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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 chemiluminescence immunoassay and quantitative detection technology, applied in the field of biology, can solve the problems of narrow quantitative range and limited quantitative detection range of immunological detection methods.

Inactive Publication Date: 2015-05-20
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

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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

[0045] Example 1 Method for detecting HBsAg

[0046] 1. Detection method:

[0047] Method name: HBsAg detection target antigen plus LiCA;

[0048] Original method name: HBsAg detection LiCA;

[0049] Sensitivity: 0.05-1.0ng / ml;

[0050] Quantitative range: 0.1-500 to 1000ng / ml;

[0051] Hooks effect starting point (that is, the starting point for the reaction signal to decrease in a concentration-dependent manner): about 50 000 ng / ml;

[0052] Concentration of false negative appearance (reaction signal is close to cut off value): greater than 1 000μg / ml;

[0053] 2. Material:

[0054] 1. Instruments and consumables

[0055] Detection instrument: LiCA HT light-induced chemiluminescence detector; manufactured by China Boyang Biological (Shanghai) Technology Co., Ltd.;

[0056] 2. Reagent

[0057] (1) Serum HBsAg detection light-induced chemiluminescence kit: manufactured by Boyang Biological (Shanghai) Technology Co., Ltd. and commercially available. Includes the following reagents:

[0058] Rea...

Embodiment 2

[0144] Example 2 Method of detecting serum anti-HBc

[0145] 1. Detection method:

[0146] Method name: Anti-HBc detection target antigen plus LiCA;

[0147] Original method name: LiCA for anti-HBc detection;

[0148] Sensitivity: 0.10IU / ml;

[0149] Quantitative range: within 250IU / ml;

[0150] The starting point of the Hooks effect (that is, the starting point of a concentration-dependent decrease of the response signal): about 2 500IU / ml;

[0151] Concentration of false negative appearance (reaction signal is close to cut off value): greater than 200 000IU / ml;

[0152] 2. Instruments and materials:

[0153] 1. Instruments and consumables

[0154] The same as in Example 1.

[0155] 2 reagent

[0156] (1) Serum anti-HBc detection light-excited chemiluminescence kit: manufactured by Boyang Biological (Shanghai) Technology Co., Ltd., commercially available, including the following reagents:

[0157] Reagent 1: HBcAg coated luminescent microspheres at a concentration of 10μg / ml;

[0158] Reagent 2...

Embodiment 3

[0201] Example 3 Method for detecting serum AFP

[0202] 1. Detection method:

[0203] Patent method name: Serum AFP detection target antigen superimposed LiCA;

[0204] Original method name: Serum AFP to detect LiCA;

[0205] Sensitivity: 1.0ng / ml;

[0206] Quantitative range: within 1000ng / ml;

[0207] Hooks effect starting point (that is, the starting point for the reaction signal to decrease in a concentration-dependent manner): about 50 000ng / ml

[0208] Concentration of false negative occurrence (reaction signal is close to cut off value): greater than 1 000 000 ng / ml;

[0209] Instruments and materials:

[0210] 1. Reagent

[0211] (1) Serum AFP detection light-excited chemiluminescence kit: manufactured by Boyang Biological (Shanghai) Technology Co., Ltd. and commercially available. Includes the following reagents:

[0212] Reagent 1: Anti-AFP coated luminescent microspheres at a concentration of 100μg / ml;

[0213] Reagent 2: Biotin-labeled anti-AFP at a concentration of 10.μg / ml;

[0...

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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 labeled immunoassay method in the biological field, and specifically relates to a method for qualitatively and quantitatively detecting target substances in serum by adopting light initiatd chemiluminescence Assay (LiCA) immunoassay. Background technique [0002] Homogeneous immunity refers to an immunological detection method in which all reactions and results are observed at a uniform liquid interface. Simple operation is the main feature of homogeneous immunity. 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. It is combined with enzyme-linked immunoassay (ELISA), electrochemiluminescence immunoassay and other solid-phase immunoassays. Compared with analysis, LiCA immunoassay involves time-resolved fluorescence, nano-microsphere research, receptor (biotin and avidin) immobilization, special...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53G01N21/76
Inventor 李方和李时君
Owner 李方和
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