Homogeneous chemiluminiscence detection kit and application thereof

A homogeneous chemiluminescence, detection kit technology

Pending Publication Date: 2020-02-21
BEYOND DIAGNOSTICS (SHANGHAI) CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, with the progress of the detection industry, there is an increasing demand for ultrasensitive reagents, which not only require extremely high sensitivity, but also require a very wide linear range. It is difficult for existing kits to meet the above detection conditions.

Method used

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  • Homogeneous chemiluminiscence detection kit and application thereof
  • Homogeneous chemiluminiscence detection kit and application thereof
  • Homogeneous chemiluminiscence detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0157] Embodiment 1: Preparation of acceptor reagent

[0158] (1) Preparation and characterization process of aldehyde-based polystyrene latex microspheres

[0159] 1. Prepare a 100ml three-necked flask, add 40mmol styrene, 5mmol acrolein, and 10ml water, stir for 10min, and pass N 2 30min.

[0160] 2. Weigh 0.11g of ammonium persulfate and 0.2g of sodium chloride, dissolve them in 40ml of water to form an aqueous solution. This aqueous solution is added in the reaction system of step 1, continues to pass through N 2 30min.

[0161] 3. The temperature of the reaction system was raised to 70° C. and reacted for 15 hours.

[0162] 4. Cool the emulsion after the reaction to room temperature and filter it with a suitable filter cloth. The obtained emulsion was washed with deionized water for several times of centrifugation and sedimentation until the conductivity of the supernatant at the beginning of centrifugation was close to that of deionized water, then diluted with wa...

Embodiment 2

[0176] Embodiment 2: the determination of kit sensitivity and upper limit of detection

[0177] Define the sensitivity point as when the signal of concentration 2C0 is higher than the signal of twice the concentration of C0, that is, RLU(2C0)>2RLU(C0), then the sensitivity of the corresponding detection reagent is C0. The upper limit of detection is defined as the corresponding concentration obtained by substituting the detection signal with a concentration of 1000 ng / ml into the curve of concentration and signal.

[0178] (1) Dilute the cTnI antigen to 5pg / ml, 10pg / ml, 20pg / ml, 30pg / ml, 40pg / ml, 50pg / ml, 100pg / ml, 1000pg / ml, 5000pg / ml, 10000pg / ml, 50000pg / ml ml, 1000ng / ml series concentration, will adopt the same method as Example 1 to prepare different particle diameters (50nm, 80nm, 110nm, 140nm, 170nm, 200nm, 250nm, 300nm, 350nm, 400nm) coated with cTnI monoclonal antibody 1 Acceptor reagent (concentration is 100ug / ml), then with the same biotin-labeled cTnI monoclonal an...

Embodiment 3

[0189] Embodiment 3: Preparation of acceptor reagent

[0190] (1) Preparation of antibody-coupled receptor microspheres with an average particle size of about 250 nm

[0191] 1.1 Preparation and characterization process of aldehyde-based polystyrene latex microspheres

[0192] 1) Prepare a 100ml three-neck flask, add 40mmol styrene, 5mmol acrolein, and 10ml water, stir for 10min, and pass N 2 30min;

[0193] 2) Weigh 0.11g of ammonium persulfate and 0.2g of sodium chloride, dissolve them in 40ml of water to form an aqueous solution. This aqueous solution is added in the reaction system of step 1, continues to pass through N 2 30min;

[0194] 3) The temperature of the reaction system was raised to 70° C., and the reaction was carried out for 15 hours;

[0195] 4) Cool the emulsion after the reaction to room temperature, and filter with a suitable filter cloth. The obtained emulsion is washed with deionized water by centrifugation and sedimentation until the conductivity...

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Abstract

The invention relates to a homogeneous chemiluminiscence detection kit and application thereof and belongs to the technical field of chemiluminiscence. The kit comprises an acceptor reagent and a donor reagent; the acceptor reagent comprises at least two acceptor microspheres with different particle sizes; and the acceptor microspheres can react with singlet oxygen active oxygen to generate detectable chemiluminiscence signals; and the donor reagent comprises donor microspheres which can generate singlet oxygen active oxygen under an excited state. The kit disclosed by the invention not only has ultrahigh sensitivity, but also has a very wide detection range. Besides, the reaction speed of the small-particle-size light-emitting microspheres is high, and thus, with the immunoassay method adopted for detection, detection time can be shortened, and reaction speed is increased.

Description

technical field [0001] The invention belongs to the technical field of chemiluminescence, and in particular relates to a homogeneous chemiluminescence detection kit and an application thereof. Background technique [0002] Chemiluminescence analysis is a detection method using light waves emitted by chemiluminescent substances. Chemiluminescent substances are used as labels in nucleic acid detection and immunodetection. For example, a certain molecule in a specific binding pair can be combined with a luminescent substance in various ways to form a luminescent microsphere composition. The microsphere composition can react with the detected substance (another molecule in the specific binding pair) in the sample, and is distributed in the solid phase and the liquid phase, and the distribution ratio is related to the amount of the detected substance. By measuring the amount of luminescence in the solid phase or liquid phase, the corresponding concentration of the detection sub...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76
CPCG01N21/76
Inventor 杨阳康蔡俊赵卫国刘宇卉李临
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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