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Method for qualitatively and quantitatively detecting target substance to be detected in blood serum by utilizing light initiated chemiluminescence immune assay

A chemiluminescent immunoassay and quantitative detection technology, applied in the field of biology, can solve problems such as narrow quantitative range and limited quantitative detection range of immunological detection methods

Inactive Publication Date: 2013-02-27
李方和
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In addition, the quantitative detection range of immunological detection methods is usually relatively limited, and the quantitative range of the test method with higher sensitivity is usually narrower. Although luminescence analysis can correct this phenomenon to a certain extent, however, taking into account the sensitivity and specificity of the method. On the premise of stability, stability and reagent cost, its quantitative detection range is only about 1000ng / ml

Method used

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  • Method for qualitatively and quantitatively detecting target substance to be detected in blood serum by utilizing light initiated chemiluminescence immune assay
  • Method for qualitatively and quantitatively detecting target substance to be detected in blood serum by utilizing light initiated chemiluminescence immune assay
  • Method for qualitatively and quantitatively detecting target substance to be detected in blood serum by utilizing light initiated chemiluminescence immune assay

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1 The method for detecting HBsAg

[0045] 1. Detection method:

[0046] Method name: HBsAg detection target antigen superposition LiCA;

[0047] Original method name: HBsAg detection LiCA;

[0048] Sensitivity: 0.05-1.0ng / ml;

[0049] Quantitative range: within 0.1-500 to 1000ng / ml;

[0050] The starting point of the Hooks effect (that is, the starting point of the concentration-dependent reduction of the reaction signal): about 50 000 ng / ml;

[0051] False negative concentration (reaction signal close to cut off value): greater than 1 000 μg / ml;

[0052] 2. Materials:

[0053] 1. Instruments and consumables

[0054] Detection instrument: LiCA HT light-excited chemiluminescence detector; manufactured by China Boyang Bio (Shanghai) Technology Co., Ltd.;

[0055] 2. Reagents

[0056] (1) Photoactivated chemiluminescence kit for detection of serum HBsAg: manufactured by Boyang Bio (Shanghai) Technology Co., Ltd., commercially available. Contains the fol...

Embodiment 2

[0142] Example 2 The method for detecting serum anti-HBc

[0143] 1. Detection method:

[0144] Method name: Anti-HBc detection target antigen superposition LiCA;

[0145] Original method name: anti-HBc detection LiCA;

[0146] Sensitivity: 0.10 IU / ml;

[0147] Quantitative range: within 250 IU / ml;

[0148] The starting point of the Hooks effect (that is, the starting point of the concentration-dependent reduction of the reaction signal): about 2 500 IU / ml;

[0149]False negative concentration (reaction signal close to cut off value): greater than 200 000 IU / ml;

[0150] 2. Instruments and materials:

[0151] 1. Instruments and consumables

[0152] With embodiment 1.

[0153] 2 reagents

[0154] (1) Serum anti-HBc detection light-excited chemiluminescence kit: manufactured by Boyang Bio (Shanghai) Technology Co., Ltd., commercially available, including the following reagents:

[0155] Reagent 1: HBcAg-coated luminescent microspheres, the concentration is 10 μg / ml;

...

Embodiment 3

[0199] Example 3 The method for detecting serum AFP

[0200] one. Detection method:

[0201] Patented method name: Serum AFP detection target antigen superposition LiCA;

[0202] Original method name: serum AFP detection LiCA;

[0203] Sensitivity: 1.0 ng / ml;

[0204] Quantitative range: within 1000ng / ml;

[0205] The starting point of Hooks effect (that is, the starting point of the concentration-dependent reduction of the reaction signal): about 50 000 ng / ml;

[0206] False negative concentration (reaction signal close to cut off value): greater than 1 000 000ng / ml;

[0207] Instruments and materials:

[0208] 1. Reagents

[0209] (1) Serum AFP detection light-excited chemiluminescence kit: manufactured by Boyang Bio (Shanghai) Technology Co., Ltd., commercially available. Contains the following reagents:

[0210] Reagent 1: anti-AFP coated luminescent microspheres, the concentration is 100 μg / ml;

[0211] Reagent 2: biotin-labeled anti-AFP, the concentration is 1...

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Abstract

The invention discloses a method for qualitatively and quantitatively detecting a target substance to be detected in blood serum by utilizing light initiated chemiluminescence immune assay, and the method comprises the following steps of placing luminous microspheres coating a substance antibody, biotin-coated target substance antibody and optical sensitization microspheres wrapped by streptavidin (SA) into a specimen to be detected to perform a primary immunity reaction and detection, and the method also comprises the step of placing anti-He agent into the reaction system to facilitate the target substance immune superimposition reaction, and carrying out the secondary optical excitation chemicalluminescence immune detection. A purpose for comprehensively correcting a hooks effect can be realized by comprehensively analyzing detection results of twice light initiated chemiluminescence assay (LiCA), and the analysis process comprises the following steps of classifying the specimen to be detected into five concentration intervals such as cathode, low anode, middle anode, high anode and ultrahigh anode according to the signal characteristics of the primary LiCA detection and the secondary LiCA detection; and quantitatively analyzing the specimen in the low anode concentration interval and the ultrahigh anode concentration interval according to the primary LiCA detection. The method has the characteristics of accuracy in result, simplicity and convenience in operation, wide application range and the like.

Description

technical field [0001] The invention relates to a labeling immunoassay method in the field of biology, in particular to a method for qualitatively and quantitatively detecting target substances in serum by using light initiated chemiluminescence (Light initiatd Chemiluminescence Assay, LiCA) immunoassay. Background technique [0002] Homogeneous immunoassay refers to an immunological detection method in which all reactions and results are observed at a uniform liquid phase interface. Simple operation is the main feature of homogeneous immunoassay. Light-induced chemiluminescence (LiCA) immunoassay is the main representative of this type of method, and it is also one of the main homogeneous immunoassays that have been commercialized. Compared with analysis, LiCA immunoassay involves time-resolved fluorescence, nanosphere research, receptor (biotin and avidin) immobilization, special energy transfer mode and homogeneous immune response and other cutting-edge scientific achieve...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N21/76
Inventor 李方和李时君
Owner 李方和
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