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Squamous cell carcinoma antigen chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof

A technique for chemiluminescence immunity and squamous cell carcinoma, which is applied in the field of immunoassay medicine and can solve the problems of not meeting clinical requirements, false negatives, false positives, etc.

Inactive Publication Date: 2009-03-04
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] For the quantitative analysis of SCC, the currently commonly used method is enzyme-linked immunosorbent assay (ELISA), but because the detection sensitivity of enzyme-linked immunosorbent assay (ELISA) is not high enough, the detection range is narrow, and there are many influencing factors, it is easy to cause false negatives. and false positives, so it cannot meet the clinical requirements

Method used

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  • Squamous cell carcinoma antigen chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof
  • Squamous cell carcinoma antigen chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof
  • Squamous cell carcinoma antigen chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof

Examples

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Embodiment 1

[0028] Example 1 Preparation of squamous cell carcinoma antigen chemiluminescence immunoassay quantitative assay kit of the present invention using alkaline phosphatase system

[0029] Firstly, the screening test and quality identification of the raw materials used were carried out to ensure the accuracy of the test, and then the coating method was studied, the most suitable coating buffer and blocking solution were selected, and the optimal concentration conditions were found, and Enzyme marker dilutions that maintain activity of the enzyme marker over a long period of time are preferentially formulated.

[0030] 1. Preparation of alkaline phosphatase-labeled antibody

[0031] The classic glutaraldehyde cross-linking method was used to couple with alkaline phosphatase, fully dialyzed against PBS, added an equal volume of glycerol, and stored in a low-temperature refrigerator (-20°C). Dilute with Enzyme Label Diluent before use. It has been proved by experiments that the wor...

Embodiment 2

[0077] Example 2 Application of horseradish peroxidase system to prepare squamous cell carcinoma antigen chemiluminescent immunoassay quantitative assay kit of the present invention

[0078] 1. Preparation of horseradish peroxidase-labeled antibody

[0079] Dissolve 4.4mg HRP in 1mL distilled water, add 0.4mL sodium periodate (50mmol / L) and stir at room temperature for 20min, dialyze through 1mmol / L sodium acetate buffer solution, pH value is 4.4, add 8mg squamous cell carcinoma antigen antibody, stir 2h, finally use 200mmol / L NaBH 4 For reduction, after dialysis with 0.02M PBS buffer, add an equal volume of glycerol, and store below -20°C.

[0080] 2. Preparation of chemiluminescent substrates

[0081] Chemiluminescence Substrate A Solution

[0082] Luminol 10mM 1.7716g

[0083] 4-Hydroxybiphenyl 0.3mM 0.051g

[0084] 4-iodophenylboronic acid 0.05mM 0.012g

[0085] Boric acid 11.4g

[0086] Borax 4.9g

[0087] Double distilled water fixed solution 1000mL

[0088] Adj...

Embodiment 2～3

[0106] Examples 2-3 Preparation of squamous cell carcinoma antigen chemiluminescent immunoassay quantitative assay kit of the present invention

[0107] The quantitative assay kit of the present invention was prepared in the same manner as in Example 1 except that plastic tubes and plastic beads were used as carriers respectively.

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Abstract

The invention provides a chemiluminescent immunoassay quantitative detection kit for squamous cell carcinoma antigens, which relates to the medical field of immunoassay. The kit of the invention is mainly composed of a squamous cell carcinoma antigenic temporary calibrator, a solid-phase vector which is coated by a squamous cell carcinoma antigenic antibody, a squamous cell carcinoma antigenic antibody which is marked by enzyme, a chemiluminescent substrate solution and a concentrated washing solution. The preparation method of the kit mainly comprises the following steps: the coating of the solid-phase vector, the preparation of the calibrator and enzyme markers and the preparation of the chemiluminescent substrate solution and the concentrated washing solution. With the kit and the preparation method thereof, the detection sensitivity, the linear range and the quantitative accuracy can be improved. The invention has convenient use, safety and stability.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine, in particular, the invention provides a squamous cell carcinoma antigen chemiluminescent immunoassay quantitative assay kit and a preparation method thereof. Background technique [0002] Squamous cell carcinoma antigen (SCC) is a tumor marker with good specificity and the earliest diagnosis of squamous cell carcinoma. It was originally isolated from cervical cancer tissue and belongs to the family of serine protease inhibitors in terms of biological activity, which includes two genes, SCC1 and SCC2. SCC widely exists in normal tissues of different organs (minimal content) and in epithelial cells of malignant lesions. There are at least four forms of SCC in serum: free SCC1, free SCC2, and the corresponding serine protease conjugates. SCC inhibits cell apoptosis and participates in the differentiation of squamous epithelial layer in normal squamous epithelial cells, and participates in tu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/543G01N21/76
Inventor 任超应希堂宋胜利胡国茂郑金来唐宝军
Owner CHEMCLIN DIAGNOSTICS CO LTD
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