Chromogranin A chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof

A chemiluminescence immunity and chemiluminescence technology, which is applied in the field of immunoanalysis medicine, can solve problems such as narrow detection range, false positives, and failure to meet clinical requirements, and achieve high sensitivity and stability, good specificity and accuracy

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

[0010] For the quantitative analysis of CgA, the commonly used methods are immunoradiometric assay (IRMA), radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA), although immunoradiometric assay and radioimmunoassay have high sensitivity , but it has disadvantages such as complex operation, unstable measurement results, short storage time of reagents, and radioactive contamination; while ELISA has insufficient detection sensitivity, narrow detection range, and many influencing factors, which can easily cause false negatives and false positives. Positive and other shortcomings
Therefore, none of the above methods can meet the clinical requirements.

Method used

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  • Chromogranin A chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof
  • Chromogranin A chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof
  • Chromogranin A chemiluminescence immune analysis quantitative determination reagent kit and preparing method thereof

Examples

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Effect test

Embodiment 1

[0027] Example 1 Preparation of Chromogranin A Chemiluminescent Immunoassay Quantitative Assay Kit of the Present Invention Using Alkaline Phosphatase System

[0028] 1. Preparation of alkaline phosphatase-labeled antibody

[0029] The CgA-specific antibody was conjugated with alkaline phosphatase by classical glutaraldehyde cross-linking method, fully dialyzed with PBS buffer, and an equal volume of glycerol was added to the conjugate solution and stored in a low-temperature refrigerator (-20°C). Dilute with 20% bovine serum enzyme diluent when used again. The optimal working concentration range of the enzyme-labeled antibody is 1:5000-10000 through optimization experiments.

[0030] The formula of the 20% bovine serum enzyme dilution used is:

[0031] NaH2PO4·2H2O 1.28mM 0.2g

[0032] Na2HPO4·12H2O 8.10mM 2.9g

[0033] NaCl 150mM 8.8g

[0034] Newborn Calf Serum 20% 200ml

[0035] Proclin300 0.1% 1.0ml

[0036] Food Red 0.05‰ 1.0ml

[0037] Dilute to 1000mL with deio...

Embodiment 2

[0074] Example 2 Application of horseradish peroxidase system to prepare chromogranin A chemiluminescent immunoassay quantitative assay kit of the present invention

[0075] 1. Preparation of horseradish peroxidase-labeled antibody

[0076] 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 chromogranin A specific antibody, Stir for 2h, and 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.

[0077] 2. Preparation of chemiluminescence substrate

[0078] 1. Chemiluminescent Substrate A Solution

[0079] Luminol 10mM 1.7716g

[0080] 4-Hydroxybiphenyl 0.3mM 0.051g

[0081] 4-iodophenylboronic acid 0.05mM 0.012g

[0082] Boric acid 11.4g

[0083] Borax 4.9g

[0084] Double distilled water fixed solution 1000mL

[0085] Adjust pH to 8.0~1...

Embodiment 3～4

[0103] Examples 3-4 Preparation of Chromogranin A Chemiluminescence Immunoassay Quantitative Assay Kit of the present invention

[0104] Except that plastic beads were used as the carrier, the rest were the same as those in Examples 1 and 2 for preparing the quantitative assay kit of the present invention.

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Abstract

The invention relates to a chromogranin A chemiluminescent quantitative detection kit, which relates to the medical field of immunoassay. The invention provides a chromogranin A chemiluminescent immunoassay quantitative detection kit and a preparation method thereof. The kit is mainly composed of a chromogranin A antigen calibrator, a solid-phase vector which is coated by chromogranin A antigen, a chromogranin A specific antibody which is marked by enzyme, chemiluminescent substrate solution and concentrated washing solution. The kit adopts the double antibody sandwich one step method to detect the content of chromogranin A in human serum, can effectively improve the sensitivity and the stability, and has good application value in clinic.

Description

technical field [0001] The invention mainly relates to the field of immunoanalysis medicine, and provides a chromaffin A chemiluminescent immunoassay quantitative assay kit and a preparation method thereof. Background technique [0002] Chromogranin A (CgA), also known as chromogranin A, chromogranin A, etc., is an acidic, hydrophilic protein composed of 439 amino acids, located in the chromaffin granules of neuroendocrine cells. It is a member of the neuropeptide family. Proteolysis of CgA is tissue-specific, and the fragmentation of this protein varies depending on the tissue in which it resides. The presence of CgA in tumor cells by immunohistochemistry is associated with neuroendocrine primary tumors, and serum CgA levels are elevated in patients with neuroendocrine tumors. Serum CgA is a better marker for neuroendocrine tumors today, therefore, the detection of serum CgA can be used for the diagnosis and treatment of neuroendocrine tumors. [0003] In terms of pheoch...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543G01N21/76
Inventor 任超应希堂胡国茂郑金来唐宝军于尚永
Owner CHEMCLIN DIAGNOSTICS CO LTD
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