Quantitative enzyme-linked immuno-sorbent assay (ELISA) detection method for Asia I-type foot-and-mouth disease antigens 146S as well as kit and application of quantitative ELISA detection method

A detection kit and detection method technology, applied in measurement devices, color/spectral property measurement, instruments, etc., can solve problems such as being unsuitable for large-scale sample detection, complicated operation, expensive instruments, etc., to optimize vaccine production process, stability Good, easy-to-use effects

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
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Method used

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  • Quantitative enzyme-linked immuno-sorbent assay (ELISA) detection method for Asia I-type foot-and-mouth disease antigens 146S as well as kit and application of quantitative ELISA detection method
  • Quantitative enzyme-linked immuno-sorbent assay (ELISA) detection method for Asia I-type foot-and-mouth disease antigens 146S as well as kit and application of quantitative ELISA detection method
  • Quantitative enzyme-linked immuno-sorbent assay (ELISA) detection method for Asia I-type foot-and-mouth disease antigens 146S as well as kit and application of quantitative ELISA detection method

Examples

Experimental program
Comparison scheme
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Example Embodiment

[0056] Example 1:

[0057] A quantitative ELISA method for detecting Asian foot-and-mouth disease 146S antigen includes the following steps:

[0058] 1) Reserve the Asian Type I Foot-and-Mouth Disease Standard Control Antigen:

[0059] For Asian type I foot-and-mouth disease standard control antigen, one part is stored at -70℃ (thawed only once, for ELISA test) at 5 mL / bottle; the other part is stored at -70℃ at 3×100 mL (thawed only once, Used for 146S antigen purification and quantification (three times);

[0060] 2) Purification and quantitative Asian foot-and-mouth disease type I standard control antigen 146S:

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

[0062] 3) Establish an indirect sandwich ELISA method for Asian foot-and-mouth disease type I:

[0063] details as follows:

[0064] a) Coated ELISA plate: Dilute th...

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Abstract

The invention discloses a quantitative enzyme-linked immuno-sorbent assay (ELISA) detection method for Asia I-type foot-and-mouth disease antigens 146S. The method comprises the following steps of: (1) storing Asia I-type foot-and-mouth disease standard reference antigens; (2) purifying and quantifying the Asia I-type foot-and-mouth disease standard reference antigens 146S; (3) establishing an Asia I-type foot-and-mouth disease indirect sandwich ELISA method; (4) drawing a standard curve of the standard reference antigens; (5) calculating the content of the antigens 146S in a detected sample; and (6) detecting the sensitivity and specificity. The invention also provides a kit which adopts the detection method. The quantitative ELISA detection method for the Asia I-type foot-and-mouth disease antigens 146S and the kit have the beneficial effects that when the content of the foot-and-mouth disease antigens is measured by using the quantitative ELISA detection method, the content of the antigens can be calculated, and the antigens can be parted; and the kit prepared by using the principle of the quantitative ELISA detection method is sensitive, specific, simple to operate, good in stability and suitable for batch detection, saves time and labor, and can distinguish serotypes.

Description

technical field [0001] The invention relates to an Asian type I 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, an...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N33/569G01N33/544G01N33/535
Inventor 冯霞马军武周广青杨亚民
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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