Detection method and detection kit used for detecting immune antibody of micro-encapsulation escherichia coli oral vaccine

A technology of detection kits and detection methods, which can be applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of unclear antigen sources, high cost and high cost, and achieve the effect of rapid and timely on-site monitoring and good monitoring.

Inactive Publication Date: 2016-07-13
XINJIANG AGRI UNIV
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

The disadvantage is that the primary antibody in the direct ELISA experiment must be labeled with an enzyme, but not every enzyme is suitable for labeling, and the cost is relatively high; the probability of interactive reactions in the indirect ELISA experiment is high
None of the antibodies currently detectable by commercial ELISA assays are specific to those produced by microencapsulated inactivated oral vaccines after stimulating the body
[0005] In addition, conventional ELISA detection methods for monitoring antibodies have the disadvantages of unclear source of antigens and high costs for large-scale identification and detection of antibody levels, and because microencapsulated E. There are no reports on the method of level detection and related demonstration methods

Method used

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  • Detection method and detection kit used for detecting immune antibody of micro-encapsulation escherichia coli oral vaccine
  • Detection method and detection kit used for detecting immune antibody of micro-encapsulation escherichia coli oral vaccine
  • Detection method and detection kit used for detecting immune antibody of micro-encapsulation escherichia coli oral vaccine

Examples

Experimental program
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Embodiment 1

[0016] Embodiment 1, be used for the detection method of microencapsulated Escherichia coli oral vaccine immune antibody detection, this detection method comprises the following steps: (1) antigen coating: be 9.6 carbonate buffer solution 0.05mol / L with pH Dilute the antigen according to a certain ratio, add 100 μl to each well, and coat at 4°C for 24 hours; (2) Washing: after taking out, equilibrate at room temperature for 30 minutes, discard the liquid, and use 1 times phosphate containing 0.05% Tween-20 at pH 7.4 Wash with buffer PBST 3 times, each time for 1 min, blot dry on absorbent paper; (3) Block: Add 100 μl of 1% BSA blocking solution per well, and incubate at 37°C for 1 h; (4) Wash: Discard the liquid, wash 3 times, Blot dry with absorbent paper; (5) add primary antibody: add 100 μl of standard positive serum diluted with 0.1% BSA diluent per well, incubate at 37°C for 1 hour, the primary antibody is the antibody in the serum to be tested; (6) wash : wash 3 times, b...

Embodiment 2

[0053] Embodiment 2. A detection kit for the detection of microencapsulated Escherichia coli oral vaccine immune antibody prepared according to the above detection method, which is applied to animals after microencapsulated Escherichia coli oral vaccine immunization, and can be aimed at the body produced Specific immune antibodies for detection.

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Abstract

The invention belongs to the technical field of research and development of a detection method and a detection kit of effect of a biological vaccine, and particularly relates to a detection method and a detection kit used for detecting an immune antibody of a micro-encapsulation escherichia coli oral vaccine. The detection method comprises the following steps: (1) coating an antigen; (2) washing; (3) sealing; (4) washing; (5) adding a first antibody; (6) washing; (7) adding a secondary antibody; (8) washing; (9) developing; (10) terminating; and (11) detecting an OD value by an enzyme-linked immunosorbent assay instrument. With the adoption of the detection method provided by the invention, the immune effect of the escherichia coli oral vaccine can be monitored very well; and when the detection kit with the method is applied, in-situ monitoring can be rapidly carried out in time.

Description

technical field [0001] The invention belongs to the technical field of detection methods and detection kits for biological vaccine effects, and in particular relates to a detection method and detection kits for the detection of serological antibody titers of microencapsulated oral vaccine immune effects. Background technique [0002] Sheep colibacillosis is diarrhea in young animals caused by different serotypes of pathogenic Escherichia coli, especially newborn young animals are particularly susceptible, and can lead to growth retardation, low production performance, and even death in severe cases. serious impact on economic development. At present, the prevention and treatment of this disease is mainly based on drug therapy and vaccination. Vaccination stimulates the body to produce antibodies, which can provide economical and effective immune protection for vaccinated livestock in animal husbandry, and can prevent corresponding infectious diseases at the same time. Epide...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56916
Inventor 姚刚徐新龙夏瑞阳马雪连王金泉钟旗苏艳雷程红
Owner XINJIANG AGRI UNIV
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