A kind of test strip and method for detecting urine aβ amyloid

A technology of amyloid protein and test strips, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., and can solve the problems of small target antigens, difficult to reach, high sensitivity and specificity

Active Publication Date: 2021-10-19
HUNAN QIANKANG TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The target antigen is too small, only 4.3kD, and is prone to non-specific binding with other proteins, so it is difficult to achieve high sensitivity and specificity by the commonly used "double antibody sandwich method" or "competition method"

Method used

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  • A kind of test strip and method for detecting urine aβ amyloid
  • A kind of test strip and method for detecting urine aβ amyloid
  • A kind of test strip and method for detecting urine aβ amyloid

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Experimental program
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Embodiment Construction

[0025] 1. Production and purification of anti-Aβ monoclonal antibody

[0026] A commercial mouse hybridoma cell line (BGM02) from Ab-Mart Pty Ltd was purchased to produce mouse anti-Aβ monoclonal antibody. Other commercially available monoclonal antibody clones include 1E8, 4G8, W0-2, 6E10, etc. can also be used alone or in combination. The hybridoma cells were thawed at 45°C using air-permeable hydrophobic cell culture flasks, and 1% glutamine, 1% streptomycin / penicillin, and 20% fetal bovine serum (FBS) were added to the hybridoma cell serum-free medium ( % refers to the mass percentage of the component in the serum-free medium of hybridoma cells).

[0027] When the culture medium turned yellow, the passage cells were carried out, and the ratio of serum in the passage cells decreased from 20%, 10%, 5%, 2% to the serum-free medium (0%), and the cells could survive. This process usually takes a month. Do not aspirate the supernatant until the serum has been consumed.

[00...

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Abstract

A test strip and method for detecting urine Aβ amyloid protein. The test strip for detecting urine Aβ amyloid protein includes a PVC bottom plate (7), and the PVC bottom plate (7) is laid with sequentially overlapped sample pads (1), conjugated binding pads (2), and chromatography pads ( 3) and a water-absorbing pad (4); the conjugated pad (2) is coated with colloidal gold particles conjugated with the monoclonal antibody; the chromatography pad (3) close to the conjugated pad (2) a A detection line (5) is provided on one side, and a control line (6) is provided on the side close to the absorbent pad (4). The detection line (5) is coated with Aβ amyloid-binding polymer, and the control line ( 6) Coated with goat anti-mouse IgG polyclonal antibody. The method has high sensitivity, the detection limit is 40 pg, fast, economical, suitable for routine clinical pathological examination, suitable for general screening of a large number of people and self-screening of home end users, especially for clinically assisting mild cognitive impairment (MCI) Early diagnosis and pre-judgment, and predict whether the patient has a high risk of developing MCI and AD.

Description

technical field [0001] The invention relates to a test strip and a method for detecting urine Aβ amyloid protein. Background technique [0002] Alzheimer's disease (AD) is the most common form of dementia, affecting more than 26 million people worldwide. We and others have found that a large proportion (30%) of cognitively normal older adults exhibit high retention of PET ligands in the brain and low levels of Aβ in the cerebrospinal fluid (CSF) 1-42 , these individuals are currently diagnosed with Alzheimer's disease in the preclinical latent stage. Mild cognitive impairment (MCI) is considered a prodromal stage of AD, and 40-60% of patients meeting MCI criteria will eventually develop clinical AD, accounting for about 5-25% per year. The main etiology of sporadic AD is the accumulation of Aβ and the failure of Aβ clearance. At present, the "amyloid cascade hypothesis" is generally supported in this research field, that is, the metabolic imbalance of Aβ in the brain is t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/58G01N33/577G01N33/558
CPCG01N33/6896G01N33/587G01N33/577G01N33/558G01N2333/4709G01N2800/2821
Inventor 戴正乾顾柏俊
Owner HUNAN QIANKANG TECH CO LTD
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