Colloid selenium test paper for semi-quantitative determination of urinary trace albumin

A urine microalbumin and semi-quantitative detection technology, which is applied in biological testing, material inspection products, and analysis through chemical reactions of materials, can solve problems such as unsuitable for on-site detection, long RIA determination process, and environmental pollution. To achieve the effect of convenient home testing and on-site testing, real-time monitoring and tracking, and simple testing procedures

Active Publication Date: 2013-08-28
LANZHOU UNIVERSITY
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many methods for MAU detection, including radioimmunoassay, enzyme immunoassay and immunoturbidimetric method. However, RIA uses radioisotopes as tracers, which affects the health of operators and seriously pollutes the environment. The instruments used are more expensive, and RIA The determination process is relatively long, which has limitations in clinical application; the cost of chemiluminescence is high, and reagents and instruments are all dependent on imports; ELISA detection requires instruments and is not suitable for on-site detection

Method used

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  • Colloid selenium test paper for semi-quantitative determination of urinary trace albumin
  • Colloid selenium test paper for semi-quantitative determination of urinary trace albumin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Preparation of urinary microalbumin monoclonal antibody

[0022] Mice were immunized with human serum albumin (product of Sigma, USA). The mouse splenocytes whose titer reaches the requirement are selected for hybrid fusion with mouse myeloma cells to prepare hybridoma cells. Select high positive hybridoma cells for multiple subcloning until a cell line that can stably secrete MAU monoclonal antibody is selected.

[0023] Ascites was induced by inoculating hybridoma cells into the peritoneal cavity of mice, and the ascites was extracted and purified to obtain monoclonal antibodies 1 and 2, which proved to target different epitopes on the antigen.

Embodiment 2

[0024] The preparation of embodiment two colloidal selenium

[0025] Add 10% chitosan to the reactor, add ascorbic acid, selenous acid, and ascorbic acid in turn after 2 minutes, and add selenous acid and ascorbic acid at intervals from the fourth minute of the reaction, and then scan the spectrum, and the peaks appear at 500-600nm The maximum absorption peak is appropriate, and the product is collected for future use.

Embodiment 3

[0026] Example 3 Preparation and assembly of colloidal gold test paper for semi-quantitative detection of MAU

[0027] 1. Treatment of the sample liquid absorption part:

[0028] Soak the filter paper or glass fiber paper in 0.1mol / l PBS with pH7.4 for 2min, take it out, dry it at 80°C or dry it by other methods, and then it becomes the absorption part of the sample solution.

[0029] 2. Preparation and Handling of Colloidal Selenium Labeled Sections:

[0030] Mix colloidal selenium and MAU monoclonal antibody 1 in a certain ratio to form MAU monoclonal antibody 1.

[0031] 2.1 Pretreatment of MAU monoclonal antibody 1 to be labeled

[0032] Pre-dialyze MAU monoclonal antibody 1 in 0.005mol / l NaCl, pH7.0 solution at 4°C overnight to remove excess electrolyte, then centrifuge at 6500-13000r / min at 4°C for 60min to remove precipitates, and adjust the protein concentration to 1mg / ml. Can be used for tagging.

[0033] Optimize the experimental conditions for colloidal selenium...

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Abstract

The invention provides a test paper for semi-quantitatively detecting colloid selenium in microdose albumin in urine. The test paper can be applied in the fields of early diagnosis of diabetic nephropathy, hypertensive nephropathy and preeclampsia and injury of kidney caused by various poisonous substances. A plastic bottom plate of the test paper is orderly glued with a sampling liquid absorption part, a colloid selenium labeling part, a detection reaction part and a water absorption part, wherein the colloid selenium labeling part is a single-cloning antibody 1 which is coated by a glass fiber membrane, or acetic acid fiber membrane, or nylon membrane and labeled by the colloid selenium; the detection reaction part is a nitrocellulose membrane which is provided with two detection bands and quality control bands, T1 is a single-cloning antibody 2 which is precisely quantitative, and T2 is the single-cloning antibody 2 with a certain concentration; and the quality control bands are coated by antiplague IgG. The invention provides a method for semi-quantitatively detecting the microdose albumin in a urine sample conveniently, visually and quickly.

Description

1. Technical field [0001] The invention is a colloidal selenium test paper for semi-quantitative detection of urinary microalbumin, through which the test paper can directly and simply determine whether the albumin content in urine is higher than the normal level, and can be applied to early auxiliary clinical diagnosis of diabetic nephropathy. It belongs to the category of in vitro detection equipment for diseases. 2. Background technology [0002] Diabetic nephropathy is a chronic kidney complication of diabetes. Diabetic nephropathy generally refers to diabetic glomerulosclerosis, and its clinical manifestations are proteinuria, edema, hypertension or azotemia, and there are symptoms of increased fasting blood sugar or diabetes. There is no obvious regional difference in this disease, and both type I and type II diabetes can be caused. According to statistics, about 50% of diabetic patients with a disease course of 10-20 years, regardless of age, develop clinical kidney...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/52G01N21/80
Inventor 李红玉张艳徐志良
Owner LANZHOU UNIVERSITY
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