Enzyme-linked immunosorbent assay kit for detecting concentration of tumor marker DKK1

Abstract

The invention discloses an enzyme-linked immunosorbent assay kit for detecting the concentration of the tumor marker DKK1. The enzyme-linked immunosorbent assay kit is composed of a porous plate coated with DKK1 monoclonal antibodies 1 with the model of 10C3, monoclonal antibodies 2 with the enzyme labeling model of 6E8, a chromogenic substrate and a substrate stop solution. The invention further discloses a preparation method and performance indexes of the enzyme-linked immunosorbent assay kit. The enzyme-linked immunosorbent assay kit matched with existing instruments in most hospitals is prepared by adopting the totally-domestic reagents, ultrahigh-sensitivity, high-specificity and low-cost detection of the concentration of DKK1 is achieved, large-scale general investigation of hepatocellular carcinoma is facilitated, and diagnosis can be carried out as early as possible to improve the health of all people of China.

Description

technical field [0001] The invention relates to the technical field of tumor diagnosis, in particular to an enzyme-linked immunoassay kit for detecting the concentration of a tumor marker DKK1. Background technique [0002] Worldwide, hepatocellular carcinoma (HCC) is the sixth most common malignancy and the third leading cause of cancer-related death. In my country, the high-risk population of hepatocellular carcinoma, including long-term carriers of various hepatitis viruses and patients with liver cirrhosis, is as high as 150 million. The 5-year overall survival rate of HCC is only 3-5%. Part of the reason for this extremely low survival rate is the lack of a timely and effective diagnosis, resulting in 60-70% of HCC being diagnosed at the time of diagnosis. Patients missed the best opportunity for treatment. [0003] The serum protein DKK1 discovered by the Shanghai Cancer Institute is a better marker than the existing hepatocellular carcinoma diagnostic marker alpha-f...

AI Technical Summary

Problems solved by technology

Chinese patent CN101398433 discloses a kit based on a time-resolved fluorescent immunoassay method, which requires a time-resolved fluorescent immunoassay analyzer that is not equipped in most hospitals, and this method also requires the use of relatively expensive fluorescent markers , is not conducive to a large-scale census of DKK1

Chinese patent CN102507951 discloses an enzyme-linked immunoassay kit for joint detection of tumor markers, which is coated with neuron-specific enolase (NSE), cytokeratin fragment 21-1 (CYFRA21-1) and Dkkopf1 (DKK1) multi-well plate of monoclonal antibody, horseradish peroxidase-labeled NSE, CYFRA21-1 and DKK1, chromogenic substrate TMB and substrate stop solution; each well of the multi-well plate is coated with NSE monoclonal antibody 1 -4ug / ml; each well is coated with CYFRA21-1 monoclonal antibody 1-2ug / ml; each well is coated with DKK1 monoclonal antibody 1-2ug / ml; NSE, CYFRA21-1, DKK1 three kinds of detection antibodies according to 4: The ratio of 4:2 is added to the wells of the multi-well plate, and the total volume is 100ul; although the kit disclosed by the invention can detect two or three tumor-associated antigens at the same time, due to the dispersion of the detection accuracy and sensitivity of each antigen, and for The detection of DKK1 antigen only uses a DKK1 antibody, so the detection accuracy of DKK1 antigen is difficult to meet the rapid and accurate detection of tumors

In addition, although there are enzyme-linked immunosorbent assay kits for detecting DKK1 protein in foreign countries (Cat. No. DKK100, R&D Systems, the United States), they are expensive (~￥6000 / kit), which is not conducive to the detection of DKK1 by many high-risk groups of hepatocellular carcinoma in my country. Size Census Needs

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