Bovine Brucella colloidal gold antibody detection test paper strip

A technology for detection of Brucella bovis and antibodies, which is applied in measurement devices, instruments, scientific instruments, etc., can solve problems such as non-specificity and lack of clear description of core technical characteristics, and achieves improved specificity, reduced non-specific reactions, and improved The effect of purity

Inactive Publication Date: 2015-12-09
CHINA INST OF VETERINARY DRUG CONTROL
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For another example, Hangzhou Dien Technology Co., Ltd. Zhang Mingzhou et al. (application number: CN201310033074.0) adopt Brucella bacterium antigen as quality control line coating antigen, and then utilize a kind of monoclonal antibody against erythrocyte as competition usefulness Monoclonal antibody, adsorbed and dried colloidal gold protein (flow band), established a sandwich test card for detecting Brucella bovis antigen. On the one hand, the patent did not clearly describe its core technical features. On the other hand, due to Rutella has strong bacterial antigen similarities with some Gram-negative bacteria, especially Yersinia O9 and Escherichia coli O157, and the patent uses Brucella bacterial antigens as colloidal gold particles Adsorbed antigens, in theory, the detection results will have certain non-specificity

Method used

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  • Bovine Brucella colloidal gold antibody detection test paper strip
  • Bovine Brucella colloidal gold antibody detection test paper strip
  • Bovine Brucella colloidal gold antibody detection test paper strip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] ——Study on the effect of different conditions on the size and color of colloidal gold particles

[0102] in 99mlH 2 O, add 1ml of 1% auric acid chloride (HAuCl4), so that the final concentration of auric acid chloride 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.

[0103] 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...

Embodiment 2

[0107] ——Sensitivity study of colloidal gold test strips

[0108] 1. Sensitivity test to the brucella bovis antibody positive control serum of gradient dilution After the brucella bovine antibody positive control serum (titer is 320IU) is carried out 2 times of gradient dilutions, test with 3 batches of lab-made colloidal gold The paper strip was tested to determine the sensitivity of the test strip to the gradiently diluted positive control serum, and the judgment time was within 4 minutes. The test results are shown in 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.

[0109] Table 2 Sensitivity detection of serially diluted positive control serum

[0110]

[0111] 2. Sensitivity test to known positive samples Three batches of colloidal...

Embodiment 3

[0116] ——Research on the specificity of colloidal gold test strips

[0117] 1. Specific detection of Yersinia enterocolitica O9 positive serum

[0118] Three batches of Brucella bovis colloidal gold test strips prepared by our laboratory were used to detect the positive serum of Yersinia enterocolitica O9. As a result, no specific band appeared at the position of the detection line within 3 minutes. is negative.

[0119] 2. Specific detection of known negative sera

[0120] 132 samples of Brucella bovis antibody-negative serum with known background were tested with 3 batches of colloidal gold test strips. As a result, 5 of the 3 batches of kits were not detected, and the serum numbers were 12, 34, 66, and 91 respectively. and 108, the specificity is 95.5%, the test results are shown in Table 4.

[0121] Table 4 The specificity results of different batches of kits to negative bovine serum

[0122]

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Abstract

The present invention relates to a rapid, efficient and accurate bovine Brucella colloidal gold diagnostic method. According to the present invention, Brucella strain S2 lipopolysaccharide (LPS) is adopted as a testing line coating antigen so as to improve the detection sensitivity; the LPS purification and purity determination method is improved, such that the developed kit has good specificity; the monoclonal antibody on the Fc terminal of the mouse anti-bovine IgG is used to label the colloidal gold to prepare the colloidal gold pad of the colloidal gold antibody detection test paper strip so as to further improve the detection specificity; the bovine Brucella colloidal gold antibody detection test paper strip can be used for detecting the Brucella antibody in the bovine serum, and has characteristics of strong specificity, high sensitivity, convenience, rapidness, and the like; and the complex detection equipment is not required for the Brucella detection, and the bovine Brucella colloidal gold antibody detection test paper strip is especially suitable for cattle farm breeding staffs and grassroots veterinarians.

Description

[0001] Technical Field The present invention relates to a diagnostic method for Brucella bovis antibody—Brucella bovis colloidal gold antibody detection test strip, 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 th...

Claims

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Application Information

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IPC IPC(8): G01N33/569
CPCG01N33/56911G01N2333/23
Inventor 丁家波蒋卉徐磊张阁张玉洁朱良全王楠毛开荣
Owner CHINA INST OF VETERINARY DRUG CONTROL
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