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Detection kit and detection method for soluble urokinase-type plasminogen activator receptor

A detection kit, plasminogen technology, applied in measurement devices, biological material analysis, instruments, etc., can solve the problems of high specificity, low detection sensitivity and accuracy of ELISA, and improve detection sensitivity and specificity. performance, optimized response time, simple operation

Inactive Publication Date: 2018-06-29
无锡壹闪生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the ELISA method is mostly used in clinics to detect the content of suPAR. However, the heterogeneous reaction of ELISA has the disadvantages of high background and poor specificity, low detection sensitivity and accuracy, and has great limitations.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0024] 1. Preparation of Magnetic Separation Reagents

[0025] 1. Preparation of magnetic particle buffer

[0026](1) Weigh TRIS 4.58g / L, NaCl 6.81g / L, TRITON X-100 0.5mL / L in a 1L container; (2) Measure Proclin-300 0.5-1mL in a 1L container; measure 800mL for purification Put water in the above container and stir thoroughly; (3) adjust the pH value to 7.95-8.05; (4) weigh 3-5g of BSA into the above 1L container, and wait until it is fully dissolved; (5) finally make it to 1000mL, 0.22 um microporous filter; mark it well and store it in a cold storage at 2-8°C for later use.

[0027] 2. Preparation of magnetic separation reagents

[0028] (1) Dissolve 1.0 mg disuccinimidyl suberate in 50 μL DMSO to obtain a disuccinimidyl suberate solution; dissolve 2 mg streptavidin in 0.1 mol / 1 mL of phosphate buffered saline to a total volume of 1 mL to obtain a streptavidin solution;

[0029] (2) Determine the input amount of disuccinimidyl suberate, draw the disuccinimidyl suberate s...

Embodiment 1

[0070] This embodiment provides a soluble urokinase-type plasminogen activator receptor detection kit, including: magnetic separation reagents, antibody reagents, calibrator, quality control products, concentrated cleaning solution and substrate solution; wherein, the antibody reagent Containing alkaline phosphatase-labeled antibody and biotin-labeled antibody, the magnetic separation reagent includes magnetic particles coated with streptavidin.

[0071] 1. Preparation of Magnetic Separation Reagents

[0072] 1. Preparation of magnetic particle buffer

[0073] (1) Weigh TRIS 4.58g / L, NaCl 6.81g / L, TRITON X-100 0.5mL / L in a 1L container; (2) Measure Proclin-300 0.5mL in a 1L container; measure 800mL purified water (3) Adjust the pH value to 8.00; (4) Weigh 4g of BSA into the above 1L container and wait for it to fully dissolve; (5) Finally, set the volume to 1000mL and filter through a 0.22um microporous filter ; Mark it and store it in a cold room at 2-8°C for later use.

...

Embodiment 2

[0105] This embodiment provides a method for detecting soluble urokinase-type plasminogen activator receptor (suPAR) using a soluble urokinase-type plasminogen activator receptor detection kit, comprising steps:

[0106] S1: Add 50 μL standard substance, 50 μL quality control substance and 50 μL serum of the sample to be tested to the bottom of the chemiluminescence reaction tube;

[0107] S2: Add 50 μL of antibody reagent to the bottom of the chemiluminescence reaction tube;

[0108] S3: Gently shake the reaction tube for 30 seconds with a multi-functional homogeneous mixer, and then place it in a thermostat at 37°C for 15 minutes;

[0109] S4: Add 50 μL of magnetic separation reagent to the bottom of the chemiluminescence reaction tube;

[0110] S5: Repeat step S3 once;

[0111] S6: Put the reaction tubes on the magnetic separator, make sure that the bottom of each tube is in contact with the surface of the separator, and settle for 2 minutes; then slowly invert the separa...

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Abstract

The invention relates to a detection kit and a detection method for a soluble urokinase-type plasminogen activator receptor. The detection kit comprises a magnetic separation reagent, an antibody reagent, a calibration product, a quality control product, a concentrated cleaning solution and a substrate solution. A quantitative detection kit for the soluble urokinase-type plasminogen activator receptor (suPAR), which is provided by the invention, adopts a method for combining chemiluminescence immunoassay with a magnetic separation technique, and is used for greatly improving the detection sensitivity and specificity of an allergen-specific antibody; the detection result is enabled to be truer and more reliable; meanwhile, the reaction time and reaction steps are optimized, so that the operation is enabled to be simpler.

Description

technical field [0001] The invention relates to a soluble urokinase-type plasminogen activator receptor detection kit and a detection method, which are used for quantitative determination of the content of the soluble urokinase-type plasminogen activator receptor in human serum in vitro. Background technique [0002] The urokinase-plasminogen activator system consists of soluble urokinase-type plasminogen activator, soluble urokinase-type plasminogen activator receptor (suPAR) and its inhibitors. This system participates in cell peripheral functions, cell migration, angiogenesis, tissue remodeling, etc. through proteolysis, signal transduction, and activation of activating factors. Soluble urokinase-type plasminogen activator receptor (suPAR) is the soluble form of urokinase-type plasminogen activator receptor, under stimuli such as inflammation, soluble urokinase-type plasminogen activator receptor (suPAR) is cleaved from the cell surface and released into body fluids, inc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/543
CPCG01N33/54326G01N33/57484G01N2800/7095
Inventor 奚伟红
Owner 无锡壹闪生物科技有限公司