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Cocktail antigen for detection of allergic reaction of bovine tuberculosis

An allergy and cocktail technology, applied in the field of animal disease detection, can solve the problems of cumbersome, difficult to popularize and apply, and achieve the effects of simple detection process, specific diagnosis, and exclusion of non-specific mycobacterial infection.

Active Publication Date: 2019-05-21
CHINA ANIMAL HEALTH & EPIDEMIOLOGY CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is almost difficult to popularize and apply because it is too cumbersome to compare allergies

Method used

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  • Cocktail antigen for detection of allergic reaction of bovine tuberculosis
  • Cocktail antigen for detection of allergic reaction of bovine tuberculosis
  • Cocktail antigen for detection of allergic reaction of bovine tuberculosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Screening of Cocktail Antigens for Allergy

[0032] 1. Screening of antigenic components

[0033] The mixture of ESAT6 and CFP10 is called EC as an antigen, and its concentration is marked according to the concentration of a single component. The four antigens EC, FixB, MPB83 and HSPX were diluted to a final concentration of 10 μg / mL, and three groups of animals including naturally infected cattle, Mycobacterium avium-infected cattle and healthy cattle were used for allergy skin test.

[0034] When the average skin thickness difference between Mycobacterium avium-infected animals and healthy cattle detected by the four antigens was almost the same, the average skin thickness difference of naturally infected animals detected by EC antigen was greater than that of MPB83, FixB, and HSPX antigens. That is, when EC is used as a differential diagnosis antigen, the difference in skin thickness between naturally infected animals and non-specifically infected animals...

Embodiment 2

[0039] Example 2: Preparation of the cocktail antigen of the present invention according to the determined antigen combination and concentration

[0040] 1. Preparation method

[0041] (1) Test materials

[0042] 1. Main instruments

[0043] Constant temperature oscillator: Haimen Qilinbeier Instrument Manufacturing Co., Ltd.

[0044] Decolorization shaker: QILINBEIER company

[0045] Steady current and voltage electrophoresis instrument: Beijing Junyi Instrument Factory Co., Ltd.

[0046] Ultrasonic cell pulverizer: Ningbo Xinzhi Technology Biological Co., Ltd.

[0047] Trans-SD universal semi-dry transfer electrophoresis tank: Beijing Kaiyuan Xinrui Instrument Co., Ltd.

[0048] Ice machine: Shanghai Anting Scientific Instrument Factory

[0049] UV-Vis spectrophotometer: SHIMADZU

[0050] Trace Nucleic Acid Protein Analyzer: Nanodrop

[0051] Freeze Dryer: Toshiba

[0052] 2. Recombinant plasmid

[0053] The recombinant pET-28a-HSPX, pET-28a-FixB, pET-28a-ESAT6 and...

Embodiment 3

[0096] Example 3: Application of Antigen Cocktail for Allergy

[0097] The cocktail antigen prepared by the present invention is used as the antigen for bovine tuberculosis allergic reaction, and the specific steps are as follows:

[0098] 1. Preparation of antigen:

[0099] The purified ESAT6, CFP10, FixB and HSPX proteins were respectively concentrated to a concentration of 0.7 mg / mL using polyethylene glycol 20000, mixed according to the ratio of 2:2:1.5:1.5 to prepare the cocktail antigen of the present invention, and packed into 1mL / bottle, store at 4°C after freeze-drying.

[0100] 2. Allergy test

[0101] Before use, take 5 mL of LPBS, add it to the lyophilized antigen cocktail, mix well and use, so that the final concentrations of ESAT6, CFP10, FixB and HSPX are 40 μg / mL, 40 μg / mL, 30 μg / mL, and 30 μg / mL, respectively.

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Abstract

The invention provides a cocktail antigen for detection of an allergic reaction of bovine tuberculosis. The cocktail antigen comprises an ESAT6 antigen, a CFP10 antigen, an FixB antigen and an HSPX antigen, wherein the mass ratio of the ESAT6 antigen to the CFP10 antigen to the FixB antigen to the HSPX antigen is 2:2:1:5:1.5. The cocktail antigen is used for preparing a detection antigen of the allergic reaction of the bovine tuberculosis. The cocktail antigen can replace bovine type PPD and poultry type PPD used for comparing the allergic reaction, a single intradermal allergic reaction is carried out by using the cocktail antigen to achieve the effect of comparing the allergic reaction, a series of operation such as two-point shearing, measurement of the skin thickness, injection, re-testing of the skin thickness after 72 hours and calculation of a skin thickness difference are simplified into single-point operation and do not need calculation, the whole detection process is simplerand more convenient; meanwhile, the infection of poultry type mycobacteria and other non-specific mycobacteria can be excluded, and the bovine tuberculosis is diagnosed more specially.

Description

technical field [0001] The invention belongs to the technical field of animal epidemic detection, and in particular relates to a cocktail antigen used for detection of bovine tuberculosis allergic reaction. Background technique [0002] Bovine tuberculosis is a chronic consumptive zoonotic disease. It is an animal disease that must be notified according to the International Organization for Animal Health (OIE). It is a second-class animal infectious disease in my country. pose a major threat. According to statistics, about 5% of human tuberculosis is caused by Mycobacterium bovis. Therefore, it is of great public health significance to grasp the current situation of bovine tuberculosis in my country and to eliminate infected cattle through quarantine. The quarantine of bovine tuberculosis can be classified into the following three categories: bacteriological detection, molecular biological detection and immunological detection. Bacteriological detection and molecular biolog...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/35G01N33/68
Inventor 张喜悦孙明军李阳孙翔翔尼博陈爽魏荣
Owner CHINA ANIMAL HEALTH & EPIDEMIOLOGY CENT
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