Making method of semi-quantitative tetrodotoxin detection card

A technology of tetrodotoxin and its production method, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of too specialized test strips, high cost of monoclonal antibodies, cumbersome instrument detection, etc. Small size effect

Inactive Publication Date: 2014-11-12
FISHERIES RES INST OF FUJIAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] The invention provides a method for making a semi-quantitative tetrodotoxin detection card, one of the purposes of which is to overcome the defects of the existing technology that the instrument detection is relatively cumbersome and too...

Method used

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  • Making method of semi-quantitative tetrodotoxin detection card
  • Making method of semi-quantitative tetrodotoxin detection card

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A semi-quantitative tetrodotoxin test card, such as figure 1 and figure 2 As shown in , it includes a carrier pad 7, a nitrocellulose membrane 4, a colloidal gold sample combination pad 1, an absorbent pad 6, an adhesive tape layer 2 and a cassette case.

[0037] The production process of the test card is divided into two steps, that is, the pretreatment of the early preparation and the later assembly. The pretreatment is mainly divided into the preparation of the colloidal gold sample combination pad and the processing of the chromatographic membrane. The specific steps are as follows:

[0038] 1) Preparation of colloidal gold sample combination pad 1: immunize mice with tetrodotoxin artificial antigen, take splenocytes from immunized mice and myeloma cells for fusion culture, screen positive hybridoma cells for in vitro culture, and clone them for expansion and culture and freeze them. BALB / c mice or their parental mice, choose BALB / c mice or their parental mice, fi...

Embodiment 2

[0045] The difference between embodiment 2 and embodiment 1 is:

[0046] 1) Preparation of colloidal gold sample combination pad 1: immunize mice with tetrodotoxin artificial antigen, take splenocytes from immunized mice and myeloma cells for fusion culture, screen positive hybridoma cells for in vitro culture, and clone them for expansion and culture and freeze them. BALB / c mice or their parental mice were selected, BALB / c mice or their parental mice were selected, and liquid paraffin was used for intraperitoneal injection of mice, and hybridoma cells were inoculated into the peritoneal cavity of mice 8 days later, and then 12 days later Collect the ascites, and then add 10 times the colloidal gold labeling solution to the ascites under the environment of pH11 for labeling. In order to avoid the formation of coagulated substances at the front end of the sample chromatography due to too many impurities in the ascites, prevent sample chromatography and combine with nitrocellulos...

Embodiment 3

[0050] The difference between embodiment 3 and embodiment 1 is:

[0051] 1) Preparation of colloidal gold sample combination mat: mice were immunized with tetrodotoxin artificial antigen, splenocytes from immunized mice were fused with myeloma cells for fusion culture, positive hybridoma cells were screened and cultured in vitro, clones were expanded and cultured and frozen, and BALB was selected / c mice or their parental mice, BALB / c mice or their parental mice were selected, and liquid paraffin was used for intraperitoneal injection of mice, and hybridoma cells were inoculated into the peritoneal cavity of mice 7 days later, and then collected 10 days later Ascites, and then add 15 times of colloidal gold labeling solution to the ascites under the environment of pH9 for labeling. In order to avoid the formation of coagulated substances at the front end of the sample chromatography due to too many impurities in the ascites, which prevents the sample from being chromatographed ...

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Abstract

The invention discloses a making method of a semi-quantitative tetrodotoxin detection card. The method comprises the following steps: 1, preparing a colloidal gold sample combination pad; 2, chromatographic membrane processing: uniformly drawing a tetrodotoxin artificial antigen forming a detection line and a quality control antibody forming a quality control line in the middle of a chromatographic membrane by using a membrane drawing instrument; and 3, assembling: splicing a carrier pad, the chromatographic membrane, the colloidal gold sample combination pad and a water absorption pad to form a layered structure, wherein the carrier pad is positioned at the bottom, the chromatographic membrane is arranged on the carrier pad, and the colloidal gold sample combination pad and the water absorption pad are respectively placed at two ends of the chromatographic membrane. The above obtained product allows tetrodotoxin to be rapidly semi-quantitatively analyzed through ocular estimation, and has the characteristics of small size, light weight, easy preservation and transportation, and realization of detection of a detected sample without separation or purification.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a method for preparing a semi-quantitative tetrodotoxin detection test paper. Background technique [0002] Since the 1990s, my country has reported 50 cases of snail poisoning, 369 people were poisoned, 43 died, and the poisoning mortality rate was 11.2%. The incidents of poisoning and death of snails occurred. In addition to textured snails, there are also cases of puffer fish poisoning every year. [0003] The main components of toxins contained in snails and puffer fish are tetrodotoxin and its derivatives. [0004] Tetrodotoxin is a non-protein, low-molecular-weight neurotoxin that can bind to sodium ion channels, thereby affecting the muscle and nerve functions of animals. Tetrodotoxin was first discovered in puffer fish and is widely distributed in terrestrial and aquatic organisms, such as dinoflagellate cysts, calcareous algae, arthropods, echinoderms, molluscs, wor...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/532
CPCG01N33/558G01N33/543
Inventor 苏捷姜琳琳吴靖娜张农刘智禹
Owner FISHERIES RES INST OF FUJIAN
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