, and shake gently to mix; Centrifuge at 13000r / min for 1min at ℃, discard the supernatant, wash the obtained prec...

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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 aquatic economic animal diseases, in particular to a rapid diagnostic kit and a detection method for abalone muscular dystrophy virus. Background technique [0002] Abalone is a traditional and precious food material in China. Its meat is delicious and nutritious, and it is a marine economic shellfish widely cultivated all over the world. In the winter of 2005, large-scale disease outbreaks occurred in abalone farms in Fujian, Guangdong and other places. The main symptoms were muscle and mantle atrophy and black body color. Abalone can be infected at any stage, 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 muscular dystrophy virus (Abalone Shriveling syndrome associated Virus, AbSV). So far, there is no effective treatment for viral diseases. Preventing the infection and prevalen...

Claims

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Application Information

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Patent Type & Authority Patents(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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