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Measuring device, kit and method

A technology for measuring devices and kits, which is used in measuring devices, biological tests, and material testing products, etc., can solve the problems of low sensitivity, poor reagent precision, and large amount of antibodies, and achieves low detection cost, stable binding, and specificity. strong effect

Active Publication Date: 2017-11-03
SICHUAN MACCURA BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the disadvantages of this patented technology are: short antigen-antibody binding time, low sensitivity, and poor reagent precision
Defects of this patented technology: low sensitivity and poor reagent precision
Disadvantages of this technology: large amount of antibody, high cost, and low sensitivity
The detection sensitivity requirements of some markers are getting higher and higher. Taking troponin I as an example, the functional sensitivity requirement is below 0.1ng / mL, and the traditional fluorescein can no longer meet the demand.

Method used

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  • Measuring device, kit and method
  • Measuring device, kit and method
  • Measuring device, kit and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Embodiment 1: the preparation of measuring device

[0070] (1) Capture tool (T line) solution preparation at the capture position

[0071] Streptavidin was diluted to 2 mg / mL with 10 mmoL of PB (containing methanol, and the volume fraction of methanol was 3% of the total PB solution volume).

[0072] (2) Preparation of indicator line (C line) solution

[0073] Dilute the goat anti-mouse IgG polyclonal antibody to 1.5 mg / mL with 10 mmoL PB (containing methanol, and the volume fraction of methanol is 3% of the total PB solution volume) solution.

[0074] (3) Coated

[0075] The capture tool solution is coated on the capture position (130) of the nitrocellulose membrane, the indicator line solution is coated on the indicator line (191) of the nitrocellulose membrane, and the volume of the capture tool coated by the capture position (130) is 1 μl / cm.

[0076] (4) drying

[0077] Dry the nitrocellulose membrane prepared in step (3) in a constant temperature oven or a d...

Embodiment 2

[0082] Embodiment 2: the preparation of troponin I kit

[0083] The first component: the first protein labeled with rare earth europium fluorescent microspheres

[0084] Take 1 mg of rare earth europium fluorescent microspheres with a particle size of 200 nm, add EDC until the final concentration of rare earth europium fluorescent microspheres is 0.1 mgEDC / mg fluorescent microspheres, react at room temperature for 30 minutes at 25 degrees, centrifuge to remove free EDC, and then Add the first troponin I monoclonal antibody, centrifuge again, remove the free antibody, block the unbound site with blocking solution, and obtain the first fraction.

[0085] Second component: biotin-labeled second protein

[0086]The second troponin I monoclonal antibody was prepared as a solution A of 1 mg antibody / mL LPBS with PBS, and Biotin-X-X-NHS was prepared as a solution B of 10 mmol Biotin-X-X-NHS / mLDMSO with DMSO. Take 1 mL of solution A and 13.3 μL of solution B and mix evenly, react at...

Embodiment 3

[0091] Embodiment 3: Troponin I sample treatment solution preparation

[0092] The first and second components of the Troponin I kit are mixed to form a treatment solution for testing the Troponin I sample.

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Abstract

The invention discloses an assay device for detecting an analyte in a sample. The device comprises: a placement area for placing a sample; a substrate connected to the placement area so that the sample can be placed along the placement area migration of the substrate; a capture site on the substrate; a capture means present at the capture position, the capture means being avidin or streptavidin. The present invention also discloses a test kit comprising the above measuring device, so as to use the above test kit to determine the presence of the analyte in the sample. The assay device, kit and method of the invention have high sensitivity.

Description

technical field [0001] The present invention relates to methods and devices for detecting the presence of analytes. Background technique [0002] The current rapid immunochromatographic detection card mostly uses colloidal gold, colored latex particles or fluorescein as markers. Rapid detection products developed based on colloidal gold labeling technology have low sensitivity and large batch-to-batch differences, and can only be qualitative or semi-quantitative. Although the difference between batches of colored latex particles has been improved, the sensitivity is still low, and it can only be qualitative or semi-quantitative. [0003] The sensitivity of immunochromatography based on fluorescein labeling technology has been greatly improved, and quantitative detection can also be performed, but because of the small STOCK shift and short fluorescence lifetime, the detection sensitivity is easily affected by background fluorescence. [0004] CN102192983A discloses a time-r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/533
Inventor 刘东泽赵书阁马应霞方袁梦梦
Owner SICHUAN MACCURA BIOTECH CO LTD