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Method for detecting oyster ploidy by using lymphocyte in oyster coelomic fluid

A technology of lymphocytes and body cavity fluid, which is applied in the field of shellfish polyploid detection, can solve problems such as unfavorable sample application, and achieve a highly practical effect

Active Publication Date: 2015-08-05
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This sampling operation kills the sample itself, which is not conducive to the later application of the sample

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] a. Lymph fluid extraction: In November 2014, 100 Hong Kong oyster holotriploid samples with a shell height of 50-80 mm were retrieved from Hengqin Island, Zhuhai, and sent to the Key Point of Tropical Marine Biological Resources and Ecology of the South China Sea Institute of Oceanology, Chinese Academy of Sciences In the laboratory, first number them, and then place them in seawater with a temperature of 25-27°C and a salinity of 18ppt. After 5 minutes, all oysters open their double shells slightly. When the oysters naturally open their double shells, use a syringe to gently insert the Hong Kong oyster In the body cavity, when the oyster feels the needle piercing, it will close the two shells tightly, but it will not affect the extraction of the body cavity fluid. Each oyster extracts 1.0ml of body cavity fluid, numbered and placed in a centrifuge tube;

[0021] b. Centrifugal resuspension: Centrifuge the body cavity fluid in the centrifuge tube at 1000r / min, remove the...

Embodiment 2

[0025] a. Lymph fluid extraction: In December 2014, 50 Hong Kong oyster betatriploid mixed samples with a shell height of 50-80 mm were retrieved from Zhuhai to the Zhanjiang Marine Economic Animal Experimental Station of the South China Sea Institute of Oceanology, Chinese Academy of Sciences. After 8 minutes, all the oysters opened their double shells slightly. When the oysters opened their double shells naturally, use a syringe to gently insert them into the body cavity of Hong Kong oysters. When the oysters When the needle is felt, the two shells will be closed tightly, but the extraction of body cavity fluid will not be affected. Each oyster will extract 0.8mL of body cavity fluid, numbered and placed in a centrifuge tube;

[0026] b. Centrifugal resuspension: Centrifuge the body cavity fluid in the centrifuge tube at 800r / min, remove the supernatant after centrifugation, add 0.5ml of seawater, blow gently with a syringe for secondary suspension, and then filter with a fil...

Embodiment 3

[0030] a. Lymph fluid extraction: In December 2014, 50 Hong Kong oyster betatriploid mixed samples with a shell height of 30-60 mm were retrieved from the North Sea to the Zhanjiang Marine Economic Animal Experimental Station of the South China Sea Institute of Oceanology, Chinese Academy of Sciences. After 10 minutes, all the oysters opened their double shells slightly. When the oysters naturally opened their double shells, use a syringe to gently insert them into the body cavity of Hong Kong oysters. When the oysters When the needle is felt, the two shells will be closed tightly, but the extraction of body cavity fluid will not be affected. 0.5mL of body cavity fluid will be extracted from each oyster, numbered and placed in a centrifuge tube;

[0031] b. Centrifugal resuspension: centrifuge the body cavity fluid in the centrifuge tube at 1200r / min, remove the supernatant after centrifugation, add 0.5ml of seawater, blow gently with a syringe for secondary suspension, and the...

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Abstract

The invention discloses a method for detecting oyster ploidy by using lymphocyte in oyster coelomic fluid. According to the method, the lymphocyte in the oyster sample coelomic fluid is used as a detection material; the content of DNA (Deoxyribonucleic Acid) is detected; then the ploidy composition of the oyster sample is judged. The method is different from the existing oyster ploidy determination operation; in the earlier detection, tissues such as gill, mantle and adductor muscle are required to be adopted as samples, only single-cell suspension is required to be prepared, and the samples are also required to be killed, so the subsequent work of the samples is difficult to develop. By combining the characteristics that the oyster has an open vascular cycle and the coelomic fluid contains a large amount of lymphocytes, only the single-cell lymphocyte in the coelomic fluid is adopted as the detection sample, so the single-cell suspension preparation step is omitted, and the detection accuracy is 100 percent, 100 percent survival of oyster samples can be guaranteed, and the oyster samples are not damaged, so a theoretical and practical application foundation is laid for in vivo detection of the oyster ploidy. The method has the advantages of quickness, easiness, convenience, high practicability and the like.

Description

technical field [0001] The invention belongs to the technical field of shellfish polyploid detection in marine agriculture, and in particular relates to a method for detecting oyster ploidy by using lymphocytes in oyster body cavity fluid. Background technique [0002] The circulatory system of animals has two types: open-tube circulation and closed-tube circulation. The blood in the open-tube circulation flows out through the heart and large blood vessels, and then enters the inter-organ or interstitial space, where the blood directly contacts with its cells. The blood of the closed-tube circulation always flows in the blood vessels, and the blood does not directly contact with its cells (Xu Junyan, Dong Xiuli, 1998). For molluscs, the body cavity evolves into a form of coexistence of true and false coelom, the true coelom degenerates into traces, and remains in the pericardial cavity and the official cavity of reproductive organs and excretory organs, while the false coelo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 李军张扬张跃环喻子牛
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI