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Fluorescence immunoassay quantitative detection kit of microalbuminuria, and preparation method thereof

A technology of microalbumin and fluorescent immunoassay, which is applied in the field of microalbumin fluorescent immunoassay quantitative detection kit and its preparation, can solve the problems of low accuracy, limited reliability, poor stability, etc., and achieve high sensitivity and high accuracy , the effect of easy operation

Inactive Publication Date: 2013-01-16
GETEIN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method is simple, the accuracy is not high, and it cannot be quantitatively detected
Chinese patent application No. 01126895.6 provides a urine albumin chip for rapid detection of kidney damage, but it does not elaborate on the important characteristics of the chip, such as the preparation process, detection sensitivity, and minimum detection amount, which limits the reliability of its application
[0005] (1) The colloidal gold labeling process is an electrostatic adsorption process, which is a physical process, so the stability in the liquid phase is poor, which often causes the labeled protein molecules to fall off again;
[0006] ⑵ Only when the gold particles gather to a certain amount, the human naked eye can observe the purple-red band, and the contrast between the color and the background is not large, thus limiting the detection sensitivity

Method used

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  • Fluorescence immunoassay quantitative detection kit of microalbuminuria, and preparation method thereof
  • Fluorescence immunoassay quantitative detection kit of microalbuminuria, and preparation method thereof
  • Fluorescence immunoassay quantitative detection kit of microalbuminuria, and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 The preparation method of microalbumin fluorescent immunoassay kit is as follows:

[0034] 1) Preparation of fluorescent latex microspheres

[0035] Preparation of fluorescent latex microspheres: Dilute latex microspheres with a particle size of 153 nm to a final concentration of 20 mg / ml with an adsorption buffer (50 mM, citrate buffer at pH 5.8) to obtain a latex microsphere suspension ; add an appropriate amount of fluorescein Cy5-labeled streptavidin (purchased from Shanghai Yanjing Biotechnology Co., Ltd.) in the adsorption buffer, the final volume is 5ml; the above-mentioned latex microsphere suspension is added to the above-mentioned containing fluorescein-labeled chain In the adsorption buffer of mycoavidin, a mixed solution was prepared; the resulting mixed solution was incubated at room temperature for 1-2 hours, and stirred continuously, and then centrifuged to collect the precipitate, and the precipitate was stored in a buffer solution (containing ...

Embodiment 2

[0050] Example 2 The preparation method of another microalbumin fluorescence immunoquantitative detection kit is as follows:

[0051] 1) Preparation of fluorescent latex microspheres

[0052] Preparation of fluorescent latex microspheres: Dilute latex microspheres with a particle size of 500 nm to a final concentration of 30 mg / ml with an adsorption buffer (50 mM, citrate buffer at pH 5.8) to obtain a latex microsphere suspension ; add an appropriate amount of red fluorescein rhodamine-labeled streptavidin (purchased from Shanghai Enzyme Biotechnology Co., Ltd.) in the adsorption buffer, the final volume is 6ml; the above-mentioned latex microsphere suspension is added to the above-mentioned containing red fluorescent In the adsorption buffer of sulhodamine-labeled streptavidin, a mixed solution was prepared; the resulting mixed solution was incubated at room temperature for 1-2 hours, and kept stirring, then centrifuged, and the precipitate was collected, and the precipitate ...

Embodiment 3

[0059] Example 3 Microalbumin Fluorescence Immunoquantitative Assay Test Strip Quantitative Detection

[0060] 1) Draw a standard curve

[0061] Add different concentrations of albumin antigen standard substance (take six different concentrations, respectively 10, 20, 40, 80, 160, 200 mg / L) on the microalbumin fluorescent immunoquantitative assay test strip sample pad prepared by Example 1 , with 3 replicates for each concentration), the signals of detection line 7 and quality control line 8 were read by the fluorescence immunoquantitative analyzer Getein1100 of Nanjing Gedan Biotechnology Co., Ltd. The experimental results and analysis are shown in Table 1.

[0062] Table 1 Albumin Standard Test Results

[0063]

[0064] Draw a standard curve with the concentration of the microalbumin antigen standard substance and the average value of the determined signal, such as figure 2 shown.

[0065] 2) Detection of linear range

[0066] The test strip prepared in Example 1 of ...

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Abstract

The invention relates to a fluorescence immunoassay quantitative detection kit of microalbuminuria, and a preparation method thereof. A sample pad, a combination pad, a nitrocellulose membrane and absorbent paper are pasted on the bottom liner of the quantitative detection kit in a successively lapped manner, wherein the combination pad is coated by fluorescent latex microsphere-marked human albumin antibodies; and the nitrocellulose membrane is coated by a detection line of albumin recombinant antigens and a quality control line of rabbit anti-mouse IgG antibodies. The fluorescence immunoassay quantitative detection kit can detect the content of microalbuminuria in human urine accurately and quantitatively, and has the characteristics of simple operation, high accuracy, high sensitivity, low cost, etc.

Description

technical field [0001] The invention belongs to the field of medical immunological in vitro diagnosis, and in particular relates to a microalbumin fluorescent immunological quantitative detection kit and a preparation method thereof. Background technique [0002] Under normal circumstances, when albumin in plasma passes through the glomerular capillary network, it seldom enters the primary urine. Even if a small amount passes through, 99% of it can be reabsorbed by the convoluted renal tubules, so albumin does not exist in the final urine. However, due to factors such as renal failure, diabetes and cardiovascular disease complications, albumin can be pathologically elevated. In general, urinary protein is significantly increased, which is common in severe glomerular lesions, and a mild increase indicates renal tubular damage. Albumin in human urine reaches a concentration of 30-300mg / day or 20-200mg / L, which is called microalbumin (also known as microalbuminuria, Micoralbum...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/533
Inventor 苏恩本王勇陈伟黄力
Owner GETEIN BIOTECH
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