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Detecting device for Brucella infection

A Brucella and detection device technology, applied in the field of detection devices for rapid detection of Brucella infection, can solve the problems of long detection cycle, poor specificity, strong interference, etc., meet the requirements of lower skill level, and low cost , Reduce the effect of missed diagnosis and misdiagnosis

Active Publication Date: 2016-10-12
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The technical problem to be solved by the present invention is to provide a detection device for Brucella infection, which can simultaneously detect Brucella antigens and antibodies in the sample, and the joint application of multiple outer membrane proteins can effectively reduce the risk of false positive results. It appears that the double-antigen sandwich and double-antibody sandwich methods can improve the sensitivity and specificity of detection compared with the traditional indirect method, and are used for the rapid detection of brucellosis; it solves the current detection cycle in the detection of brucellosis Long, poor specificity, low sensitivity and strong interference, prone to false positives or false negatives

Method used

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  • Detecting device for Brucella infection
  • Detecting device for Brucella infection
  • Detecting device for Brucella infection

Examples

Experimental program
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Effect test

Embodiment 1

[0023] 1. Preparation of nanoparticle-labeled detection antibody: Take 1mg of polystyrene fluorescent microspheres with a particle size of 110nm (time-resolved fluorescence, excitation: 360nm, emission: 615nm), wash the particles twice with PBS, and resuspend the particles with 1ml PBS. Add EDC and NHS to make the final concentrations 20mg / ml and 5mg / ml respectively; react at room temperature for 20min; centrifuge, discard the supernatant, and wash the particles twice with PBS; resuspend the particles, add 300μg Brucella lipopolysaccharide monoclonal antibody (C epitope); react at room temperature for 2 hours; add BSA to a final concentration of 1% (W / V), continue to react at room temperature for 2 hours; centrifuge, discard the supernatant, and wash the particles 3 times with PBS; use 10ml reconstitution solution (10mM Tris-HCl (pH8.0), 0.5% BSA) to resuspend the particles and set aside.

[0024] Preparation of nanoparticle-labeled OMP28 protein: take 1mg of polystyrene fluor...

Embodiment 2

[0033] 1. Preparation of nanoparticle-labeled detection antibody: Take 10ml of nano-gold particles with a particle size of 40nm, add 90μg Brucella lipopolysaccharide monoclonal antibody (C epitope); react at room temperature for 15min; add BSA to a final concentration of 1% (W / V), continue to react at room temperature for 15 minutes; centrifuge, resuspend the particles with 1ml reconstituted solution (10mM Tris-HCl (pH8.0), 0.5%BSA), and set aside.

[0034] 2. Preparation of nanoparticle-labeled OMP28 protein: Take 10ml of nano-gold particles with a particle size of 40nm, add 140μg of OMP28 protein (spliced ​​from 1-75AA and 135-225AA, pET-32a plasmid, expressed in E. coli BL21); react at room temperature for 15 minutes; Add BSA to a final concentration of 1% (W / V), and continue to react at room temperature for 15 minutes; centrifuge, and resuspend the particles with 1ml of reconstitution solution (10mM Tris-HCl (pH8.0), 0.5%BSA), and set aside.

[0035] 3. Preparation of chr...

Embodiment 3

[0043] 1. Preparation of nanoparticle-labeled detection antibody: Take 10ml of nano-gold particles with a particle size of 40nm, add 90μg Brucella lipopolysaccharide monoclonal antibody (C epitope); react at room temperature for 15min; add BSA to a final concentration of 1% (W / V), continue to react at room temperature for 15 minutes; centrifuge, resuspend the particles with 1ml reconstitution solution (10mM Tris-HCl (pH8.0), 0.5%BSA), and set aside.

[0044] 2. Preparation of nanoparticle-labeled OMP28 protein: Take 10ml of nano-gold particles with a particle size of 40nm, add 140μg of OMP28 protein (spliced ​​from 1-75AA and 135-225AA, pET-32a vector, expressed in Escherichia coli BL21); react at room temperature for 15 minutes; Add BSA to a final concentration of 1% (W / V), and continue to react at room temperature for 15 minutes; centrifuge, and resuspend the particles with 1ml of reconstitution solution (10mM Tris-HCl (pH8.0), 0.5%BSA), and set aside.

[0045] 3. Preparati...

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Abstract

The invention discloses a detecting device for Brucella infection. The detecting device comprises a test strip; a Brucella antigen detection area and a Brucella antibody detection area are arranged on a chromatographic membrane of the test strip. According to the detection device, a Brucella antigen and a Brucella antibody in a sample can be detected simultaneously; various outer membrane proteins are applied in a combined manner, so that the occurrence of false positive results can be reduced effectively; a double-antigen sandwich method and a double-antibody sandwich method are adopted, so that the sensitivity and the specificity of detection can be higher than those obtained according to the conventional indirect method, and the rapid detection of brucellosis is realized; the problems of high interference, and high false positive result or false negative result occurrence rate caused by long detection period, poor specificity and low sensitivity in the conventional brucellosis detection process are solved.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a detection device for rapid detection of Brucella infection. Background technique [0002] Brucellosis (brucellosis for short) is a zoonotic infectious disease that seriously threatens the life and health of humans and various animals. Sick sheep, cattle and other diseased animals are the main source of infection for brucellosis. Brucella can be transmitted through damaged skin and mucous membranes, digestive tract and respiratory tract. In the acute stage, the main manifestations are fever, fatigue, sweating, muscle and joint pain, and enlargement of liver, spleen and lymph nodes. Most cases in the chronic phase show joint damage. Brucellosis is a Class B infectious disease stipulated in my country's "Law on the Prevention and Control of Infectious Diseases". According to the 2012 statistical report of my country's CDC, the number of human brucellosis has increased to mor...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/558
CPCG01N33/558G01N33/56911G01N2400/50G01N2469/20
Inventor 黎诚耀李金峰张玲李文敏
Owner SOUTHERN MEDICAL UNIVERSITY
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