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Application of nanogold rod in detection of serum circulating antigen through clonorchis sinensis specific antibody

A nano-gold rod, specific technology, applied in nanotechnology, nanotechnology, nanotechnology for materials and surface science, etc., can solve the problems of inaccurate detection of liver fluke content, low sensitivity, and long time in the early stage

Pending Publication Date: 2020-09-04
DALI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, how to use nano-gold biotechnology to accurately detect liver flukes in the early stage due to low content, and to solve the defects of low sensitivity and long time in the detection methods of liver flukes in the prior art has become a problem that researchers need to face and solve urgently.

Method used

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  • Application of nanogold rod in detection of serum circulating antigen through clonorchis sinensis specific antibody
  • Application of nanogold rod in detection of serum circulating antigen through clonorchis sinensis specific antibody
  • Application of nanogold rod in detection of serum circulating antigen through clonorchis sinensis specific antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] 1. Preparation of liver fluke antiserum and specific IgG antibody

[0043] 1.1 SD rat liver fluke model establishment

[0044] 1.1.1 Collection and isolation of liver fluke metacercariae

[0045] Generally, take 1 to 2 pieces of muscle on the back of the fish (use scissors to pick up the partial fish skin, cut out a piece of soybean-sized fish meat from under the skin, be careful not to have fish skin), put the cut fish meat on the prepared glass On the slice, lightly press the cover glass. For the convenience of observation, make the fish flesh thinner and translucent, and check the metacercariae under the dissecting microscope. Scrape the observed positive fish from the glass slide, and use the prepared digestive juice (digestive juice preparation: absorb 5ml of hydrochloric acid and add it to 1000ml of normal saline, then weigh 7.0g of pepsin at a ratio of 1:3000, and add it to the above mixture , fully mixed, ready to use) It is best to preheat to about 40°C in ad...

Embodiment 2

[0061] 1. Preparation of liver fluke-specific IgG antibody functionalized gold nanorods

[0062] 1.1 Preparation and purification of gold nanorods

[0063] 1.1.1 Preparation of gold crystal seed solution

[0064] Firstly, gold nanoparticle seeds with small particle size are prepared. Mix cetyltrimethylammonium bromide solution (CTAB0.2mmol / L) 1.88mL, chloroauric acid solution (HAuCl 4 2mmol / L) 625μL, pure water 1.37mL and stir well. Take out the sodium borohydride (NaBH 40.01M) currently prepared in the 4°C refrigerator and add 450 μL to the above mixture, accelerate the stirring for several minutes until the color of the solution turns to tea red, continue to shake for 5 minutes, and then incubate in a 28°C incubator 2 hours.

[0065] 1.1.2 Growth solution preparation

[0066] Prepare several 50ml centrifuge tubes according to the needs of the experiment, add 11.875mL of CTAB (0.2M) that has been prepared and completely dissolved in each centrifuge tube, and then absorb ...

Embodiment 3

[0082] 1. Screening and identification of circulating antigen-related components

[0083] 1.1 SDS-PAGE analysis of circulating antigen-related components

[0084] 1.1.1 SDS-PAGE purification of excreted and secreted antigens

[0085] Add 1 / 4 volume of SDS-PAGE protein loading buffer (5×) to serum samples on days 3, 10, 17, and 31, and mix well. The mixture was heated in a boiling water bath for 5 to 10 minutes, and after heating, the mixture was centrifuged briefly and collected at the bottom of the tube. according to( Image 6 and Table 1) The protein concentration measured by the BCA protein quantification kit determines the loading amount and volume of the sample. And after electrophoresis treatment, staining and decolorization are carried out in sequence to obtain clearer protein bands and completely remove the staining background, then take pictures and record.

[0086] Finally, use the PAGE Gel Protein Micro Recovery Kit to elute the proteins in the recovered gel.

...

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Abstract

The invention discloses a clonorchis sinensis specific antibody functionalized nanogold rod. The nanogold rod is prepared by adopting a seed-mediated growth method, coating the gold nanorod with a sulfhydrylated clonorchis sinensis specific IgG antibody, and screening the clonorchis sinensis specific IgG antibody functionalized nano antibody by comparing the displacement of LSPR red peaks before and after coating. The invention further discloses the clonorchis sinensis specific antibody and application of the clonorchis sinensis specific antibody functionalized nanogold rod, and the clonorchissinensis specific antibody functionalized nanogold rod is specifically applied to detection of serum circulating antigen through the clonorchis sinensis specific antibody. According to the invention,the advantages of detection sensitivity and early stage are realized by utilizing a nanogold rod biotechnology, the operation is simple, the cost is low, and a new technical support is provided for early detection of clonorchis sinensis.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to the application of a nano-gold rod in the detection of serum circulating antigens by liver fluke-specific antibodies. Background technique [0002] Liver fluke, also known as Clonorchis sinensis, was first discovered in the bile duct of an overseas Chinese corpse in Calcutta, India in 1874. Because the testes of the parasite are branched, it is called Clonorchis sinensis. Adult worms mainly inhabit the hepatobiliary ducts of humans and carnivorous mammals such as cats and dogs, and occasionally also in the pancreatic ducts. The life cycle of liver fluke cannot be separated from the water environment. After entering the water, the eggs in the feces of the final host infected with liver fluke are swallowed by the first intermediate host, mainly snails, snails, and snails. A large number of cercariae are hatched and differentiated in the snail, and the mature cercari...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/6854G01N33/54346B22F9/24B82Y40/00B82Y30/00G01N2333/43547B22F1/054
Inventor 杨毅梅唐亮李延鹏杨晓燕肖文黄志旁廉晓丽吕艳
Owner DALI UNIV
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