Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Immunity test strip based on fluorescent microsphere for detecting infectious pleural pneumonia antibody of goat as well as preparation and detection method

A fluorescent microsphere, pleuropneumonia technology, applied in the field of immunological detection, can solve the problems of low sensitivity, poor repeatability or specificity, and need laboratory instruments, and achieves the effect of high sensitivity and shortened time.

Inactive Publication Date: 2018-03-02
LUOYANG LAIPSON INFORMATION TECH +1
View PDF9 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Rapid and accurate diagnosis is beneficial to the prevention and control of Mccp. At present, there are many methods for the detection of Mccp antibodies that have been reported at home and abroad. The main techniques include indirect hemagglutination test, direct and indirect fluorescent antibody test, complement fixation test, and enzyme-linked immunosorbent immunoassay. Adsorption test (ELISA) and PCR technology recommended by OIE, but there are many disadvantages such as complex operation, poor repeatability or specificity, and the need for laboratory equipment, etc., especially not suitable for rapid, easy-to-operate and grass-roots use in the actual diagnosis of animal diseases in my country requirements
In veterinary clinical disease diagnosis and inspection and quarantine, it is usually used in farms and grassroots units. Faced with the needs of large-scale samples, large-scale censuses and on-site diagnosis, immune colloidal gold technology is the most widely used in the market. It has The detection is fast, convenient, and does not require special instruments, but there are also disadvantages: the colloidal gold labeling method is electrostatic adsorption, which has poor stability; the colloidal gold technical results are judged by naked eyes, with large errors and low sensitivity; it cannot be used for quantitative detection

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
  • Immunity test strip based on fluorescent microsphere for detecting infectious pleural pneumonia antibody of goat as well as preparation and detection method
  • Immunity test strip based on fluorescent microsphere for detecting infectious pleural pneumonia antibody of goat as well as preparation and detection method
  • Immunity test strip based on fluorescent microsphere for detecting infectious pleural pneumonia antibody of goat as well as preparation and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1, the preparation of test paper card

[0026] 1. Preparation of binding pads

[0027] (1) Cleaning: Take 50 μL of fluorescent microspheres into a 1.5 mL centrifuge tube, add 1 mL of 0.01M MES buffer, shake and mix, centrifuge at 15,000 r / min for 15 min, discard the supernatant, add 1 mL of 0.01M MES buffer, and ultrasonically Loose microspheres; repeat this step three times, the purpose of cleaning the microspheres has been achieved, and finally the microsphere liquid after cleaning is obtained;

[0028] (2) Activation: Add 250 μL of EDC solution to 1 mL of microsphere solution after washing, activate in the dark for 2 h, centrifuge at 15,000 r / min for 15 min, discard the supernatant, add 1 mL of 0.01M MES buffer, and disperse the microspheres by ultrasonication, 15,000 r / min Centrifuge for 15min, discard the supernatant;

[0029] (3) Labeling: Add 1 mL of 0.05M borate buffer, disperse by ultrasonication, add 20 μL of Mycoplasma goatum recombinant protein...

Embodiment 2

[0048] Embodiment 2, detection method and comparison test of goat infectious pleuropneumonia antibody in goat serum

[0049] 1. Tear open the aluminum foil packaging bag of the test card, take out the test card, and place it on a flat and clean table;

[0050] 2. Absorb freshly collected goat serum with a matching straw, and drop 2-3 drops (about 60 μl) vertically and slowly into the sample well;

[0051] 3. Read the result in 5-10 minutes, insert the test paper card into the card slot of the fluorescence immunoassay analyzer in the direction of the arrow shown on the card cover, and read the T / C value;

[0052] 4. Judgment criteria: T / C value < 0.1 is negative; T / C value ≧ 0.1 is positive; C value is 0, invalid;

[0053] 5. Comparison with colloidal gold test strips: select 20 weakly positive samples detected by goat infectious pleuropneumonia hemagglutination diagnostic kit, test the results with colloidal gold test strips, and detect 5 positive and 15 negative; Fluorescen...

Embodiment 3

[0055] Example 3. Comparison test of the stability of colloidal gold-labeled Mycoplasma pneumoniae recombinant protein and fluorescent microsphere-labeled recombinant protein against Mycoplasma pneumoniae

[0056] 1. Place the prepared colloidal gold-antigen complex binding pad and the fluorescent microsphere-antigen complex binding pad in a 37°C blast drying oven;

[0057] 2. On the 3rd day, 5th day, 1 week, 2 weeks, 4 weeks, 8 weeks, 12 weeks, 20 weeks, 36 weeks, and 48 weeks, test the negative serum and positive serum respectively, and observe their stability;

[0058] 3. The result is as follows

[0059]

[0060] As shown in the above table, the detection effect of the colloidal gold-labeled antigen complex decreased with the extension of the destruction test time, and the detection situation changed significantly at the 20th week; while the test strip using the fluorescent microsphere-antigen complex binding pad The positive sera showed a downward trend in the 36th we...

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
Diameteraaaaaaaaaa
Login to View More

Abstract

The invention relates to an immunity test strip based on a fluorescent microsphere for detecting an infectious pleural pneumonia antibody of a goat as well as a preparation and detection method, whichbelong to the detection field of immunity. The test strip comprises a shell and a test strip, wherein the test strip comprises a bottom plate as well as a water absorption pad, a detection pad, a combination pad and a sample pad which are successively lapped and adhered onto the bottom plate; a nitrocellulose membrane is provided with a quality control line C and a detection line T, the quality control line C covers a His tag monoclonal antibody, and the detection line T covers a goat mycoplasma pneumonia recombinant protein; the combination pad is a glass cellulose membrane coating with thegoat mycoplasma pneumonia recombinant protein marked by time resolution fluorescent microspheres; and the sample pad is a dry glass cellulose membrane after being soaked in sample treatment liquid. The test strip prepared by the invention has better stability and higher sensitivity, and can achieve a purpose of rapid semi-quantitative detection by virtue of the fluorescent signal analysis.

Description

technical field [0001] The invention belongs to the field of immunological detection, and in particular relates to an immune test strip for detecting goat infectious pleuropneumonia antibody based on fluorescent microspheres, a preparation method and a detection method. Background technique [0002] Goat contagious pleuropneumonia (Mccp), also known as Mycoplasma capricum pneumonia, is the pathogen of Mycoplasma capricosum pneumonia, which belongs to the subspecies of Mycoplasma capricosum, and is a unique acute or chronic highly contagious infectious disease of goats. The disease is clinically characterized by high fever, cough, serous and fibrinous inflammation in the lungs and pleura, and has a high mortality rate. In recent years, the pathogens of Mccp have been isolated or detected in Gansu, Shandong, Qinghai and other places in China, and its morbidity and mortality are on the rise, which has caused great economic losses to the goat breeding industry in my country. ...

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): G01N33/533G01N33/558G01N33/543G01N33/577
CPCG01N33/533G01N33/54313G01N33/558G01N33/577
Inventor 李秀梅陈小云任雪建张志鹏王俊峰张小辉王蓝天
Owner LUOYANG LAIPSON INFORMATION TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com