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Colloidal gold immunochromatography test paper for detecting canine Echinococcus granulosus infection, and production method thereof

A technology of Echinococcus granulosus and immunochromatographic test paper, which is applied in the field of immunology to achieve the effect of improving sensitivity, ensuring safety and high specificity

Inactive Publication Date: 2016-11-09
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the development of colloidal gold test strips for the detection of Echinococcus granulosus infection by using the monoclonal antibody prepared by EdiagA864 and the rabbit polyclonal antibody

Method used

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  • Colloidal gold immunochromatography test paper for detecting canine Echinococcus granulosus infection, and production method thereof
  • Colloidal gold immunochromatography test paper for detecting canine Echinococcus granulosus infection, and production method thereof
  • Colloidal gold immunochromatography test paper for detecting canine Echinococcus granulosus infection, and production method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Preparation of colloidal gold test strips for detection of Echinococcus granulosus infection in dogs

[0048] (1) Firing of colloidal gold

[0049] Using trisodium citrate reduction method, to 99mL ddH 2 Add 1 mL of 1% chloroauric acid to O, and after boiling, add 1 mL of 1% trisodium citrate to prepare 40 nm colloidal gold particles.

[0050] (2) Determination of the optimal pH of the gold-labeled antibody

[0051] Use K 2 CO 3 Gradient labeling method, by observing the color change, determine the optimal pH of the gold standard.

[0052] (3) Preparation of gold-labeled antibody

[0053] Best K 2 CO 3 The amount of addition is to label the monoclonal antibody with colloidal gold, and the labeled gold-labeled antibody is dissolved in the reconstitution solution and stored at 4°C for later use or directly dropped on the gold-labeled pad.

[0054] (4) Determination of T-line polyclonal antibody coating concentration on NC membrane

[0055] Dilute the T-line polyc...

Embodiment 2

[0070] Colloidal gold immunochromatographic test strip performance test

[0071] In order to test the test performance of the test strip, a series of tests are required, including specificity tests, sensitivity tests, repeatability tests, and shelf life tests.

[0072] (1) Specificity test

[0073] Take Echinococcus granulosus reference-positive feces, Ascaris canis-positive feces, Giardia-positive feces, and healthy dog ​​feces as negative controls, and operate according to conventional methods to judge whether there is cross-reaction between various feces, so as to evaluate Specificity of test strips (see Figure 4 ).

[0074] (2) Sensitivity test

[0075] Doubling dilution (1:1, 1:2, 1:4, 1:8, 1:16) of the positive Echinococcus canis feces, and observe the color development of the test strip according to the routine operation. When the T line cannot be observed with the naked eye, and the C line is still clearly visible, the dilution at this time is the minimum detectio...

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Abstract

The invention discloses a production method of colloidal gold immunochromatography test paper strip for detecting canine Echinococcus granulosus infection. The production method comprises the following steps: cloning, expressing and purifying an EdiagA864 recombinant protein, preparing and purifying a monoclonal antibody and a rabbit polyclonal antibody, sintering a colloidal gold solution by using a trisodium citrate reducing technology, labeling the monoclonal antibody with colloidal gold to produce a colloidal gold pad, and assembling the test paper strip, and using Echinococcus granulosus EdiagA864 polyclonal antibody as adetection line and a goat anti-mouse antibody as a quality inspection line. The test paper strip has the characteristics of fastness and convenience in operation, no special devices, clear detection result, rapid judgment, and suitableness for clinic and onsite rapid diagnosis and large-scale epidemiology investigation of pastoral areas. The test paper strip has high specificity and sensitivity; and the method uses Echinococcus granulosus EdiagA864 monoclonal antibody and polyclonal antibody sandwich, the monoclonal antibody guarantees the specificity of the method, and the polyclonal antibody improves the sensitivity of the method. Compared with traditional arecoline catharsis methods, the test paper strip has the advantages of guaranteeing of the safety of detection personnel, and avoiding of egg infection.

Description

technical field [0001] The invention relates to a colloidal gold immunochromatographic test paper for detecting Echinococcus granulosus infection, which is used for feces detection of Echinococcus granulosus infection and belongs to the technical field of immunology. Background technique [0002] Canine Echinococcus granulosus is a zoonotic parasitic disease caused by Echinococcus granulosus that causes mechanical damage to the internal organs of the host. The disease is prevalent worldwide, seriously threatening the health of humans and animals, and hindering the development of animal husbandry. Dogs are an important terminal host of the disease and an important part of the epidemic. At present, there are many diagnostic methods for Echinococcus granulosus, among which OIE regards autopsy and arecoline catharsis as the "gold standard" for the detection of Echinococcus granulosus in dogs. However, the "gold standard" method has many disadvantages such as complex operation,...

Claims

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Application Information

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IPC IPC(8): G01N33/558
CPCG01N33/558
Inventor 宫鹏涛张西臣高珺珊李建华张壮志杨举李赫李棕松杨正涛尹继刚
Owner JILIN UNIV
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