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Colloidal gold test paper bar for antibody detection of sheep Brucella

A technology for detecting Brucella sheep and antibodies, which is applied to measuring devices, instruments, scientific instruments, etc., to achieve the effects of eliminating interference, improving purity, and reducing non-specific reactions

Inactive Publication Date: 2015-12-16
CHINA INST OF VETERINARY DRUG CONTROL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no colloidal gold test strip specifically for the detection of brucellosis

Method used

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  • Colloidal gold test paper bar for antibody detection of sheep Brucella
  • Colloidal gold test paper bar for antibody detection of sheep Brucella
  • Colloidal gold test paper bar for antibody detection of sheep Brucella

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] ——The effect of different conditions on the size and color of colloidal gold particles

[0104] in 99mlH 2 O, add 1ml 1% chloroauric acid (HAuCl 4 ), so that the final concentration of chloroauric acid is 0.01%. Continue heating the chloroauric acid aqueous solution to boiling, add 1.05% trisodium citrate aqueous solution (reducing agent) with different volumes of 1.0ml, 1.2ml, 1.4ml, 1.6ml, 1.8ml, 2.0ml, continue heating for 5min, and use Cool in an ice water bath. The prepared colloidal gold solution was stored at 4°C in the dark. Take a small amount of cooled colloidal gold solution and measure the dispersion and homogeneity of the colloidal gold solution with an ultraviolet-visible spectrometer, and observe the particle size of the gold particles with a transmission electron microscope.

[0105] The result was that the amount of the original agent added was inversely proportional to the particle size of the colloidal gold, that is, the greater the input amount o...

Embodiment 2

[0109] ——Sensitivity of colloidal gold test strips

[0110] 1. Sensitivity test to serially diluted Brucella sheep antibody positive control serum

[0111] After the Brucella sheep antibody positive control serum (titer is 320IU) is carried out 2 times of gradient dilution, detect with 3 batches of laboratory self-made colloidal gold test strips, determine that this test strip is to the positive control serum of gradient dilution. Sensitivity, the judgment time is within 4 minutes, see the test results Table 2 . The results show that the colloidal gold test strip involved in the present invention has good sensitivity, and can detect the Brucella positive serum with a titer of 2.5IU, which is higher than that of the tiger bengal plate agglutination test and the complement fixation test.

[0112] Table 2 Sensitivity assay against serially diluted positive control sera

[0113]

[0114] 2. Sensitivity test on known positive samples

[0115] Three batches of colloidal go...

Embodiment 3

[0119] ——Study on the specificity of colloidal gold test strips

[0120] 1. Specific detection of Yersinia enterocolitica O9 positive serum

[0121] Three batches of Brucella sheep colloidal gold test strips prepared by our laboratory were used to detect Yersinia enterocolitica O9 positive serum. As a result, no specific band appeared at the position of the detection line within 4 minutes. Negative.

[0122] 2. Specific detection of known negative sera

[0123] 71 samples of Brucella sheep antibody-negative serum with known background were tested with 3 batches of colloidal gold test strips. As a result, 3 samples of the 3 batches of kits were not detected, and the serum numbers were 7, 91 and 58, respectively. The test results are 95.8%, see Table 4 .

[0124] Table 4 Specific results of different batches of kits for negative bovine serum

[0125]

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Abstract

The invention relates to a colloidal gold test paper bar for antibody detection of sheep Brucella. In the invention, lipopolysaccharide (LPS) extracted from a Brucella vaccine strain (S2) is taken as an envelope antigen of the colloidal gold test paper bar and improves the sensitivity of detection. Monoclonal-antibody-labeled colloidal gold of a mouse anti-sheep IgGFc terminal is prepared into a gold colloid pad of the colloidal gold test paper bar for antibody detection, and nonspecific adsorption of antibody protein by SPA in the tradition is changed. The monoclonal-antibody-labeled colloidal gold is taken as a colloidal gold labeled protein, so that specificity of detection is further improved. A set of serum for controlling quality of the test paper bar is prepared, and specificity and sensitivity of the test paper bar is ensured. The test paper bar can be used for on-site sampling, is convenient and rapid to use, and is quite suitable for application to a basic farm or a basic veterinarian.

Description

[0001] Technical field The present invention relates to a test strip for colloidal gold antibody detection of Brucella sheep, which belongs to the technical field of biological product detection. technical background [0002] Brucellosis (brucellosis) is a zoonotic disease characterized by abortion and fever caused by Brucella or Brucella, which seriously threatens the life and health of humans and various animals. . This disease not only has serious harm to the reproduction and production performance of animals, but more importantly, people are often difficult to cure after being infected with Brucella, thus causing serious public health problems. Elimination of brucellosis has therefore been one of the most important goals of public health programs in countries where Brucella is endemic. [0003] The existence of brucellosis in the world has a long history. Human beings have gradually deepened their understanding of the disease, and their diagnosis level has been continuous...

Claims

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Application Information

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IPC IPC(8): G01N33/569
CPCG01N33/56911G01N2333/23
Inventor 丁家波张玉洁蒋卉徐磊张阁王楠朱良全毛开荣
Owner CHINA INST OF VETERINARY DRUG CONTROL
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