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Sample treatment agent used on solid phase membrane immune analysis method mobile phase

An immunoassay and mobile phase technology, which is applied in the preparation of test samples, analysis materials, and material inspection products, etc., can solve the problems of reducing the detection sensitivity of samples to be tested, limiting the application of pathogenic microorganisms, and affecting the flow of test objects, etc., to achieve raw materials The effect of easy source, improved detection sensitivity, and simple preparation method

Inactive Publication Date: 2007-05-09
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the size of the particulate antigen will not only affect the flow of the detection object on the membrane, but also its spatial structure will inevitably limit its full combination with the capture antibody, thus reducing the detection sensitivity of the sample to be tested
In order to improve the detection sensitivity, it is necessary to dissociate and dissolve it from the particles to the greatest extent through effective chemical preparations without affecting its subsequent binding with the corresponding antibody. cannot meet this requirement
[0005] In addition, if the detection object is pathogenic microorganisms in complex samples, such as food, tissue samples, soil or white powder, etc., how to separate and detect the granular antigen to be tested from complex samples without or minimizing the influence of sample substances It is a very difficult problem. The current sample preparations used as carrier flow phases in immunochromatography systems cannot meet the above requirements. This is also the difficulty of the current solid-phase membrane immunoassay method applied to the detection of pathogenic microorganism particulate antigens such as bacteria. Serious Limits its application in the direct detection of pathogenic microorganisms

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Use PBS buffer as the solution matrix of the present invention, adjust its pH value to 8.0, and prepare a solution containing the following components:

[0018] Nonidet P40 3.5%(v / v)

[0019] The above sample preparation preparation is used to treat milk powder added with Yersinia pestis (approximately 50mg milk powder is added to each milliliter of sample preparation). After mixing for 5-10 minutes, centrifuge at 8000 rpm for 15 seconds. Colloidal gold immunochromatographic method and UCP (up-converting phosphorescence) immunochromatographic method were detected separately. The results show that compared with treatment with different amounts of general-purpose Tween20 solution, the detection sensitivity of the colloidal gold test can be increased by 5-10 times after the sample treatment preparation of this example is used. The sensitivity can be increased by more than 10 times. Similar comparison results can be obtained for the established UCP immunochromatographic method ...

Embodiment 2

[0021] Take PBS buffered salt solution as the solution matrix of the present invention, adjust its pH value to 7.5, and prepare a solution containing the following components:

[0022] Nonidet P40 1.0%(v / v)

[0023] NaN 3 (Sodium azide)0.05%(w / v)

[0024] Treat the above sample preparation preparations to Yersinia pestis, and use the same concentration of Tween20 (v / v), SDS (w / v), deoxycholate (w / v), Triton X-100 ( v / v) solution for comparison. After mixing for 5-10 minutes, use UCP (up-converting phosphorescence) immunochromatographic technique to detect Yersinia pestis. The comparison results show that for the up-converted phosphorescent labeling immunoassay, the sample preparation of this example has a higher positive detection value, and the detection sensitivity can be increased by at least 10 times. Furthermore, 0.05% NaN by weight is added to the solution matrix 3 , Can also play a role in corrosion protection, so that the prepared solution will not deteriorate.

Embodiment 3

[0026] Use PBS buffered salt solution as the solution matrix of the present invention, adjust its pH value to 5.5, and prepare a solution containing the following components:

[0027] Nonidet P40 0.5%(v / v)

[0028] NaN 3 (Sodium azide)0.05%(w / v)

[0029] BSA 1.0%(w / v)

[0030] The above-mentioned sample processing preparations can be used in existing immunochromatographic methods or immunofiltration tests, and testing and processing such as soil, milk powder, meat and bone meal, animal tissue, broth, and blood waiting for testing samples. After adding the sample treatment preparation and mixing for 5-10 minutes, leave it to stand or briefly centrifuge or coarse filter, insert the existing solid-phase membrane immunoassay test strip, or take the supernatant or filtrate and add an appropriate amount, and observe Test results. The test objects can be differentiated according to different test purposes, which can be bacteria, viruses, parasites or other meaningful detection ligand com...

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Abstract

The present invention disclosed a sample treatment preparation used for moving phase of solid-phase membrane immunoanalysis method. Said preparation includes basic buffer solution and Nonidet p40, in which the pH value of said basic buffer solution is 5.5-10, and the volume ratio content of Nonidet P40 is 0.05%-3.5%. Said invented preparation can effectively dissociate the particulate antigen components in complex samples such as milk powder, animal tissue and soil, etc.

Description

Technical field [0001] The invention relates to a sample processing preparation, in particular to a sample processing preparation used in the mobile phase of a solid-phase membrane immunoassay method. Background technique [0002] With the widespread implementation of the global biosafety strategy, the demand for rapid diagnosis of pathogenic organisms is increasing. At the same time, the detection targets for rapid diagnosis are more complex, including human, animal and plant samples, food, and even "white powder". Membrane-based immunoassay (Membrane-based immunoassay), especially immunochromatography technology, due to its simplicity, rapidity, high efficiency, low cost, and suitability for large-scale field applications, has made considerable progress in pathogen detection in recent years , Various new biomarker materials and optical and electronic detection systems have also made continuous progress and been rapidly applied. However, the technology for processing samples of ...

Claims

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Application Information

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IPC IPC(8): G01N1/28G01N33/531G01N33/53
Inventor 胡孔新王宝麟王大宁王静姚李四李伟陈维娜
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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