Supercharge Your Innovation With Domain-Expert AI Agents!

Test paper applied to detection of coxiella burnetii antibody, preparation method of test paper and kit

A technology for detecting Coxie bodies and test strips, which is applied to measuring devices, instruments, scientific instruments, etc., can solve the problems of low effective antigen content, reduced sensitivity, and reduced amount of labeled antibodies, so as to achieve accurate, reliable and safe detection results Sexual problems improve, amplify the effect of the response effect

Active Publication Date: 2015-02-04
CHINESE ACAD OF INSPECTION & QUARANTINE
View PDF4 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] At present, the detection technologies for Coxiella burnetii (commonly known as Q-fever Rickettsia) antibodies are mainly immunological methods such as ELISA, chemiluminescence, and colloidal gold. The detection of absorbance by wind and light photometry is required, and it takes a long time; the fluorescent reagents required in the chemiluminescence method are relatively expensive, and the detection requires a specific fluorescence analysis instrument; the colloidal gold detection method is suitable for on-site detection, but the detection sensitivity is limited. Since the colloidal gold is very unstable after connecting the antibody to be labeled or the protein to be labeled, it is easy to detach the antibody due to the change of charge, resulting in a greatly reduced amount of labeled antibody per colloidal gold, resulting in a decrease in sensitivity
Moreover, the preparation of the colloidal gold reagent uses chloroauric acid, and the cost of the reagent is relatively high, resulting in a relatively high overall cost of the immunochromatography test paper.
[0003] One of the most critical factors in immunological detection methods such as ELISA, chemiluminescence, and colloidal gold is the selection of antigens and antibodies. If the antigen is prepared directly from the culture medium of Coxiella basil, there are complex antigen components, low content of effective antigen, and miscellaneous proteins. Many, poor immunogenicity, and high biosafety risks, and when used in immunological testing, it is easy to lead to positive results

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Test paper applied to detection of coxiella burnetii antibody, preparation method of test paper and kit
  • Test paper applied to detection of coxiella burnetii antibody, preparation method of test paper and kit
  • Test paper applied to detection of coxiella burnetii antibody, preparation method of test paper and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Preparation of Coxiella bezieri antigen

[0048] Using recombinant proteins to prepare antigens of corresponding pathogens is one of the ways to effectively solve the safety problem of pathogens and improve the detection rate of pathogens. At present, there is no report of a recombinant protein that can effectively serve as an antigen of Coxiella bezieri.

[0049] The specific steps for preparing the Coxiella bezieri antigen are as follows:

[0050] a. Construction of recombinant plasmids

[0051] For the selection of the Coxiella beinii antigen, the inventors of the present invention conducted multiple tests, and finally selected the Coxiella beinii envelope P1 protein, whose amino acid sequence reference (GenBank sequence number: AY249911.1) was based on Escherichia coli Escherichia coli O127:H6 preferred codons were used to modify the gene sequence, and the secondary structure of the recombinant protein was screened through bioinformatics (see the referen...

Embodiment 2

[0058] Example 2 Preparation of test strips for the detection of Coxiella basilica antibodies

[0059] A. SPA labeling magnetic nanoparticles, and preparing magnetic nanoparticle carrier pads:

[0060] Take 100 μl ferroferric oxide magnetic nanoparticle solution (the particle size of ferric oxide tetroxide is preferably 12nm), add 700 μl water, 200 μl 25% glutaraldehyde solution, shake for 3 h; wash 1-2 times with 1 ml water, and wash 2 times in PBS The second time is to carry out carboxylation treatment on ferric ferric oxide magnetic nanoparticles; add 500 μl of 2 mg / ml SPA, shake for 3 hours, and the pH value is 7.6; add 500 μl of 0.5% BSA, shake for 30 minutes, and block; add 0.01% spit Warm-20 PBS with a concentration of 0.01M, wash 3-4 times; add 1mL resuspension solution, wherein the resuspension solution contains 0.1% Tris base, 1% BSA and 5% sucrose by weight percentage, mix well, Spray on glass fiber membrane and dry at 37°C for 4 hours;

[0061] B. Coated nitrocel...

Embodiment 3

[0066]Example 3 The detection test paper of Coxella basidiobacillus antibody prepared by the present invention and the detection of the sensitivity of colloidal gold immunochromatography test paper

[0067] Wherein, see embodiment 2 for the preparation method of the detection test paper of the Coxiella bezii antibody prepared by the present invention;

[0068] The preparation method of colloidal gold immunochromatography test paper is as follows:

[0069] Colloidal gold immunochromatography test paper was prepared by conventional methods, wherein SPA-labeled colloidal gold particles were used, the detection zone on the nitrocellulose membrane was coated with Cox's body recombinant antigen, and the quality control zone was coated with goat anti-rabbit IgG.

[0070] The test strips prepared in Example 2 were used to detect the Coxiella bezieri antibody serum respectively, diluted with fetal bovine serum to 1:10, 1:100, 1:1000, 1:10000, 1:100000, 1:1000000, At the same time, tak...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Particle sizeaaaaaaaaaa
Login to View More

Abstract

The invention discloses test paper applied to detection of a coxiella burnetii antibody. The test paper comprises a sample pad, a Fe3O4 magnetic nano-particle carrier pad, a nitrocellulose membrane and a water absorption pad which are sequentially superposed and attached to a substrate card, wherein the Fe3O4 magnetic nano-particle carrier pad is a glass fiber membrane fixed with a staphylococal protein A (SPA) marked magnetic nano-particles; the nitrocellulose membrane is provided with a detection zone and a quality control zone; the detection zone is covered by a coxiella burnetii antigen; the quality control zone is covered by an antibody which can be combined with SPA. The kit applied to detection of the coxiella burnetii antibody comprises the detection test paper and a peroxidase substrate color-developing solution. The kit is capable of effectively detecting the coxiella burnetii antibody, and is stable and reliable in detection result, high in specificity, high in sensitivity and simple to operate; the sensitivity of the test paper is over ten times of that of the ordinary colloidal gold test paper; the detection result can be obtained by naked eyes without detection instrument.

Description

technical field [0001] The invention relates to the field of biological detection reagents, in particular to a detection test paper for Coxiella bezieri antibody, a preparation method for the test paper and a kit. Background technique [0002] At present, the detection technologies for Coxiella burnetii (commonly known as Q-fever Rickettsia) antibodies are mainly immunological methods such as ELISA, chemiluminescence, and colloidal gold. The detection of absorbance by wind and light photometry is required, and it takes a long time; the fluorescent reagents required in the chemiluminescence method are relatively expensive, and the detection requires a specific fluorescence analysis instrument; the colloidal gold detection method is suitable for on-site detection, but the detection sensitivity is limited. Since the colloidal gold is very unstable after being connected with the antibody to be labeled or the protein to be labeled, it is easy to cause the antibody to detach due t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N33/558
CPCG01N33/558
Inventor 杨宇韩辉马爱敏赵婷婷王静徐宝梁
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com