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Chemiluminescent quantitative detection kit for blood plasma HSP70 (heat shock protein) antibody

A chemiluminescence and quantitative detection technology, which is applied in the field of in vitro clinical testing and chemiluminescence immunoassay, can solve the problems of detection sensitivity, professionalism, equipment and facility conditions and cost impact, and achieve easy standardized operation, improved sensitivity and high sensitivity Effect

Inactive Publication Date: 2014-09-17
GUANGXI DOCTOR HAIYI INFORMATION TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Various methods confirm each other, and at the same time, there are influences on detection sensitivity, professionalism, equipment and facility conditions and cost

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1: Preparation of the plasma HSP70 antibody chemiluminescence quantitative detection kit of the present invention

[0017] 1. Preparation of Solid-Phase Coated Plates

[0018] 1) Coating: use 0.05mol / L sodium carbonate buffer solution with pH 9.6, add an appropriate amount of recombinant HSP70 antigen and mix well, then add to the wells of a microplate, 100 μl / well, overnight at 4°C;

[0019] 2) Sealing: discard the coating solution, pat dry on absorbent paper, add 3% BSA and 0.05% preservative (Proclin TM 300) of phosphate buffer (pH 7.4), 200 μl / well, 2 hours at 37°C;

[0020] 3) Seal the bag: Discard the sealing solution, pat it dry on absorbent paper, pump it in a vacuum oven for 5 hours at room temperature, immediately vacuum seal the bag, check for air leaks, and re-seal the bag if necessary, after labeling Store at 2-8°C.

[0021] 2. Preparation of enzyme-labeled anti-HSP70 antibody antibody (enzyme-labeled secondary antibody)

[0022] The antibody a...

Embodiment 2

[0033] Embodiment 2: the using method of kit of the present invention

[0034] The specific operation of the plasma HSP70 antibody chemiluminescence quantitative detection kit prepared in Example 1 is as follows:

[0035] 1. Reagent and sample preparation

[0036] 1) Reagent preparation

[0037] ①The kit should be placed at room temperature (18-26°C) to equilibrate for 20 minutes.

[0038] ② Take out the concentrated washing liquid from the kit, dilute it with purified water 1:20, and add it to the washing liquid bottle of the plate washer.

[0039] 2) Sample preparation

[0040] Place the qualified serum or plasma to be tested at room temperature (18-26°C) to equilibrate for 20 minutes before use.

[0041] 2. Test conditions

[0042] Operate at room temperature (18-26°C) strictly according to the instructions of the kit.

[0043] 3. Operation steps

[0044] 1) Take out the required recombinant HSP70 antigen-coated microwell plate reaction strips and place them on the m...

Embodiment 3

[0059] Embodiment 3: the methodological identification of the kit of the present invention

[0060] Test the kit prepared in Example 1 according to the conventional manufacturing and testing process in the art:

[0061] 1. Sensitivity detection of plasma HSP70 antibody chemiluminescence quantitative detection kit

[0062] Using the plasma HSP70 antibody chemiluminescence quantitative detection kit prepared in Example 1, different concentrations of plasma HSP70 antibody antigens were added to the microwells of the microwell plate, and the concentrations of plasma HSP70 antibody antigens were 0, 0.25, 1, 5, and 10 respectively. , 25, 50 ng / ml.

[0063] After conversion, the sensitivity of this kit reaches 0.1ng / ml

[0064] 2. Specific detection of plasma HSP70 antibody chemiluminescence quantitative detection kit

[0065] Using the plasma HSP70 antibody chemiluminescence quantitative detection kit prepared in Example 1, add other antibodies in plasma to the microwells of the ...

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Abstract

The invention discloses a chemiluminescent quantitative detection kit for a blood plasma HSP70 (heat shock protein) antibody, belonging to the field of external clinical inspection and chemiluminescent immunodetection. The kit comprises a blood plasma HSP70 antibody detection reaction plate which is coated with a recombined HSP70 antigen; the quantitative detection kit for the plasma HSP70 antibody further comprises an enzyme conjugate, a luminous substrate, a calibration material, a quality control substance and concentrated washing liquid. The kit disclosed by the invention can exclusively and quantitatively detect the content of the blood plasma HSP70 antibody; compared with the conventional ELISA (enzyme-linked immuno sorbent assay) technology, the chemiluminescent quantitative detection kit has the advantages that the high specificity of the ELISA technology, and the stability, the reliability and the operation convenience of a detection result are retained, and meanwhile, by the adoption of a chemiluminescent method, the detection sensitivity is improved.

Description

technical field [0001] The invention belongs to the fields of in vitro clinical testing and chemiluminescence immunoassay, and in particular relates to a kit for quantitatively detecting plasma HSP70 antibody. Background technique [0002] The heat shock protein family (HSPs) is a class of highly conserved proteins, and HSP70 is an important member of the heat shock protein family, which is divided into inducible and constitutive. When the body is stimulated by various adverse factors in the external environment, the expression of inducible HSP70 protein increases. HSP70 proteins are essential for the repair of cell damage, survival and maintenance of normal cell function. It acts as a molecular chaperone in environmental stimuli, pathogen infection, and cell stress damage caused by diseases, preventing protein aggregation and promoting the refolding of damaged proteins. In most mammals, inducible HSP70 is expressed only under stress conditions and can be detected after si...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/535
CPCG01N33/96G01N21/76G01N33/535
Inventor 李永锋陈国勇傅汝毅陶玲云蓝文苑
Owner GUANGXI DOCTOR HAIYI INFORMATION TECH
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