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Preparation and application of immunochromatography method and test paper for quantitative detection of prealbumin

A technology for quantitative detection of prealbumin, applied in measurement devices, analytical materials, instruments, etc., can solve the problems of time-consuming, unsuitable for rapid detection, poor precision and accuracy, etc.

Inactive Publication Date: 2017-05-17
北京中生金域诊断技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Most of these methods are time-consuming, and the samples need to be pre-treated, special equipment is required, and there are disadvantages such as poor precision and accuracy, and they are not suitable for clinical on-site rapid detection

Method used

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  • Preparation and application of immunochromatography method and test paper for quantitative detection of prealbumin
  • Preparation and application of immunochromatography method and test paper for quantitative detection of prealbumin
  • Preparation and application of immunochromatography method and test paper for quantitative detection of prealbumin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Preparation of prealbumin fluorescent microsphere immunochromatographic test strips in quantitative detection samples

[0064] In this example, the bottom plate material is PVC plastic sheet, which is purchased from Yuyao Fucheng Plastic Chemical Co., Ltd.; the material of the sample pad is glass fiber film, which is purchased from Shanghai Jinbiao Biotechnology Co., Ltd.; the material of the bonding pad is glass The fiber membrane was purchased from Shanghai Jinbiao Biotechnology Co., Ltd.

[0065] 1. Preparation of nitrocellulose membrane

[0066] ① Screen suitable anti-prealbumin antibody and donkey anti-mouse antibody;

[0067] ②Preparation of detection line and quality control line on nitrocellulose membrane;

[0068] Use 0.01M PB buffer (with 3% sucrose) at pH 7.2 to 7.4 to adjust the concentration of anti-prealbumin antibody II to 0.6 mg / mL, and the concentration of donkey anti-mouse antibody to 1.0 mg / mL, using Bio-Dot XYZ3050 The three-dimensional ...

Embodiment 2

[0077] Quantitative detection of prealbumin in the sample of embodiment 2

[0078] The test strip prepared in Example 1 is used to detect prealbumin in the sample, and the steps include:

[0079] (1) Add solutions containing prealbumin standards with different known concentrations (0, 2.5, 5.0, 12.5, 25.0, 37.5, 75.0, 100, 200ug / L) into the sample hole of the test strip, After reacting for 15 minutes, place the test strip under the excitation light source with a wavelength of 470nm, measure the fluorescence intensity of the strips on the detection line and the quality control line, and take the ratio of the fluorescence intensity of the strips on the detection line and the quality control line as the ordinate, the former The concentration of albumin is drawn as the standard curve on the abscissa (such as image 3 shown). The results show that the present invention has a good linear correlation (R 2 =0.998), and its detection sensitivity was 2.5 μg / L.

[0080] (2) Add the a...

Embodiment 3

[0081] Quantitative detection of prealbumin in the sample of embodiment 3

[0082] Take the test strip prepared in Example 1, open the aluminum foil bag, take out the test strip, lay it flat on the table, take 10 μL of blood sample into the reagent bottle filled with 2mL 0.01M pH 7.4 phosphate buffer solution, mix thoroughly and take 100 μL Add it into the sample hole of the test strip, react for 15 minutes, place the test strip under the excitation light source, measure the fluorescence intensity of the strips on the detection line and the quality control line, and compare the fluorescence intensity of the detection line with the fluorescence intensity of the quality control line. The ratio of the standard curve in Example 2 can be obtained to detect the content of prealbumin in the blood sample.

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Abstract

The invention provides preparation and application of an immunochromatography method and test paper for quantitative detection of prealbumin. The test paper consists of a bottom plate, a sample pad, a conjugate pad, a nitrocellulose membrane and a piece of water absorbing paper, wherein the sample pad, the conjugate pad, the nitrocellulose membrane and the water absorbing paper are sequentially adhered to the bottom plate in a lap joint manner and are respectively and partially overlapped; a detection line and a quality control line are arranged on the nitrocellulose membrane; the quality control line is wrapped with a donkey-anti-rat antibody. A method comprises the following steps: (1) adding a solution which is of known concentration and comprises a prealbumin standard substance onto the sample pad of the test paper, and drawing a standard curve; (2) adding a solution of a substance to be tested, performing reaction, and reading content according to standard curves. The test paper and the detection method provided by the invention are high in sensitivity, that is, up to pg / mL, wide in detection linearity, convenient to operate, and short in time, that is, results can be obtained within only 10-20 minutes.

Description

technical field [0001] The invention belongs to the technical field of medical immunological in vitro diagnosis, and in particular relates to an immunochromatographic method for quantitatively detecting prealbumin, preparation and application of a test strip. Background technique [0002] Prealbumin is a serum protein synthesized by liver cells, which migrates before albumin during electrophoresis, hence the name prealbumin. Prealbumin is a tetramer composed of 4 identical subunits, with a molecular weight of about 54980 and an extinction coefficient (E280nm) of 13.6. Its half-life is very short, about 1.9 days. Prealbumin participates in serum thyroxine and retinol transport, and has thymus hormone activity, can increase the body's immunity by promoting the maturation of lymphocytes. [0003] Prealbumin in the blood is a negative acute phase protein whose concentration level decreases with inflammation and malignant disease. The half-life of prealbumin is very short, whic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558
CPCG01N33/558
Inventor 杨亚杰李超辉王加义凌燕王维赖卫华熊勇华
Owner 北京中生金域诊断技术股份有限公司
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