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Immunochromatography assay and device taking color-developing agent as sample carrier and capable of repeated sample adding

An immunochromatography and color-developing reagent technology, applied in the field of immunochromatography, can solve the problems of decreased effective utilization rate, long time required to flow through the reaction zone, slow color reaction, etc., and achieves improved sensitivity and shortened interpretation time. Effect

Active Publication Date: 2013-03-20
益思美诠生物科技(上海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The sample amount of the latter method is slightly less than that of the former method, but because the sample is directly added to the solid-phase membrane first, due to the adsorption of the solid-phase membrane, the effective use rate of the sample will definitely decrease, and the sensitivity to the reaction will also decrease. will have a certain impact
In addition, since the chromogenic agent is added after the sample, it takes a long time for it to flow through the reaction zone, so the color reaction occurs slowly

Method used

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  • Immunochromatography assay and device taking color-developing agent as sample carrier and capable of repeated sample adding
  • Immunochromatography assay and device taking color-developing agent as sample carrier and capable of repeated sample adding
  • Immunochromatography assay and device taking color-developing agent as sample carrier and capable of repeated sample adding

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Embodiment 1 former method and method of the present invention detect HCV (hepatitis C virus) antibody

[0035] In this example, the same amount of samples and the same reagent strips, both 5 mm wide, were used to make them according to the general method, and the original method was compared with the method of the present invention to detect HCV antibodies.

[0036] Among them, the quality control area was coated with 1 mg / ml SPA (recombinant Staphylococcus aureus protein A), with an amount of 0.5 μl / reagent strip, and the coating was linear. Coat the test area with 0.5 mg / ml of HCV antigen, with an amount of 0.5 μl / reagent strip, and the coating is linear. Chromogen mixture: colloidal gold liquid preparation combined with SPA (volume ratio of 1 mg / ml SPA: colloidal gold is 1:500).

[0037]The samples used in the original method and the method of the present invention are both 5 μl HCV critically positive samples (concentration is about 0.00001 mg / ml), and the colloid...

Embodiment 2

[0039] Embodiment 2 The embodiment of repeatable sample addition

[0040] In this embodiment, continue to take the test of HCV as an example, and its reagent strip, test area, quality control area, chromogenic agent mixture (colloidal gold preparation), reagent strip width, and sample are the same as in Example 1.

[0041] The original method: add 5 μl HCV critical sample (concentration is about 0.00001mg / ml) on the nitrocellulose membrane, after the sample runs through the test area, add 100 μl colloidal gold preparation on the glass fiber. After 1 minute, 5 μl of sample was added, and then every 1 minute, 5 μl of sample was added, and repeated twice, that is, a total of 20 μl of sample was added. A total of 30 minutes to interpret the results.

[0042] The method of the present invention: add 100 μ l of colloidal gold preparation on the glass fiber, wait for the colloidal gold to run out of the glass fiber, enter the sample adding area, add 5 μ l of HCV critical sample (con...

Embodiment 3

[0044] Embodiment 3 Fluorescence test HIV (HIV)

[0045] Using the same amount of samples, the original method and the method of the present invention are compared to detect HIV antibodies at the same time. Among them, the reagent strips are the same, made according to the general method, and the quality control area is coated with 1 mg / ml SPA (recombinant Staphylococcus aureus protein A), with an amount of 0.5 μl / strip. The test area is coated with 0.1 mg / ml HCV antigen, and the dosage is 0.5 μl / strip. The width of the reagent strip is 5mm. Chromogenic reagent mixture: a preparation of fluorescein isothiocyanate (FITC) combined with SPA (volume ratio of 1 mg / ml SPA:FITC is 1:500) was used.

[0046] The original method: Add 5 μl HIV critical sample (concentration is about 0.000005mg / ml) on the nitrocellulose membrane, after the sample runs through the test area, add 100 μl FITC (fluorescein isothiocyanate) preparation on the glass fiber. Timed for 15 minutes, and read the r...

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Abstract

The invention discloses an immunochromatography assay and a device taking a color-developing agent as a sample carrier and capable of repeated sample adding, and the method comprises: 1) preparing a reagent strip according to the conventional method; 2) preparing the color-developing agent according to the conventional method, and mixing the corresponding antigen / antibody of a sample under test with the color-developing agent to form a color-developing agent mixture; 3) adding 50mu l-5ml color-developing agent mixture to a color-developing agent sample adding region; and 4) adding 5mu l-100mu l sample under test to the sample adding region when the color-developing agent enters a sample adding region of a nitrocellulose membrane, and reading the result within 1-30min, wherein, sample adding can be repeated in the meantime. Compared with other conventional methods, in the invention, the color-developing agent enters the nitrocellulose membrane earlier than the sample and has continuous follow-up supply, so the dynamic effects enable the invention to have the advantages of less amount of sample each time, shortened interpretation time, high sensitivity in unit time, clean background of the nitrocellulose membrane after reaction and the like.

Description

technical field [0001] The invention relates to an immunochromatography method, in particular to an immunochromatography method and a device which use a chromogenic agent as a sample carrier and can add samples repeatedly. Background technique [0002] In the past half century, most immunoassays have required expensive analytical equipment and instruments, and professionals have to operate the assays. Although these expensive detection methods have high sensitivity and specificity, they cannot meet the daily needs of people's health protection due to reasons such as expensive, inconvenient to carry, and professionalism. [0003] Colloidal gold immunochromatography (GICA) technology is a simple and rapid immunological detection technology established on the basis of immunofiltration technology in the early 1990s. It was first used by Beggs et al. for human chorionic gonadotropin (HCG) determination. GICA uses nitrocellulose membrane (NC membrane) as the carrier, and uses th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/558G01N33/543G01N21/78
Inventor 姚恺龄陈国治
Owner 益思美诠生物科技(上海)有限公司
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