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Chemiluminescence analysis method and application thereof

A chemiluminescence and analytical method technology, applied in chemiluminescence/bioluminescence, material excitation analysis, and analysis by chemical reaction of materials, etc. problems, to achieve the effect of improving detection sensitivity, shortening detection time, and improving detection performance

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

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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 the existing chemiluminescence analysis methods to meet the above detection conditions.

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  • Chemiluminescence analysis method and application thereof
  • Chemiluminescence analysis method and application thereof
  • Chemiluminescence analysis method and application thereof

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Embodiment approach

[0092] The present invention will be described in more detail below.

[0093] The first aspect of the present invention relates to a chemiluminescence analysis method, which analyzes and judges whether there Contains the target molecule to be tested and / or the concentration of the target molecule to be tested.

[0094] In some embodiments of the present invention, the method analyzes and judges whether the test sample contains the test sample by detecting the intensity of the chemiluminescent signal generated by the reaction of two kinds of acceptor microspheres with different particle sizes and active oxygen in the test sample. The concentration of target molecule and / or target molecule to be tested.

[0095] In some embodiments of the present invention, the particle size difference between the two acceptor microspheres with different particle sizes is not less than 100 nm; preferably not less than 150 nm; more preferably not less than 200 nm. In some specific embodiments o...

Embodiment 1

[0154] Example 1: Preparation of acceptor microspheres of different particle sizes

[0155] (1) Preparation and characterization process of aldehyde-based polystyrene latex microspheres

[0156] 1. Prepare a 100ml three-necked flask, add 40mmol styrene, 5mmol acrolein, and 10ml water, stir for 10min, and pass N 2 30min.

[0157] 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.

[0158] 3. The temperature of the reaction system was raised to 70° C. and reacted for 15 hours.

[0159] 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 deioniz...

Embodiment 2

[0173] Example 2: Determination of the Sensitivity and Detection Limit of the Chemiluminescence Analysis Method Using Acceptor Microspheres with Different Particle Sizes

[0174] 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.

[0175] (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 The acceptor mi...

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Abstract

The invention relates to a chemiluminiscence analysis method and belongs to the technical field of chemiluminiscence analysis. According to the chemiluminiscence analysis method, the intensity of chemiluminiscence signals generated by the reaction of at least two receptor microspheres with different particle sizes in a to-be-detected sample and active oxygen is detected, so that whether the to-be-detected sample contains to-be-detected target molecules and / or the concentration of the to-be-detected target molecules can be analyzed and judged. The method provided by the invention not only has ultrahigh sensitivity, but also has a very wide detection range. Besides, the reaction speed of small-particle-size light-emitting microspheres is high, and thus, with the immunoassay method adopted, detection time can be shortened, and reaction speed is increased.

Description

technical field [0001] The invention belongs to the technical field of chemiluminescence analysis, and in particular relates to a chemiluminescence analysis method 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 subs...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N21/63G01N33/543
CPCG01N21/76G01N33/54346G01N21/63G01N33/543
Inventor 杨阳康蔡俊赵卫国刘宇卉李临
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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