Quantitative ELISA (Enzyme Linked Immunosorbent Assay) detection method for antigen 146S of foot and mouth disease A as well as kit and application thereof

A technology of detection kits and detection methods, applied in measurement devices, instruments, scientific instruments, etc., can solve problems such as being unsuitable for the detection of a large number of samples, complicated operations, and expensive instruments, and achieve an optimized vaccine production process, simple operation, and stability. Good results

Active Publication Date: 2013-05-08
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method also has its disadvantages: complex operation, expensive equipment, poor repeatability, not suitable for the detection of a large number of samples, and cannot distinguish serotypes
According to reports, the detection results of this method are in good agreement with the sucrose density gradient centrifugation method, and the operation is simpler than the latter; but like the latter, it cannot distinguish serotypes

Method used

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  • Quantitative ELISA (Enzyme Linked Immunosorbent Assay) detection method for antigen 146S of foot and mouth disease A as well as kit and application thereof
  • Quantitative ELISA (Enzyme Linked Immunosorbent Assay) detection method for antigen 146S of foot and mouth disease A as well as kit and application thereof
  • Quantitative ELISA (Enzyme Linked Immunosorbent Assay) detection method for antigen 146S of foot and mouth disease A as well as kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] A type A foot-and-mouth disease 146S antigen quantitative ELISA detection method comprises the following steps:

[0058] 1) Reserve type A foot-and-mouth disease standard control antigen:

[0059] For the type A foot-and-mouth disease standard control antigen, one part is 5 mL / bottle, stored at -70°C (thawed only once, for ELISA test); the other part is 3×100 mL, stored at -70°C (thawed only once, used Purification and quantification of 146S antigen (three times));

[0060] 2) Purify and quantify type A foot-and-mouth disease standard control antigen 146S:

[0061] Using the sucrose density gradient method (45%, 35%, 25%, 15%), the standard control antigen of type A foot-and-mouth disease was purified, and the content of 146S antigen was determined to be 1.74 μg / ml;

[0062] 3) To establish an indirect sandwich ELISA method for type A foot-and-mouth disease:

[0063] details as follows:

[0064] a) Coating ELISA plate: dilute FMD A rabbit antiserum (provided by Lanz...

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Abstract

The invention discloses a quantitative ELISA (Enzyme Linked Immunosorbent Assay) detection method for an antigen 146S of foot and mouth disease A. The method comprises the steps of (1) storing a standard control antigen of the foot and mouth disease A; (2) purifying and quantifying a standard control antigen 146S of the foot and mouth disease A; (3) establishing an indirect sandwiched ELISA method for the foot and mouth disease A; (4) drawing a standard curve of the standard control antigen; (5) counting the content of the antigen 146S of a tested sample; and (6) detecting sensitivity and specificity. The invention further provides a kit utilizing the detection method. The detection method and the kit have the advantages that when the quantitative ELISA detection method for the antigen 146S of the foot and mouth disease A is used for detecting the content of the foot and mouth disease antigen, not only can the content of the antigens be calculated, but also the antigens can be classified; and the kit prepared by utilizing a principle of the detection method has the advantages of sensitivity, specificity, simplicity in operation, time and labor conservation and good stability and is suitable for batch detection, and serotypes can be distinguished.

Description

technical field [0001] The invention relates to a type A foot-and-mouth disease 146S antigen quantitative ELISA detection method, a kit and application thereof. Background technique [0002] Foot and mouth disease (FMD) is an acute, febrile and highly contagious infectious disease common to major domestic animals such as pigs, cattle, sheep, and other cloven-hoofed animals, with more than 70 susceptible animals. The clinical feature is a vesicular eruption on the oral mucosa, hooves, and breast skin. The disease has many transmission routes and fast speed, and has repeatedly broken out in the world, causing huge political and economic losses. The World Organization for Animal Health (OIE) lists it as an infectious disease that must be declared, and my country also lists it as the first class of animal infectious diseases. [0003] Foot and mouth disease virus (FMDV) belongs to the genus FMDV of the picornaviridae family, and has seven serotypes: O, A, Asian I, C, and South...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
Inventor 马军武冯霞周广青杨亚民
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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