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Prawn white spot complex virogene diagnostic kit and detecting method thereof

A technology for leukoplakia syndrome and genetic diagnosis, which is applied in biochemical equipment and methods, and microbial measurement/inspection, etc., can solve problems such as difficulty in popularization, troublesome material preparation, and low sensitivity, so as to improve scientific management efficiency and avoid virus transmission Popular, Fast Effects Guaranteed

Inactive Publication Date: 2003-10-15
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Some of these detection methods have high technical requirements, long detection time, troublesome preparation of materials required, high cost, and low sensitivity and low specificity of some methods. Therefore, it is very difficult for the majority of shrimp farmers to master key technologies, and it is difficult to popularize , limiting the application and development of these technologies

Method used

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  • Prawn white spot complex virogene diagnostic kit and detecting method thereof
  • Prawn white spot complex virogene diagnostic kit and detecting method thereof

Examples

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Effect test

Embodiment 1

[0024] Embodiment 1: the genetic diagnosis kit of prawn white spot syndrome virus

[0025] The kit consists of the following components (10 samples): 1. Sample diluent (solution A), 1 tube, 5ml / tube, containing phosphate buffer (1×PBS), pH7.4. 2. Template extraction solution ( Solution B), 1 tube, 10 μl / tube, filled with proteinase K. 3. PCR reaction solution (solution C), 1 tube, 250μl / tube, containing PCR first expansion reaction solution (25μl system), including ddH 2 O, 10×Buffer (including mg 2+ ), dNTP, outer primer F1, outer primer R1 and TaqE. 4. PCR reaction solution (solution D), 1 tube, 250μl / tube, containing PCR secondary amplification reaction solution (25μl system), including ddH 2 O, 10×Buffer (including mg 2+ ), dNTP, inner primer F2, inner primer R2 and TaqE. 5. Positive control (solution E), 1 tube, 20 μl / tube, containing WSSV positive DNA. 6. Cuboid box, 8.5×5.8×6.2cm 3 . 7. A piece of foam board whose size is the same as the bottom surface of the cu...

Embodiment 2

[0046] Embodiment 2: the detection method of prawn white spot syndrome virus

[0047] Using the kit of Example 1, proceed as follows: 1. Take 0.05 g of the sample, add 500 ul of sample diluent (liquid A) to dilute 10 times, and homogenize in an ice bath in a homogenizer. 2. Centrifuge at 6000r / min for 5min. 3. Take 50 μl supernatant and add 1 μl template extract solution (solution B), boil for 15 minutes, and immediately put it on ice for 5 minutes. 4. Centrifuge at 8000r / min for 10min, and use the supernatant as a PCR template. 5. Take 2 μl of the template and solution E respectively, add them to the PCR reaction solution (solution C), mix well, centrifuge for a few seconds, and place on the PCR instrument. 6. Amplify according to the following conditions:

[0048] Pre-denaturation at 95°C for 2min→34 cycles at 94°C for 45sec→10min at 72°C→storage at 4°C

[0049] 55℃ 45sec

[0050] 72°C for 1min7. Take 2μl of the first amplification reac...

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Abstract

The present invention is prawn white spot syndrome virus gene diagnosing kit and its usage. The kit and its usage is designed based on two pairs of primers for the conservation sequence of prawn white spot syndrome virus gene. By means of PCR technology, the present invention detects qualitatively the specific DNA nucleic acid segment of prawn white spot syndrome virus fast, simply, specifically and sensitively. The present invention may be used in the tracing detection of different stages of prawn cultivation and environment monitoring.

Description

technical field [0001] The invention relates to a diagnostic kit and a detection method for diseases of aquatic economic animals, and mainly relates to a kit and a detection method for genetic diagnosis of white spot syndrome virus (WSSV, White Spot Syndrome Virus) of prawns. Background technique [0002] Since the 1990s, shrimp diseases have exploded in many countries and regions in Asia, and the production of prawn farming has dropped sharply, causing major economic losses. Shrimp diseases, especially shrimp virus diseases, have seriously hindered the sustainable development of the shrimp farming industry. Among the many shrimp viruses, the white spot syndrome virus (WSSV) is the most serious, and it is the number one killer of the shrimp farming industry. The research on WSSV is also a hot spot in the research of aquaculture diseases in recent years. At present, there is no specific treatment method for shrimp virus disease. The most effective preventive measure is to p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/70
Inventor 何建国邓敏黄志坚吕玲翁少萍
Owner SUN YAT SEN UNIV
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