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Enzyme-linked detection kit and preparation method thereof

A detection kit and the technology of the kit are applied in the field of biomedicine

Active Publication Date: 2012-04-25
北京健平金星医疗器械有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Houck et al. (J Biol. Chem. 1992, 267:26031) reported an ELISA method for detecting VEGF based on colorimetry, but its detection sensitivity was only ng / ml

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The kit of the present invention can be exemplified by the following substances:

[0023] (1) VEGF receptor and monoclonal antibody are combined to coat the enzyme-linked plate;

[0024] (2) Positive control (VEGF protein freeze-dried powder) 1 tube;

[0025] (3) Negative control (animal serum freeze-dried powder) 1 tube;

[0026] (4) Sample diluent: 0.05% Tween-20, 1% bovine serum albumin, 10mM disodium edetate, 0.05% thimerosal, 0.4M NaCl, pH6.5, 1 bottle;

[0027] (5) Enzyme conjugate: horseradish peroxidase-labeled vascular endothelial growth factor monoclonal antibody, 2 vials;

[0028] (6) Concentrated washing solution: 137mM NaCl, 2.7 mM KCl, 10mM Na2HPO4, 2 mM KH2PO4, 0.05% Tween-20, Ph7.4, 1 bottle;

[0029] (7) Chromogens A and B, of which liquid A is H2O2 solution, and liquid B is 3,3',5,5'-tetramethylbenzidine solution, 1 bottle each;

[0030] (8) The stop solution is 2MH2SO4, 1 bottle;

[0031] (9) 2 sheets of sealing tape.

[0032] The quantities of ...

Embodiment 2

[0090] Embodiment 2: Kit use method

[0091] 1. Take out the required strips from the airtight bag equilibrated to room temperature, and seal the other strips and put them back at 4°C.

[0092] 2. Leave one well as a blank well, and add 100 μl (or 2 drops) of sample diluent to the other wells.

[0093] 3. In addition to the blank well, add 100 μl each of negative control well 1, positive control 2 wells, and serum samples to the corresponding wells, and mix well by flicking. Seal the reaction well with sealing tape and keep at 37°C for 60 minutes.

[0094] 4. Wash the plate 3 times: (1) Automatic plate washer: Shake off the liquid in the well, the washing liquid required to be injected is 350μl, the interval between injection and aspiration is 15-30 seconds, and the plate is washed 3 times. (2) Wash the plate manually: Shake off the liquid in the wells, add 350 μl of washing solution to each well, let it stand for 30 seconds, shake off the liquid, pat dry on thick absorbent ...

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PUM

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Abstract

An enzyme-linked detection kit for detecting VEGF. A kit body contains an enzyme-linked reaction plate coated by a VEGF acceptor and a monoclonal antibody, an enzyme conjugate labelled by HRP and configured by antibody and a protective agent, a freeze-drying powder of the VEGF protein as a positive contrast, a protein freeze-drying powder as a negative contrast, a sample diluent, a PBST concentrated lotion, a chromogenic substrate configured by 3,3',5,5'-tetramethyl benzidine, 2MH2SO4 as a stopping solution and a seal plate gummed paper. The kit can be used in diagnosis and prognosis of cancer, angiopathy, diabetes, retinopathy and rheumatoid arthritis, and has advantages of accuracy, specificity, sensitivity, stability and convenience, etc.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular to an enzyme-linked detection kit for detecting human vascular endothelial cell growth factor (VEGF). [0002] The present invention also relates to the preparation method of the above kit. Background technique [0003] Tumor development and metastasis are multi-stage, complex and highly selective processes involving the intricate relationship between tumor cells and hosts, and the mechanism is still unclear. In recent years, research on the relationship between new angiogenesis and tumor development, metastasis, and prognosis has progressed rapidly, proving that new angiogenesis is a necessary condition for tumorigenesis, metastasis, and the growth of metastatic foci, and vascular endothelial growth factor (VEGF) is the regulator of neovascularization. The most important cytokine produced. Studies have shown that vascular endothelial growth factor (VEGF) has an irreplaceable ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531
Inventor 邹检平
Owner 北京健平金星医疗器械有限公司
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