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A kit for rapid detection of upa and pai-1

A technology of PAI-1 and reagent kit, which is applied in the direction of measuring devices, microbial determination/inspection, instruments, etc., can solve the problems of no simultaneous detection, etc., and achieve the effects of reducing contamination opportunities, high specificity, and rapid assessment of prognostic symptoms

Inactive Publication Date: 2019-02-12
百创汇国际生物科技(武汉)股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no relevant research report on the simultaneous detection of uPA and PAI-1 by immunochromatography

Method used

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  • A kit for rapid detection of upa and pai-1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The preparation of embodiment 1 fluorescent latex particles

[0049] Preparation:

[0050] Take 10g of methyl methacrylate, 0.02g of rhodamine B, 0.01g of sodium 2-methallylsulfonate and 100ml of ultrapure water in a reaction kettle, stir and heat at a stirring speed of 200rpm, and feed nitrogen , when the temperature of the system rises to 70°C, add 4ml of potassium persulfate, keep it warm for 120 minutes, cool down after the reaction, use a syringe filter to adjust the particle size of the reaction product to 100nm, then filter the reaction product, centrifuge to remove the supernatant, precipitate Wash 3 times to get fluorescent latex particles,

Embodiment 2

[0051] Example 2 Preparation of fluorescent latex particle-labeled antibody

[0052] 1. Preparation of uPA antibody labeled with fluorescent latex particles

[0053] Preparation:

[0054] (1) Centrifuge the fluorescent latex microsphere solution at 20000rpm for 10min, remove the supernatant and collect the precipitate, then add PBS buffer solution to adjust the concentration of fluorescent latex microsphere to 1.0╳10 12 pieces / mL, dispersed in 100W ultrasonic wave for 30s to obtain latex microsphere dispersion;

[0055] (2) Add 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxyl sulfosuccinimide successively in latex microsphere dispersion liquid, described 1-ethyl -The final concentration of 3-(3-dimethylaminopropyl) carbodiimide in latex microsphere dispersion is 6mg / mL, the final concentration of N-hydroxyl sulfosuccinimide in latex microsphere dispersion The concentration is 4mg / mL, shake and mix evenly, place in a constant temperature shaker for 30min, then c...

Embodiment 3

[0059] Embodiment 3 Preparation of detection kit of the present invention

[0060] 1. Sample preparation:

[0061] (1) Preparation of fluorescent latex-labeled stable solution: Take the preparation of 100mL fluorescent latex-labeled stable solution as an example, as follows:

[0062] Take respectively 50 mL of the fluorescent latex particle-labeled uPA antibody solution and the fluorescent latex particle-labeled PAI-1 antibody solution prepared in Example 2, mix well, add 6 g of 3-(N-morpholine) propanesulfonic acid, 1.5 g of Sodium dialkyl sulfonate, 0.5 g sodium alginate, and 2 g glycine are mixed evenly to obtain a fluorescent latex labeling stable solution.

[0063] (2) Preparation of uPA antibody solution or PAI-1 antibody solution or goat anti-mouse immunoglobulin IgG solution: take 1 mg of uPA antibody or PAI-1 antibody or goat anti-mouse immunoglobulin IgG, add 1ml PBS buffer solution, dissolve, Instantly.

[0064] (3) Preparation of sample pad pretreatment solution...

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Abstract

The invention relates to a kit for rapidly detecting uPA and PAI-1. The kit comprises a detection reaction carrier, a uPA standard substance and a PAI-1 standard substance, the detection reaction carrier is a test strip coated with a film material, and the test strip comprises a PVC baseboard, and a sample pad, a fluorescent marker binding pad, a nitrocellulose membrane and an absorbent paper which are sequentially arranged on the surface of the PVC baseboard from a sample application end. The kit can simultaneously quantitatively detects the uPA and the PAI-1 in a sample, can detect the content of the uPA and the PAI-1 through one-time sample addition, greatly simplifies the operating process, and has high detection sensitivity, the lowest uPA detection limit is 0.01 ng / ml, the lowest PAI-1 detection limit is 0.1 ng / ml, and the kit also has the advantages of no cross reaction in the detection process of the uPA and the PAI-1, no interference and high specificity, and can be used to rapidly assess the prognosis symptoms of tumor patients.

Description

technical field [0001] The invention belongs to the field of medical biological immunological detection reagents, in particular to a kit for rapid detection of uPA and PAI-1. Background technique [0002] Malignant tumors seriously threaten the life safety and physical and mental health of the broad masses of the people. Early detection is widely considered the best way to reduce cancer mortality. At present, a variety of special examinations are used clinically in order to improve the early diagnosis rate of tumors, including X-ray examination, ultrasonography, CT examination, magnetic resonance examination, endoscopy and so on. These examination methods can diagnose tumors from different angles and sides, and each has its own limitations. The emergence of tumor markers has given people great hope for the early diagnosis of tumors. The detection of tumor markers is of great significance for auxiliary diagnosis of tumors, judgment of tumor prognosis, outcome, and evaluati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/37
CPCG01N33/57484G01N33/6893G01N2333/8132
Inventor 陈立国邹伟权张亚丽李庆祥母润红王涛苏焱
Owner 百创汇国际生物科技(武汉)股份有限公司