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Chromogranin A chemiluminescence immunoassay reagent kit and preparation method thereof

A chemiluminescent immunoassay and chromogranin technology, which is applied in the field of immunoassay medicine, can solve problems such as radioactive pollution and inconsistent with the development trend of biomedical technology

Active Publication Date: 2014-07-23
天津市协和医药科技集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the radioactive pollution of iodine-125, radioimmunoassay technology is obviously not in line with the development trend of biomedical technology in today's increasingly demanding environment.

Method used

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  • Chromogranin A chemiluminescence immunoassay reagent kit and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0068] A preparation method and experimental operation procedure of a chromogranin A chemiluminescent immunoassay kit:

[0069] 1. Preparation of horseradish peroxidase-labeled chromogranin A monoclonal antibody (enzyme-labeled antibody for short):

[0070] ①Weigh 2.0mg of CgA monoclonal antibody into a 5ml glass bottle, add 1.0ml of 0.05mol / L sodium carbonate buffer solution with pH=7.5 to dissolve, and store at 4°C for later use;

[0071] ②Weigh 10.0mg of horseradish peroxidase into a 16×100mm glass test tube, add 1.0ml of deionized water, shake, and after it is completely dissolved, take out 0.14ml and put it into another 16×100mm glass test tube In the test tube, add 0.1mol / L NaIO 4 Solution 0.2ml (NaIO 4 The solution must be freshly prepared), the mouth of the glass test tube was tightly plugged with a rubber stopper, and the reaction was carried out at room temperature in the dark for 1 hour;

[0072] ③ Mix the liquid obtained in step ① with the liquid obtained in ste...

Embodiment 2

[0114] A preparation method and experimental operation procedure of a chromogranin A chemiluminescent immunoassay kit:

[0115] 1. Preparation of horseradish peroxidase-labeled chromogranin A monoclonal antibody (enzyme-labeled antibody for short):

[0116] ①Weigh 2.0mg of CgA monoclonal antibody into a 5ml glass bottle, add 1.5ml of 0.05mol / L sodium carbonate buffer solution with pH=7.5 to dissolve, and store at 4°C for later use;

[0117] ②Weigh 10.0mg of horseradish peroxidase into a 16×100mm glass test tube, add 1.0ml of deionized water, shake, after it is completely dissolved, take out 0.21ml and put it into another 16×100mm glass test tube In the test tube, add 0.1mol / L NaIO 4 Solution 0.2ml (NaIO 4 The solution must be freshly prepared), the mouth of the glass test tube was tightly plugged with a rubber stopper, and the reaction was carried out at room temperature in the dark for 1.5 hours;

[0118] ③ Mix the liquid obtained in step ① with the liquid obtained in step...

Embodiment 3

[0153] A preparation method and experimental operation procedure of a chromogranin A chemiluminescent immunoassay kit:

[0154] 1. Preparation of horseradish peroxidase-labeled chromogranin A monoclonal antibody (enzyme-labeled antibody for short):

[0155] ①Weigh 2.0mg of CgA monoclonal antibody into a 5ml glass bottle, add 2.0ml of 0.05mol / L sodium carbonate buffer solution with pH=7.5 to dissolve, and store at 4°C for later use;

[0156] ② Weigh 10.0mg of horseradish peroxidase into a 16×100mm glass test tube, add 1.0ml of deionized water, shake, and after it is completely dissolved, take out 0.28ml and put it into another 16×100mm glass test tube In the test tube, add 0.1mol / L NaIO 4 Solution 0.2ml (NaIO 4 The solution must be freshly prepared), the mouth of the glass test tube was tightly plugged with a rubber stopper, and the reaction was carried out at room temperature in the dark for 2 hours;

[0157] ③ Mix the liquid obtained in step ① with the liquid obtained in s...

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Abstract

The invention discloses a chromogranin A chemiluminescence immunoassay reagent kit and a preparation method of the reagent kit. The reagent kit comprises the following ingredients: 1) chromogranin A standard products; 2) horseradish peroxidase labeled chromogranin A monoclonal antibodies; 3) chromogranin A monoclonal antibody covered magnetic particles; 4) white non-transparent micro porous plate; 5) cleaning solution and 6) chemiluminescence substrate A and B work solution. The chemiluminescence immunoassay and the nanometer magnetic particle technology are combined, and the chromogranin A chemiluminescence immunoassay reagent kit is provided. The defects of low sensitivity and poor accuracy of the traditional enzyme-linked immunosorbent assay (ELISA) method are overcome, the radioactive pollution of the radioimmunoassay (RIA) technology is avoided, and the chromogranin A chemiluminescence immunoassay reagent kit and the preparation method thereof have the advantages that safety is realized, the environment is protected, the specificity is high, the sensitivity and the accuracy are high, and the like.

Description

technical field [0001] The invention relates to the technical field of immunoanalysis medicine, in particular to a chemiluminescent immunoassay kit for chromogranin A (CgA) in human serum and a preparation method thereof. Background technique [0002] Chromogranin A (CgA) is an acidic soluble protein composed of 439 amino acids, and is the main member of the chromogranin family. Extensively present in neuroendocrine cells, elevated concentrations of CgA in plasma may indicate tumors of neuroendocrine origin. Elevated CgA levels are seen in the diagnosis of almost all types of neuroendocrine tumors. Therefore, in the diagnosis of neuroendocrine tumors, CgA is a very valuable diagnostic marker. If 32.7U / L was used as the cut-off value, the sensitivity and specificity of CgA to pancreatic tumors were 82.1% and 96.2%, respectively, and the sensitivity and specificity to pheochromocytoma were 88.5% and 96.2%, respectively. Therefore, the quantitative detection of CgA is of gre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/531G01N21/76
Inventor 王立凯潘学继单存海
Owner 天津市协和医药科技集团有限公司
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