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Peste des petits ruminants virus IgG antibody colloidal gold detection card and production and using methods thereof

A ruminant virus, colloidal gold technology, applied in the field of immunology, to achieve the effect of simple operation, stable results and low price

Inactive Publication Date: 2011-03-30
深圳市三方圆生物科技股份有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the monitoring of Peste des petits ruminants by this technology at home and abroad

Method used

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  • Peste des petits ruminants virus IgG antibody colloidal gold detection card and production and using methods thereof
  • Peste des petits ruminants virus IgG antibody colloidal gold detection card and production and using methods thereof
  • Peste des petits ruminants virus IgG antibody colloidal gold detection card and production and using methods thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The preparation of embodiment 1 colloidal gold particle

[0033] Get 1% chloroauric acid solution 1ml, add 99ml ultrapure water to form a chloroauric acid solution with a final concentration of 0.01%, after heating and boiling, get 1.6ml of 1% trisodium citrate and add it to the boiling chloroauric acid solution quickly at one time, Continue heating until the solution turns from light yellow to blue-black and finally bright red. After the color is stable, continue heating for 5 minutes, cool at room temperature, and replenish the lost water to the original volume.

Embodiment 2

[0034] Preparation of embodiment 2 colloidal gold-labeled staphylococcal protein A complex:

[0035] Labeling can be performed when the optimum stable amount of protein and the optimum pH value for labeling are determined. The specific steps are as follows: calculate the total amount of protein to be labeled according to 120% of the minimum stable amount. Under magnetic stirring, add the protein solution into the colloidal gold solution (pH has been adjusted to 5.9-6.2). When adding the protein, it should be added drop by drop, and 1 mg of protein should be added in about 5 minutes. Take 1ml of colloidal gold-staphylococcus protein A conjugate solution (experimental group) and 1ml of colloidal gold stock solution (control group) respectively, add 0.1ml of 10% sodium chloride solution to the test tube, and let stand at room temperature for 1h, and observe the results: if the control group The test tube solution turned from red to blue, and even polymer precipitation can be see...

Embodiment 3

[0036] Example 3 Purification of colloidal gold-labeled staphylococcus protein A complex:

[0037] Purification of immunocolloidal gold complexes by ultracentrifugation. First centrifuge at 3500rpm for 20min (4°C), discard the precipitate, centrifuge the supernatant at 13500rpm for 35min (4°C), discard the supernatant, suspend the loose red sediment with preservation solution; then centrifuge at 11000rpm for 35min (4°C), After carefully discarding the supernatant, suspend the loose red sediment to 1 / 10 of the original volume with the preservation solution, which is the initially purified immunocolloidal gold complex.

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Abstract

The invention relates to a peste des petits ruminants virus IgG antibody colloidal gold detection card and production and using methods thereof, and belongs to the technical field of immunology. The rapid diagnostic card comprises a shell and test paper, and is characterized in that: the shell is provided with a sample feeding hole and an observation window; the test paper is arranged in the shell; nitrocellulose films are stacked in the middle of a PVC lining plate, and water absorption pads and sample pads are respectively stacked at two ends; the nitrocellulose films are lapped with the water absorption pads and the sample pads; a section of membrane containing a staphylococcal protein A immune colloidal gold compound is additionally arranged in a sample area; and a detection belt containing inactivated peste des petits ruminants virus antigen and a quality control belt containing goat anti-rabbit IgG are formed on the nitrocellulose films. The invention provides a rapid, accurate and sensitive diagnostic card without special instruments, which is easy to operate, and can meet the detection requirements of food safety, slaughter and detection mechanisms.

Description

technical field [0001] The invention relates to a small ruminant virus IgG antibody colloidal gold detection card, which belongs to the technical field of immunology. Background technique [0002] Peste des petits ruminants is caused by Peste des petits ruminants virus (PPRV) of the family Paramyxoviridae. It is an acute contagious disease characterized by fever, stomatitis, diarrhea and pneumonia. Goats and sheep are susceptible, and the incidence of goats and The case fatality rate is high. The World Organization for Animal Health (OIE) lists it as a legally notifiable animal disease, and my country lists it as a first-class animal disease. The disease first occurred in Côte d'Ivoire in Africa in 1942. In 2003, 25 countries reported the occurrence of Peste des petits ruminants, mainly distributed in Asia, Africa, and Turkey in Europe. Since the outbreak of Peste des petits ruminants in Ritu County, Tibet Autonomous Region on July 25, 2007 in my country, many epidemics ha...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/558G01N33/532
Inventor 钟松清刘忠清谭攀
Owner 深圳市三方圆生物科技股份有限公司
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