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Brucella latex microsphere antibody detection card and preparation method thereof

A technology of latex microspheres and Brucella, applied in the field of Brucella latex microsphere antibody detection card and its preparation, can solve the problems of narrow application range of Brucella antibody detection card, unstable product quality, high cost of raw materials, etc. Achieve the effects of low preparation cost, improved application range and high sensitivity

Pending Publication Date: 2022-01-11
SHANDONG VOCATIONAL ANIMAL SCI & VETERINARY COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The main technical problem to be solved by the present invention is to provide a Brucella latex microsphere antibody detection card and a preparation method thereof, which can solve the problem that the cost of raw materials for preparing the Brucella antibody detection card is too high at present; The problem of narrow application range of antibody detection card; solve the problem of uneven particle size in the firing of colloidal gold in the preparation process of traditional colloidal gold detection card, resulting in unstable product quality and poor uniformity

Method used

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  • Brucella latex microsphere antibody detection card and preparation method thereof
  • Brucella latex microsphere antibody detection card and preparation method thereof
  • Brucella latex microsphere antibody detection card and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0062] Example: see Figure 1-2 , a Brucella latex microsphere antibody detection card, comprising a shell 8 and a supporting sample diluent, the shell 8 is equipped with a test strip.

[0063] The test strip includes a PVC bottom plate 2, on which a sample pad 1, a marking pad 3, a coating film 6 and a water-absorbing pad 7 are pasted in sequence from one side of the PVC bottom plate 2 to the other.

[0064] The coating membrane 6 is a nitrocellulose membrane.

[0065] In this embodiment, the coating membrane 6 is a nitrocellulose membrane of model CN140 produced by Sartorius Company.

[0066] The coating film 6 is coated with a detection line 4 and a quality control line 5, the detection line 4 is coated at a position close to the marking pad 3, and the quality control line 5 is coated at a position close to the water-absorbing pad 7 .

[0067] The detection line 4 is coated with Brucella lipopolysaccharide (LPS), and the quality control line 5 is coated with a mouse anti...

Embodiment 2

[0120] Example 2: In the above-mentioned Example 1, the determination of the optimal pH value for protein labeling: During the protein labeling process of latex microspheres, MES solutions with pH values ​​of 8.0, 8.5 and 9.0 were used for labeling respectively, and different pH values ​​were compared. The influence of the specificity and sensitivity of the Brucella latex microsphere antibody detection card, so as to determine the optimal pH value of protein labeling, the experimental structure is as follows and Figure 8 shown.

[0121] The following table shows the experimental results of different marked pH values:

[0122]

[0123] In the above table, "+" means positive; "-" means negative.

[0124] According to the above table and Figure 8 It can be seen that when the pH value is 8.5, the detection line 4 of the Brucella latex microsphere antibody detection card in the present invention has better sensitivity, and the detection line 4 has better color development, a...

Embodiment 3

[0125] Embodiment 3, in above-mentioned embodiment 1, the selection of optimal protein labeling amount: in the protein labeling process of latex microsphere, adopt respectively marker protein and latex microsphere mass ratio to be 1:15,1:10,1: The ratio of 5 is marked, and the experimental structure is shown in the following table and Figure 9 shown.

[0126] The following table shows the experimental results of different protein labeling amounts:

[0127]

[0128] In the above table, "+" means positive; "-" means negative.

[0129] According to the above table and Figure 9 It can be seen that when the mass ratio of labeled protein to latex microspheres is 1:15, the sensitivity is poor, and the color of the detection line is weak; when the mass ratio is 1:10, the sensitivity and specificity are better, and the mass ratio is At 1:5, the color rendering effect of the Brucella latex microsphere antibody detection card in the present invention has no obvious change compare...

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Abstract

The invention belongs to the technical field of biological detection, and discloses a brucella latex microsphere antibody detection card and a preparation method thereof. The brucella latex microsphere antibody detection card comprises a shell and a sample diluent used in cooperation with the shell, wherein a test strip is assembled in the shell and comprises a PVC bottom plate, a sample pad, a marking pad, a coating film and a water absorption pad are adhered to the PVC bottom plate, and the marking pad is a glass cellulose film and is coated with a coupling marker of recombinant streptococcus protein G and latex microspheres; and a coating membrane is a nitrocellulose membrane, and the coating membrane is coated with a detection line coated with brucella lipopolysaccharide and a quality control line coated with a mouse anti-His tag monoclonal antibody. The brucella latex microsphere antibody detection card is low in preparation cost, is convenient and rapid, has the advantages of no need of equipment and instruments and no need of professionals, can be simultaneously suitable for detecting brucella antibodies in serum or blood samples of cattle, sheep and pigs, and widens the application range.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a Brucella latex microsphere antibody detection card and a preparation method thereof. Background technique [0002] Brucella (Brucella) is an intracellular parasitic small ball-shaped bacterium, Gram-staining negative, non-capsulated, positive for catalase and oxidase, and can survive in many kinds of livestock; Brucellosis (hereinafter referred to as Brucellosis) is a zoonotic infectious disease caused by bacteria of the genus Brucella. Animal brucellosis is mostly sheep and cattle. Its main clinical features are abortion, mastitis, infertility of livestock and various tissues (such as testis, joints) inflammation. [0003] Brucellosis is widely prevalent at home and abroad. So far, more than 170 countries and regions have reported brucellosis, which seriously threatens human health and affects the development of animal husbandry. It is an infectious d...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/58G01N33/577G01N33/558G01N33/543G01N33/532
CPCG01N33/56911G01N33/558G01N33/54313G01N33/577G01N33/585G01N33/532G01N2333/23G01N2469/20
Inventor 肖发沂王小伟齐艳君亓鹏朱光陈焱柏程昊
Owner SHANDONG VOCATIONAL ANIMAL SCI & VETERINARY COLLEGE
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