Colloidal gold quick detection test paper of retinal binding protein

A technology for detecting test strips and binding proteins, which is applied in the fields of biomedicine and immunoassay, and can solve the problems of long detection time, shelf life of only half a year, and high cost.

Inactive Publication Date: 2009-11-04
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, the method used to detect RBP in urine is the ELISA method, but the ELISA kit detection will also produce certain false positive results due to the influence of other components in the urine, and the kit needs to be stored at low temperature, and the shelf life is only six months
The detection of urinary RBP is limited to laboratory diagnosis, and it needs to rely on supporting instruments to read data. The cost is relatively high, the detection time is long, and the sampling volume is large. Specialized technicians are required to read and analyze data

Method used

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  • Colloidal gold quick detection test paper of retinal binding protein
  • Colloidal gold quick detection test paper of retinal binding protein
  • Colloidal gold quick detection test paper of retinal binding protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0114] Example 1. Preparation of gold-labeled antibody layer (binding release pad)

[0115] Preparation of colloidal gold

[0116] Using the trisodium citrate method, 0.1% gold chloride was reduced to a colloidal gold solution with 1% trisodium citrate. The quality of the reduced colloidal gold solution is detected by electron microscope observation and 400-700nm wavelength spectrophotometer scanning method, and a colloidal gold solution with a size of about 40nm, uniform size and uniform dispersion is obtained. Adjust the colloidal gold solution to the working concentration (1 mg / ml), and adjust the pH=7.5-8.5.

[0117] Preparation of colloidal gold-labeled anti-urine RBP antibody

[0118] Take 1 ml of colloidal gold solution, adjust the pH to 8.2 with 0.2M potassium carbonate solution, and add 25 μg of commercially available urine RBP antibody (mouse anti-urine RBP monoclonal antibody, purchased from Genetex) or mouse anti-urine RBP prepared by the applicant. Monoclon...

Embodiment 2

[0121] Example 2. Preparation of nitrocellulose membrane with detection line and quality control line

[0122] Get anti-urine RBP polyclonal antibody (prepared according to the method described in 200610117198.7 by this laboratory), adjust its concentration to 1mg / ml (the reagent used to adjust the concentration is PBS), add 2% formaldehyde, and use a film spraying machine (Biodot company, XYZ3050 Model) Spray a test line (t-line, T-line) in the middle of the nitrocellulose membrane, and set the amount of sprayed film to 1 μl / cm.

[0123] Take goat anti-mouse IgG (purchased from SIGMA), adjust the concentration to 2mg / ml (the reagent used to adjust the concentration is PBS), add 2% formaldehyde, and spray it on the middle section of the nitrocellulose membrane with a film sprayer, 0.5cm away from the detection line. For the control line (C-line, C-line), the amount of spray film is set to 1 μl / cm.

[0124] The sprayed film was dried at 37°C for 2 hours, and then blocked with ...

Embodiment 3

[0125] Embodiment 3. Assembly of urine RBP colloidal gold rapid detection test paper

[0126] Assemble the urine RBP colloidal gold rapid detection test strip by the following steps:

[0127] (1) The nitrocellulose membrane with detection line and quality control line obtained in Example 2 is pasted on the PVC base plate;

[0128] (2) Paste the binding release pad prepared in Example 1 on the PVC plate near one end of the test line, so that the binding release pad overlaps with the nitrocellulose membrane at the lower end of the test line by 2 mm;

[0129] (3) Stick water-absorbing glass fiber (sample pad) on the other side of the binding release pad, so that the sample pad overlaps with the binding release pad by 2 mm;

[0130] (4) Paste absorbent paper on the PVC board near one end of the quality control line, so that the absorbent paper overlaps with the nitrocellulose membrane at the top of the quality control line by 2 mm;

[0131] (5) Cut into strips according to the s...

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Abstract

The invention provides colloidal gold quick detection test paper of retinal binding protein (RBP) and an application thereof. The detection test paper comprises a test strip and a baseplate for supporting the test strip, wherein the test strip at least comprises a combining and releasing cushion and a detection cushion; retinal binding protein resistant monoclonal antibodies marked by colloidal gold are wrapped on the combining and releasing cushion in advance; and test lines of retinal binding protein resistant polyclonal antibodies are wrapped on the detection cushion in advance. The RBP detection test paper has low cost and easy preservation, has rapidness, convenience, high sensitivity and good accuracy when used for testing the retinal binding protein, is suitable for clinical and household use and has industrialization.

Description

technical field [0001] The invention relates to the fields of biomedicine and immune detection. More specifically, the present invention relates to a colloidal gold rapid detection test strip for detecting urinary retinol binding protein (RBP). Background technique [0002] Retinol Binding Proteins (RBPs) are a class of carrier proteins responsible for transporting retinol from the liver to various target cells in the human body, and play an important role in the physiological functions of retinol. Seven different RBP subtypes have been found, mainly distributed in the blood circulatory system (RBP4, referred to as RBP), cells (RBP1, RBP2, RBP5, RBP6, RBP7, referred to as CRBPs) and between retinal photoreceptors (RBP3, referred to as IRBP) . [0003] RBP binds to all-trans retinol [Guo Xiaohong, Chu Mingxing, Zhou Zhongxiao, Molecular biology of retinol-binding protein and its gene, Genetics, 26(2), 257-262, 2004], mainly synthesized in the liver . In addition to the li...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/558
Inventor 孙兵董庆犀
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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