Test paper card for detecting swine transmissible gastroenteritis virus antibody and preparation and detection method of test paper card

A technology for infectious and gastroenteritis, applied in the field of immunological detection, can solve the problems of large error, low sensitivity, poor stability, etc., and achieve the effect of shortening time, high sensitivity and good stability

Inactive Publication Date: 2018-04-20
洛阳现代生物技术研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the colloidal gold technology has the advantages of fast, convenient, and no need for special instruments, it also has disadvantages: the colloidal gold labeling method is electrostatically adsorbed, and the stability is poor; the results of the colloidal gold technology are judged by naked eyes, with large errors and low sensitivity.

Method used

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  • Test paper card for detecting swine transmissible gastroenteritis virus antibody and preparation and detection method of test paper card
  • Test paper card for detecting swine transmissible gastroenteritis virus antibody and preparation and detection method of test paper card
  • Test paper card for detecting swine transmissible gastroenteritis virus antibody and preparation and detection method of test paper card

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1, the preparation of test paper card

[0025] 1. Preparation of fluorescent microsphere-antibody complexes

[0026] (1) Cleaning: Take 50μl of fluorescent microspheres into a 1.5ml centrifuge tube, add 1ml of 0.01M MES buffer, shake and mix, centrifuge at 15000r / min for 15min, discard the supernatant, add 1ml of 0.01M MES buffer, and ultrasonically beat 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;

[0027] (2) Activation: add 250μl EDC solution to 1ml of microsphere solution, activate in the dark for 2h, centrifuge at 15000r / min for 15min, discard the supernatant, add 1ml of 0.01M MES buffer, ultrasonically break up the microspheres, and centrifuge at 15000r / min 15min, discard the supernatant;

[0028](3) Labeling: add 1ml of 0.05M borate buffer, disperse by ultrasonic, add 20μg anti-E2 protein monoclonal antibody and mix well, an...

Embodiment 2

[0048] Embodiment 2, detection method and comparative test of porcine transmissible gastroenteritis virus antibody in pig anal swab

[0049] 1. Wet the cotton swab with physiological saline and wipe the pig feces and rectum. Immediately insert the cotton swab into the sample tube containing the diluent, fully stir the cotton swab until all the samples on the cotton swab are dissolved in the diluent, and squeeze the cotton swab on the tube wall. Discard after drying, and the sample solution is left to stand for later use; the diluent is PBS with pH 7.4 and 0.05mol / L;

[0050] 2. 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;

[0051] 3. Take the prepared clarified sample supernatant with the matching straw, and drop 2-3 drops (about 60 μl) vertically and slowly into the sample well;

[0052] 4. Read the result in 5-10 minutes, insert the test paper card into the card slot of the fluorescence immunoass...

Embodiment 3

[0056] Embodiment 3, colloidal gold-labeled anti-E2 protein monoclonal antibody and the comparison test of the stability of fluorescent microsphere-labeled anti-E2 protein monoclonal antibody

[0057] 1. Place the prepared colloidal gold-antibody complex binding pad and the fluorescent microsphere-antibody complex binding pad in a blast drying oven at 37°C;

[0058] 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 samples and positive samples respectively, and observe their stability;

[0059] 3. The results are shown in Table 1 below.

[0060]

[0061] It can be seen from the above table 1 that the detection effect of the colloidal gold-labeled antibody complex decreases with the prolongation of the destruction test time, and the detection situation changes significantly at the 20th week; while the test strip using the fluorescent microsphere-antibody complex binding pad The positive samples showe...

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Abstract

The invention relates to a test paper card for detecting porcine transmissible gastroenteritis virus antibody, preparation and detection method thereof, and belongs to the field of immunological detection. The test paper card includes a jammed case and a test strip, and the test strip includes a bottom plate and sequentially lapped and pasted on the Absorbent pad, detection pad, binding pad and sample pad on the base plate; the detection pad is a nitrocellulose membrane with a quality control line C and a detection line T, the quality control line C is coated with goat anti-mouse monoclonal antibody, and the detection line T is Coated with inactivated porcine transmissible gastroenteritis virus; the binding pad is a glass cellulose membrane embedded with time-resolved fluorescent microsphere-labeled anti-E2 protein monoclonal antibody; the sample pad is dried glass after soaking in the sample treatment solution Cellulose film. The test paper card prepared by the invention has better stability and higher sensitivity, and can achieve the purpose of semi-quantitative detection through fluorescence signal analysis.

Description

technical field [0001] The invention belongs to the field of immunological detection, and in particular relates to a test paper card for detecting porcine transmissible gastroenteritis virus antibody, its preparation and its detection method. Background technique [0002] Porcine transmissible gastroenteritis (TGE) is piglet gastroenteritis caused by porcine transmissible gastroenteritis virus (TGEV). Viral infectious disease characterized by high mortality. TGEV is a member of group 1 of the genus αcoronavirus of the family Coronaviridae and subfamily Coronaviridae. TGE mostly occurs in winter and spring, and occurs in all regions of the world. It is endemic or occurs in local areas. The virus can be transmitted through direct contact, milk, and respiratory tract. The recovery time of pigs can be as long as 8 weeks. The main source of infection in pig farms. The disease is usually co-infected with Escherichia coli and rotavirus, leading to increased mortality of suckling...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531
CPCG01N33/577G01N33/531
Inventor 李秀梅闫玉良李俊俊杨丹刘近张晓亮李玉婉石方方王芳
Owner 洛阳现代生物技术研究院有限公司
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