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Method for enhancing detection reagent sensitivity by using colloidal gold or latex granule as marker

A technology of latex particles and colloidal metals, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of pathogens affecting the application of reagents and low detection sensitivity, and achieve the effect of improving specificity and increasing sensitivity

Inactive Publication Date: 2008-08-06
储宁
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

, it is precisely because the detection is fast and the reaction time of antigen and antibody is short, resulting in low detection sensitivity
[0034] Especially when used in the field of clinical testing, due to the low content of antigens (such as viruses, bacteria, etc.) in the lesions of patients, many test papers or percolation kits made with colloidal metal or latex particles as markers cannot accurately detect pathogens The presence of these reagents affects the application of

Method used

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  • Method for enhancing detection reagent sensitivity by using colloidal gold or latex granule as marker
  • Method for enhancing detection reagent sensitivity by using colloidal gold or latex granule as marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: Respiratory syncytial virus latex method detection kit

[0054] (1) Purchase latex particles with suitable particle size.

[0055] (2) Marking of latex particles

[0056] Label according to the steps in the instruction manual of the latex particles, and label the anti-respiratory syncytial virus monoclonal antibody on the latex particles. Centrifuge and purify the labeled latex particles at an appropriate speed, carefully suck off the supernatant, and suspend the precipitate with preservation solution to make latex particle-antibody conjugates.

[0057](3) Drying of latex particle-antibody conjugate

[0058] Take the latex particle-antibody conjugate solution, dilute it to an appropriate concentration with the working solution, and distribute it into 2 ml vials or small tubes. Freeze-dry overnight in a freeze dryer until completely dry. After sealing, keep airtight and dry.

[0059] (4) Antibody kit for nitrocellulose membrane

[0060] ① Dilute the anti...

Embodiment 2

[0083] Embodiment 2: A and B influenza virus colloidal gold detection kits

[0084] (1) Preparation of colloidal gold Colloidal gold particles of 20 to 50 nm were prepared by chemical reduction hair or other suitable methods.

[0085] (2) Marking of colloidal gold

[0086] The colloidal gold solution prepared in step (1) was treated with 0.2M K 2 CO 3 Adjust to a suitable pH value. Add the anti-influenza A virus monoclonal antibody into the above solution in an appropriate labeled amount, and stir at room temperature. Add stabilizers such as BSA and PEG of appropriate concentration, and stir at room temperature. Centrifuge at an appropriate speed until the supernatant is basically colorless, carefully suck off the supernatant, and suspend the precipitate with colloidal gold preservation solution.

[0087] The colloidal gold solution prepared in step (1) was treated with 0.2M K 2 CO 3 Adjust to a suitable pH value. Add the anti-influenza B virus monoclonal antibody into...

Embodiment 3

[0117] Embodiment 3: Treponema pallidum colloidal selenium detection kit

[0118] (1) Purchase or prepare colloidal selenium solution with suitable particle size.

[0119] (2) Labeling of colloidal selenium

[0120] Follow the appropriate steps for labeling and label the anti-Treponema pallidum monoclonal antibody onto the colloidal selenium particles. Centrifuge and purify the labeled latex particles at an appropriate speed, carefully suck off the supernatant, and suspend the precipitate with the preservation solution to prepare the colloidal selenium-antibody conjugate.

[0121] (3) drying of colloidal selenium-antibody conjugate

[0122] Take the colloidal selenium-antibody conjugate solution and dilute it with working solution to an appropriate concentration.

[0123] Spread the glass fiber or PE composite membrane on a glass plate, and evenly drop the diluted colloidal selenium-antibody conjugate solution on it. Freeze-dry overnight in a freeze dryer until completely ...

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Abstract

The invention discloses a method capable of improving the sensitivity of a quick detection reagent which is prepared with labels made of colloidal metal (colloidal gold, colloidal selenium, etc.) or emulsion grains. The application range of the method is the immune chromatographic detection test paper or the percolation reagent which is prepared with labels made of colloidal metal (colloidal gold, colloidal selenium, etc.) or emulsion grains and adopts a double antibody sandwich principle. The method is characterized in that: the colloidal metal or emulsion grains which is labeled with the antibody and undergoes drying is not adhered between a sample cushion and a cellulose nitrate film and is dried on other containers or supporters; during detection, firstly the colloidal metal or emulsion grains which is labeled with the antibody and undergoes drying is mixed with the sample to be detected. Only the sample cushion, the cellulose nitrate film, and a water absorption cushion are adhered on the test paper scrip, wherein the sample cushion is used to contain the sample to be detected, the cellulose nitrate film is provided with fixed antibody to serve as detection line, the number of the detection line can be one or more than one, and a comparison line is also fixed on the cellulose nitrate film.

Description

technical field [0001] The invention relates to a method capable of improving the sensitivity of a rapid detection reagent made of colloidal metal (colloidal gold, colloidal selenium, etc.) or latex particles as a marker. (Colloidal gold, colloidal selenium, etc.) or latex particles as immunochromatographic test paper or diafiltration reagent made as markers. Background technique [0002] In 1971, Faulk and Taytor introduced colloidal gold into the field of immunochemistry. Since then, immunocolloidal gold technology has been widely used in various fields of biomedicine as a new immunological method. The current applications in medical testing are mainly immunochromatography (immunochromatography-phy) and rapid immunogold filtration assay (Dot-immuogold filtration assay DIGFA), which are used to detect pathogens such as HBsAg, HCG, bacteria, and viruses. Fast, accurate and non-polluting advantages. [0003] (1) Basic principle of immune colloidal gold technology [0004] ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/532
Inventor 储宁
Owner 储宁
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