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Colloid selenium test paper for detecting pulpy kidney, and type of pathogenesis bacteria

A technology for Clostridium welchii and detection test paper, which is applied in the field of selenium gel detection test paper to achieve the effects of stable reagents, good specificity, and firm combination

Inactive Publication Date: 2014-01-29
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there is no diagnostic test paper for Clostridium welchii disease based on colloidal selenium chromatography at home and abroad, including literature reports and invention patents.

Method used

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  • Colloid selenium test paper for detecting pulpy kidney, and type of pathogenesis bacteria

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Experimental program
Comparison scheme
Effect test

Embodiment

[0017] Embodiment (one) extraction of antigen

[0018] The extraction of the antigen Clostridium welchii α-toxin went through two stages of crude extraction and fine extraction.

[0019] Crude extraction: Inoculate Clostridium welchii type A strains in semi-solid blister medium, culture at 37°C for 10 hours, inoculate 0.5% in semi-solid blister medium, culture at 37°C for 6 hours, and sterilize with EKS filter plate Filtration, the filtrate is crude alpha toxin.

[0020] Fine extraction: Crude α-toxin (culture filtrate) was salted out with 70% saturated ammonium sulfate for 30 minutes, centrifuged at 10,000 rpm for 15 minutes, and the supernatant was discarded; the precipitate was suspended with an equal amount of normal saline solution, salted out with 20% saturated ammonium sulfate for 30 minutes, 10,000 rpm Centrifuge for 15 minutes, then salt out the supernatant with 60% saturated ammonium sulfate for 30 minutes, centrifuge at 10,000 rpm for 15 minutes, discard the supern...

Embodiment ( 4

[0070] Embodiment (four) selenium mark monoclonal antibody

[0071] The process of selenium-labeled Clostridium welchii alpha toxin monoclonal antibody 1 is as follows:

[0072] 1. Preparation of Clostridium welchii α-toxin monoclonal antibody 1 solution to be labeled

[0073] Dialyze Clostridium welchii α-toxin monoclonal antibody 1 in 0.005mol / L pH7.0NaCl solution at 4°C overnight to remove excess salt ions, then centrifuge at 3500rpm at 4°C for 1h to remove polymers, and buffer with 0.01 PBS for precipitation Solution (pH8.0) was diluted to 1mg / ml for later use.

[0074] 2. Preparation of colloidal selenium solution to be labeled

[0075] First take a certain amount of glucosidan with a concentration of 0.800‰ and a concentration of 1.00×10 -3 mol / L H 2 SeO 3 Put the solution in a 10mL colorimetric tube, mix it evenly and let it stand for a while, add a certain amount of 5.00×10 -3 mol / L ascorbic acid (Vc) solution, added H2 SeO 3 The ratio of the amount of substance...

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Abstract

A disease germ type of colloid Se test paper for testing Clostridium welchii disease consists of plastic base plate, Se labeled pad formed by glass cellulose film with detection region and control region, sampling-adding film, water absorption paper and reaction film formed by nitric acid cellulose film with detection region and control region.

Description

1. Technical field [0001] The invention relates to a selenium gel detection test paper for Clostridium welchii disease and its pathogenic bacteria type, in particular to a test paper capable of rapidly detecting Clostridium welchii α toxin in samples and identifying the pathogenic bacteria of Clostridium welchii disease The type of test paper can be used in veterinary clinics, animal husbandry, food safety, public health, water quality monitoring and related fields. 2. Background technology [0002] Clostridium Welchii, also known as Clostridium perfringens, is widely distributed in nature, is a typical opportunistic pathogen, and is also a member of the normal intestinal flora of animals. It can cause various animal necrotic enteritis, enterotoxemia, traumatic gangrene in humans and animals, and human food poisoning. Clostridium welchii can produce a variety of exotoxins, among which α, β, ε and ι are the main lethal toxins. According to the neutralization test of the mai...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/569G01N33/558G01N33/544
Inventor 李红玉晏磊何丽娟陈瑜
Owner LANZHOU UNIVERSITY
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