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A kit and method for time-resolved fluorescent quantitative detection of urinary microalbumin

A time-resolved fluorescence and urine microalbumin technology, applied in fluorescence/phosphorescence, measurement devices, analytical materials, etc., can solve the problems of time-consuming instruments, low sensitivity, and high sensitivity of enzyme-linked immunosorbent assays, and achieve stable and stable labeling products. Avoid fluorescence interference and detect fast and easy results

Active Publication Date: 2019-12-17
厦门奥德生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the colloidal gold method has the advantages of simple and fast operation, but its sensitivity is low and its quantification is inaccurate; the enzyme-linked immunoassay method has high sensitivity and large sample volume can be quantitatively detected, but the operation time is long and the automation is low; immunoturbidimetric method and chemiluminescence method are relatively Sensitive and accurate, it can be applied to automatic biochemical analyzers, but requires instruments and takes a long time, is suitable for processing a large number of samples, and cannot meet the purpose of rapid detection; immunofluorescence method is to covalently bind urine microalbumin antibodies to fluorescent microspheres On the surface active group, the result is judged by whether the detection line generates fluorescence after excitation. It is fast, convenient, accurate and quantitative, and has the advantages of high sensitivity and stable markers. It is widely used in the field of medical immunoassay

Method used

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  • A kit and method for time-resolved fluorescent quantitative detection of urinary microalbumin
  • A kit and method for time-resolved fluorescent quantitative detection of urinary microalbumin
  • A kit and method for time-resolved fluorescent quantitative detection of urinary microalbumin

Examples

Experimental program
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Effect test

Embodiment 1

[0041] Example 1: Preparation of time-resolved fluorescence quantitative detection kit for urinary microalbumin

[0042] The time-resolved fluorescence quantitative detection kit for microalbumin in urine uses the principle of competitive immunochromatography to detect microalbumin in human urine. Consists of time-resolved fluorescent test paper card, sample diluent and SD card containing calibration curve.

[0043] Such as figure 1 As shown, in this embodiment, the sample pad 1, the coating film 2 and the absorbent paper 3 are sequentially overlapped on the substrate 7, the sample pad 1 is a sample application area, and is used to absorb the urine test sample to be tested, and the coating film 2 There are microsphere line, detection line and quality control line on it.

[0044] Rare earth europium fluorescent microspheres (about 200 nm in diameter, 1% europium solid content) with specific excitation light (365nm) / emission light (615nm) wavelengths were used on the microsphe...

Embodiment 2

[0056] Example 2: Quantitative detection of time-resolved fluorescence quantitative detection kit for urinary microalbumin

[0057] 1. Drawing of standard curve

[0058] The time-resolved fluorescent test paper card prepared according to Example 1 was added with different concentrations of albumin antigen quality controls (302.5mg / L, 247.5mg / L, 192.5mg / L, 137.5mg / L, 82.5mg / L, 38.5mg / L L, 11.0mg / L, and 2.0mg / L have a total of eight concentrations, and each concentration has three repetitions, all of which are obtained by diluting 10mg / ml human albumin recombinant antigen with 0.85% normal saline), adding sample diluent, and chromatographically After 10 minutes, read the C and T line fluorescence signals and C / T value through the AFS-1000 dry-type fluorescence immunoassay analyzer produced by Guangzhou Lanbo Biotechnology Co., Ltd. The experimental results and analysis are shown in Table 1:

[0059]

[0060] Take the LOG value of the trace albumin control product concentrati...

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Abstract

The invention relates to a kit and a method for quantitatively detecting urine microalbumin through time-resolved fluorescence. The kit comprises a time-resolved fluorescence test paper card, sample diluent and a DS card containing a calibration curve, wherein the time-resolved fluorescence test paper card comprises a detection test paper strip which consists of a bottom liner, and a sample pad, a nitrocellulose membrane and absorbent paper which are pasted on the bottom liner; the nitrocellulose membrane is successively provided with a microsphere line, a detection line and a quality control line; the microsphere line consists of time-resolved fluorescence microspheres marked with human seroalbumin monoclonal antibodies; the detection line is coated with human seroalbumin recombinant antigens; the quality control line is coated with goat anti-mouse IgG. The kit provided by the invention can accurately and quantitatively detect the content of microalbumin in human urine; markers are used for connecting the microspheres with antibodies through covalent bonds; and the marked products are stable. The kit has the characteristics of wide detection range (2 to 300mg / ml), high sensitivity (the detection limit is 2300mg / ml), high accuracy, fast and easy detection and the like.

Description

[0001] (1) Technical field [0002] The invention relates to a kit and a detection method for time-resolved fluorescence quantitative detection of urinary microalbumin. [0003] (2) Background technology [0004] Microalbuminuria refers to the presence of microalbumin in the urine. Albumin is a normal protein in the blood. Under normal human metabolism, there is very little albumin in the urine, and the albumin per liter of urine does not exceed 20mg (<20mg / L), so it is called microalbumin. However, when diabetic nephropathy, hypertensive nephropathy and other early kidney damage and cardiovascular disease complications occur, the intrinsic cell damage of the kidney will be caused, and the structure and function of the intrinsic cell of the kidney will change, resulting in the increase of albumin content in the urine. The microalbuminuria in the urine is in the range of 20-200mg / L, which belongs to microalbuminuria. When the microalbuminin in the urine exceeds 200mg / L, it p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64G01N33/577
CPCG01N21/6408G01N33/577
Inventor 张云张肖杨福太
Owner 厦门奥德生物科技有限公司
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