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A composition and its application in the determination of chicken infectious bronchitis antibody

A bronchitis and antibody technology, applied in biological tests, immunoassays, measurement devices, etc., can solve the problems of cumbersome IBHI antibody detection process, shorten the preparation time, stabilize the measurement results, and simplify the sample measurement steps.

Active Publication Date: 2018-10-16
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problem that the serum to be tested must be treated with 25% kaolin suspension in the prior art to remove the non-specific hemagglutination inhibitory factor in the serum to be tested, and to improve the cumbersome detection process of IB HI antibody

Method used

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  • A composition and its application in the determination of chicken infectious bronchitis antibody
  • A composition and its application in the determination of chicken infectious bronchitis antibody
  • A composition and its application in the determination of chicken infectious bronchitis antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1 screens suitable diluent

[0026] The serum to be tested was not treated with 25% kaolin suspension, and four different dilutions of normal saline, HA buffer, 0.01mol / L PBS pH 7.2 and 1 / 15mol / L PBS pH 7.2 were selected to determine the effect on the level of IB HI antibody (The whole process of the test, including the preparation of 1% chicken erythrocyte suspension, was carried out with the same corresponding diluent). Using the 20140625 batch of IBHI antigen (prepared by our laboratory and obtained the national second-class new veterinary drug registration certificate), using physiological saline, HA buffer, 0.01mol / L PBS pH 7.2 and 1 / 15mol / L PBS pH 7.2 The antibody titers of 8 different chicken sera were determined. The test results showed that 0.01mol / L pH 7.2PBS and 1 / 15mol / L pH 7.2PBS were used as diluents for determination, the two IB positive sera were both positive, and the HI titer was 10-11log2; SPF chicken serum, IB Negative serum, EDS positiv...

Embodiment 2

[0027] Embodiment 2 optimizes the dilution obtained by screening

[0028] The serum to be tested does not need to be treated with 25% kaolin suspension, but 0.01mol / L PBS dilutions with three different pH values ​​of pH 6.0, pH 6.6 and pH 7.2 are selected to determine the impact on the IB HI antibody level (the whole process of the test includes 1% chicken The preparation of the red blood cell suspension was carried out with the corresponding diluent). Using 20140625 batch of IB HI antigen, using three different 0.01mol / L PBS dilutions of pH 6.0, pH 6.6 and pH 7.2, the antibody titers of 8 different chicken sera were determined. The test results showed that the 2 IB positive sera were positive when measured with 0.01mol / L PBS pH 7.2, the HI titer was 9-11log2, and the SPF chicken serum and H9 subtype avian influenza (Q strain) were positive Serum, H9 subtype avian influenza (WD strain) positive serum, EDS positive serum and ND positive serum were all negative, and the HI tite...

Embodiment 3

[0029] Example 3 Diluent Sensitivity Verification

[0030]Using three batches of IB HI test antigens prepared by 20130116, 20140625 and 20140722 laboratories, using a new method of 0.01mol / L PBS buffer with pH 6.6 as the diluent, 86 chicken serum samples were tested for IB HI antibody levels , to observe the sensitivity of the new method in detection. 86 chicken serum samples including 1 IB positive control serum, ND positive serum, H9 subtype avian influenza positive serum, H5 subtype avian influenza positive serum, EDS positive serum and SPF chicken serum. 40 copies of chicken sera for partial effect test; 40 copies of IBV M41 strain infected chicken serum (sera 21 days after IBV M41 strain infected SPF chicken). The results showed that among the 86 samples tested, 80 were known to be immune or infected chicken sera, and 66 positive samples were detected, with a positive detection rate of 82.5% (66 / 80). Among them, 20 copies of chicken first-immune serum (21 days after H12...

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Abstract

The invention provides a specific composition, which is prepared from sodium dihydrogen phosphate, disodium hydrogen phosphate and sodium chloride, wherein the mass ratio of the sodium dihydrogen phosphate to the disodium hydrogen phosphate to the sodium chloride is 1.065:1.5:(11.7 to 34). The composition is adopted to prepare a diluent for determining an infectious bronchitis antibody. According to the composition provided by the invention, the necessary step of processing detected sera by 25 percent of kaolin suspension to remove non-specificity hemagglutination inhibition factors of the detected sera during the infectious bronchitis antibody determination process in the prior art is skipped, and an infectious bronchitis antibody detection technology is improved. The diluent provided by the invention is adopted to determine the infectious bronchitis antibody, so that the previous test preparation time can be shortened, the sample determination step is simplified, the determination result is stable, and the diluent is suitable for infectious bronchitis antibody determination.

Description

technical field [0001] The invention relates to the field of immune analysis and determination, in particular to the application of a composition to the determination of chicken infectious bronchitis antibodies. Background technique [0002] At present, the existing chicken infectious bronchitis antibody level determination technology is according to the People's Republic of China veterinary biological products quality standard "chicken infectious bronchitis virus (M41 strain) hemagglutination inhibition test antigen, positive serum and negative serum" It is stipulated that 25% kaolin suspension is used to treat the serum to be tested, and the diluent is HA buffer solution. The test results show that the HI antibodies of infectious bronchitis (IB) positive serum are all positive, and SPF chicken serum, IB negative serum and other chicken serum are all positive. The IB HI titers of the positive sera were all ≤3log2, which met the requirements of the regulations. [0003] How...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/569G01N33/531
CPCG01N33/531G01N33/56983G01N33/6854G01N2469/20G01N2800/127
Inventor 姜北宇章振华李林沈佳张建伟史爱华景小冬郑小兰黄凤军
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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