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Detection card for rapidly detecting nonstructural protein antibodies of FMDV (foot-and-mouth disease virus) in sera

A technology for foot-and-mouth disease virus and non-structural protein is applied in the field of detection cards for rapid detection of foot-and-mouth disease virus non-structural protein antibodies in serum. The effect of prevention and control ability and high sensitivity

Inactive Publication Date: 2018-04-03
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

ELISA operation is complex and labor-intensive; special laboratories and professionals are required to operate; the experiment takes a long time and the results are slow
Although DIBA and colloidal gold chromatography test strips are convenient to detect and do not require professional technicians and working environment; but their detection sensitivity is low, and they can only be qualitative, and the detection range is narrow

Method used

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  • Detection card for rapidly detecting nonstructural protein antibodies of FMDV (foot-and-mouth disease virus) in sera
  • Detection card for rapidly detecting nonstructural protein antibodies of FMDV (foot-and-mouth disease virus) in sera
  • Detection card for rapidly detecting nonstructural protein antibodies of FMDV (foot-and-mouth disease virus) in sera

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment one, the prokaryotic expression of 2C, 3AB and 3ABC protein of foot-and-mouth disease virus nonstructural protein

[0038] Using bioinformatics technology to analyze the main non-structural protein antigen genes 2C, 3AB and 3ABC of foot-and-mouth disease virus, design and synthesize the main non-structural protein antigen genes 2C, 3AB and 3ABC of foot-and-mouth disease virus, and insert the above genes into the prokaryotic expression plasmid pET-28a respectively (+), transform BL21(DE3) pLysS competent cells to screen positive recombinant bacteria. Select the positive recombinant clones and add them to 100ml LB culture solution containing kanamycin, shake culture at 220rpm at 37°C, when OD 600 When = 0.4-0.6, after adding IPTG with a final concentration of 0.6Mm, continue to culture for 4-6 hours, collect the culture by centrifugation, resuspend the precipitate with an appropriate amount of PBS, ultrasonically break it, and centrifuge at 10,000g for 30min. ...

Embodiment 2

[0039] Embodiment two, prepare the detection card of foot-and-mouth disease virus nonstructural protein antibody

[0040] Determine the cut-off value of nonstructural protein antibody of foot-and-mouth disease virus: use the positive serum standard (including strong positive, medium positive and weak positive) containing the nonstructural protein antibody of foot-and-mouth disease virus calibrator, and the negative standard of nonstructural protein antibody of foot-and-mouth disease virus each 2 share. Add the above-mentioned calibration solutions of different concentrations into the sample holes of the assembled test card, and after 15 minutes of chromatography, the test is carried out by a tomography scanner, and the test results obtained 8 times are processed by the client, and the client calculates The fluorescence signal intensity values ​​of the detection line and quality control line corresponding to the standard product, and according to this data, carry out statistica...

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Abstract

The invention discloses a detection card for rapidly detecting nonstructural protein antibodies of an FMDV (foot-and-mouth disease virus) in sera. The detection card comprises a detection card housingand a test strip assembled in a detection card housing, wherein the test strip comprises a plastic base plate with a pressure-sensitive adhesive, and a sample pad, a marker pad, a nitrocellulose membrane and absorbent paper are adhered to the base plate in sequence; the marker pad comprises a carrier substrate and a marker, and is a membrane formed by spraying lanthanide fluorescence detection microspheres and lanthanide fluorescence quality control microspheres on the carrier substrate; the nitrocellulose membrane is coated with recombinant protein of FMDV nonstructural protein as a detection line and coated with rabbit anti-chicken lgY antibodies as a control line, and the markers are the fluorescence detection microspheres labeled with FMDV nonstructural protein recombinant antigens and the fluorescence quality control microspheres labeled with chicken lgY antibodies. The detection card can realize rapid detection of the nonstructural protein antibodies of the FMDV on site and in the field and is used for distinguishing virus infection animals from vaccinated animals.

Description

technical field [0001] The invention relates to a detection card and its use, in particular to a detection card for rapid detection of non-structural protein antibodies of foot-and-mouth disease virus in blood serum. Background technique [0002] Foot-and-Mouth Disease (FMD) is a major animal disease caused by the Foot-and-Mouth Disease Virus (FMDV), which is mainly economical species of pigs, cattle, and sheep. The disease has many transmission routes, fast speed, wide endemic area, and wide host spectrum. Once the disease occurs, it will not only cause huge economic losses, but also cause bad political impact. It has attracted great attention from all countries in the world and is known as "political and economic disease". . , the World Organization for Animal Health (OfficeInternational des Epizooties, OIE) lists it as an animal disease that must be notified, and my country lists it as the first of the Class A animal infectious diseases. With the acceleration of the glo...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569C12N15/70C12N15/42
CPCC07K14/005C12N15/70C12N2770/32122G01N33/56983G01N33/68G01N2333/09
Inventor 常惠芸邵军军吴俊清章健刘择众吴冠英李扬帆王泽洲赵付荣张永光
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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