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Fast quantitative detection method of apostichopus japonicas coelomic fluid virus

A quantitative detection method and detection method technology, applied in the field of aquatic product quality detection, can solve problems such as complicated operation, low feasibility, and long time, and achieve the effect of simple operation

Inactive Publication Date: 2014-08-06
LIAONING OCEAN & FISHERIES SCI RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At this stage, there is still a lack of relevant reports on the types of viruses in sea cucumbers at home and abroad, which also brings great difficulties to the quantitative detection of viruses based on types
In addition, the traditional electron microscope negative staining technology and ultra-thin section technology to detect viruses in the imitation japonicus have disadvantages such as complicated operation, long time, high cost, and low feasibility, and cannot meet the needs of imitation japonicus breeding and production

Method used

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  • Fast quantitative detection method of apostichopus japonicas coelomic fluid virus

Examples

Experimental program
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Effect test

Embodiment 1

[0022] The body cavity fluid virus detection was carried out on the seed ginseng bred by the imitation japonicus seedling breeding center of the Institute of Marine and Fishery Sciences of Liaoning Province. The seed ginseng was randomly selected, a total of 50 imitation japonicus, and the virus was detected from the body cavity fluid of 4 imitation japonicus japonicus . Taking the detection of imitation japonicus ginseng sample a as an example, the specific method is as follows:

[0023] 1. Body cavity fluid extraction

[0024] The body of the imitation japonicus a sample to be tested is about 20 cm in length. Place the imitation japonicus to be tested flat on a clean filter paper, and use the filter paper to blot the seawater on the surface of the imitation japonicus abdomen. The needle of the bacterial syringe was slowly inserted into the abdominal cavity 2 cm above the excretory hole of sea cucumber, and after entering the abdominal cavity 1 cm, slowly extracted 2 mL of b...

Embodiment 2

[0053] The body cavity fluid virus detection was carried out on the imitation sea cucumber seed ginseng of Dalian Wudao Haizhenpin Technology Development Co., Ltd., a total of 10 imitation sea cucumbers were randomly selected, and the virus was detected in the body cavity fluid of 1 imitation sea cucumber. Taking the detection of imitation japonicus ginseng sample b as an example, the specific method is as follows:

[0054] 1. Body cavity fluid extraction

[0055] The body length of japonicus japonicus sample b to be tested is about 15 cm. Put the japonicus japonicus to be tested flat on clean filter paper, and use the filter paper to blot the seawater on the surface of the japonicus japonicus abdomen. The needle of the sterile syringe slowly penetrates the abdominal cavity 1.5cm above the excretory hole of sea cucumber, and after entering the abdominal cavity 0.5cm, slowly draws 1.5mL of body cavity fluid, then slowly pulls out the needle and puts the imitation sea cucumber b...

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Abstract

The invention discloses a fast quantitative detection method of apostichopus japonicas coelomic fluid virus. The method comprises the following steps: 1) extracting apostichopus japonicas coelomic fluid; 2) conducting ultrasonic disruption on coelomic fluid cells, freezing and thawing, and releasing virus particles; 3) filtering the coelomic fluid to prepare a alumina filter membrane containing virus; and 4) staining by avoiding light, flaking, observing and counting. The method provided by the invention can directly and fast detect virus in the apostichopus japonicus without killing the apostichopus japonicas; and the detection process can be completed in 5 hours, has the characteristics of simple operation, fastness and accuracy, and can be used for rapid screening of breeding apostichopus japonicus in apostichopus japonicas larval breeding.

Description

technical field [0001] The invention belongs to the technical field of quality detection of aquatic products, and in particular relates to a rapid quantitative detection method for viruses in body cavity fluid of imitation sea cucumbers. Background technique [0002] my country's imitation japonicus (Apostichopus japonicus) breeding areas are mainly concentrated in the Shandong Peninsula and the coastal areas around the Liaodong Peninsula, with a breeding area of ​​more than one million mu and an annual output value of more than 10 billion yuan. Another new pillar industry. However, the rapid development and non-standard operation of breeding, coupled with the lagging of supporting scientific research, infrastructure, breeding technology and technology, have led to the decline of japonicus japonicus germplasm, frequent occurrence of diseases, and seriously hindered the stability, health and prosperity of the industry. sustainable development. [0003] In recent years, a lar...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12Q1/06G01N21/64C12R1/93
CPCC12Q1/06C12Q2304/10
Inventor 姜北周遵春关晓燕董颖杨爱馥王摆陈仲高杉蒋经伟
Owner LIAONING OCEAN & FISHERIES SCI RES INST
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