Prawn taura syndrome virogene diagnostic kit and detecting method

A syndrome virus and gene diagnosis technology, which is applied in the field of prawn taura syndrome virus gene diagnosis kit and detection, can solve problems such as difficult promotion, troublesome material preparation, and low sensitivity, so as to improve scientific management efficiency and avoid virus transmission Popular, Fast Effects Guaranteed

Inactive Publication Date: 2005-02-16
SUN YAT SEN UNIV +1
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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

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  • Prawn taura syndrome virogene diagnostic kit and detecting method

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Embodiment 1

[0040] Embodiment 1: the genetic diagnosis kit of prawn taura syndrome virus

[0041] The kit consists of the following parts (10 samples):

[0042] 1. RNA extraction solution (solution A), 2 tubes, 5ml / tube, filled with Trizol solution.

[0043] 2. Solution B, 1 tube, filled with chloroform (you can also bring your own), 2ml in total (200μl / part x 10 parts).

[0044] 3. Liquid C, 1 tube, filled with isopropanol (you can also bring your own), 5ml in total (500μl / part x 10 parts).

[0045] 4. Solution D, 1 tube, filled with 70% ethanol (you can also bring your own), 10ml in total (1ml / part x 10parts).

[0046] 5. Liquid E, 1 tube, 0.5ml / tube, filled with DEPC water.

[0047] 6. RT reaction solution (solution F), 1 tube, 0.1ml / tube, containing RT reaction solution (10μl system), including 5×Buffer, dNTP, DEPC-H 2 O, random primers, RNase inhibitor RNAsin (RI), reverse transcriptase M-MLV (RT).

[0048] 7. RT-PCR reaction solution (solution G), 1 tube, 0.5ml / tube, containing...

Embodiment 2

[0069] Embodiment 2: the detection method of prawn taura syndrome virus

[0070] Use the test kit of embodiment 1, carry out according to the following steps:

[0071] 1. Using aseptic operation method, take 0.1g of fresh prawn liver tissue and add 1ml of liquid A to dilute, homogenize in ice bath in a homogenizer, and let it stand at room temperature for 3-5min.

[0072] 2. Add 200μl B solution to the above homogenate, mix by inverting up and down, and let it stand for 5 minutes.

[0073] Centrifuge at 12000r / min for 15min at 3.4°C.

[0074] 4. Take 400 μl supernatant, add it to 500 μl solution C, shake gently, and let it stand for 10 minutes.

[0075] Centrifuge at 12000r / min for 15min at 5.4°C.

[0076] 6. Discard the supernatant, wash twice with 1ml of pre-cooled solution D, and centrifuge at 7500r / min for 5min at 4°C.

[0077] 7. Air dry or blow dry on a clean bench, add 50μl E liquid water to dissolve (if it cannot be completely dissolved, it can be placed at 55-60°C...

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Abstract

The invention provides a kind of gene consultation kit and testing process to prawn TSV( Taura Syndrome virus). The kit and the testing process are designed based on list in conservation area of TSV. The invention adopts RT-PCR technology to test special RNA nucleic acids of TSV. This method is quick and its specificity is good and sensitivity is high. It can be used to test virus in different period when cultivate prawn. It also can be used to test environment to avoid dissmination of virus. Thus, it can improve efficiency of scientific management and has high utility 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 kit and a detection method for gene diagnosis of prawn Taura Syndrome Virus (TSV, Taura Syndrome Virus). 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, Shrimp Taura Syndrome Virus (TSV) is more serious, and it is the number one killer of Penaeus vannamei aquaculture. The research on TSV 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 promote healthy shrimp farming,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12Q1/68
Inventor 何建国黄志坚翁少萍陆辉
Owner SUN YAT SEN UNIV
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