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Liver cancer tumour marker AFP, AFP variant and GP73 antigen synchronous immunodetection kit, method and application

A tumor marker, alpha-fetoprotein technology, applied in biological tests, measurement devices, material inspection products, etc., can solve the problems of missed detection and false positives, and achieve the effect of wide clinical application value

Active Publication Date: 2019-10-11
万东山
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] As a newly discovered diagnostic marker for liver cancer, GP73 has higher diagnostic sensitivity and specificity than AFP. However, there are certain false positives and missed detections in a single inspection. The combined inspection of multiple tumor markers is effective for Improving the sensitivity and specificity of liver cancer diagnosis is of great significance

Method used

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  • Liver cancer tumour marker AFP, AFP variant and GP73 antigen synchronous immunodetection kit, method and application
  • Liver cancer tumour marker AFP, AFP variant and GP73 antigen synchronous immunodetection kit, method and application
  • Liver cancer tumour marker AFP, AFP variant and GP73 antigen synchronous immunodetection kit, method and application

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preparation example Construction

[0062] Preferably, in the kit according to the first aspect of the present invention, the magnetic particles are magnetic particles containing toluenesulfonyl groups with a particle size of 2.8 μm. The inventors have found that such magnetic particles have a high labeling rate on the magnetic particles of the present invention, gather rapidly after applying magnetic force, and have good adsorption consistency. In a particular embodiment of the invention, the magnetic particles are Dynabeads M-270 type magnetic particles. More preferably, in the kit according to the first aspect of the present invention, the preparation method of antibody-labeled magnetic particles includes: adding antibodies to magnetic particles, vibrating labeling, adding BSA to block, discarding the supernatant, and then washing.

[0063] Preferably in the kit of the invention the acridinium ester is acridinium succinimide ester. The inventors of the present invention have found that the acridinium ester b...

Embodiment 1

[0076] Example 1 Preparation of detection kits for alpha-fetoprotein (AFP), alpha-fetoprotein variant (AFP-L3), and Golgi protein 73 (GP73)

[0077] After research by the inventors, AFP monoclonal antibody (available from sigma, product number SAB420074) was selected to label magnetic particles, and AFP monoclonal antibody antibody (available from sigma, product number GW22680) was selected to label acridinium ester.

[0078] After research by the inventors, LCA (available from sigma, product number L1277) was selected to label the magnetic particles, and the fifth fragment of bovine serum albumin (available from sigma, product number B064) was selected to label LCA.

[0079] After research by the inventors, the magnetic particles were labeled with GP73 monoclonal antibody (available from Santa Cruz, product number Goy-kt0635), and acridinium ester was labeled with GP73 polyclonal antibody (available from Abcam, product number yb-668Hu01).

[0080] Although it is not the antib...

Embodiment 2

[0124] Example 2 Determination of AFP, AFP-L3, and GP73 kits

[0125] 1. Determination of magnetic particles: Dilute AFP, LCA-BSA, and GP73 magnetic particles at appropriate concentrations.

[0126] 2. Determination of acridinium esters: Mix AFP and GP73-labeled acridinium esters at a ratio of 1:1, and dilute to an appropriate concentration with diluent.

[0127] 3. The simultaneous detection kit of the present invention can include 13 containers, respectively equipped with prepared AFP, LCA-BSA, GP73 labeled magnetic particles, prepared acridinium ester conjugates and AFP, GP73 calibrator S0 ~ S5, In addition, the detection kit of the present invention may also include containers respectively containing the following reagents: washing solution (its formula is: 0.01M phosphate buffer solution pH7.4, 0.05% (v / v) Tween 20), excitation solution A (its formula is: containing 0.05M nitric acid solution, 0.15% (v / v) H 2 o 2 ) and excitation solution B (the formula is: 0.2M NaOH s...

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Abstract

The invention relates to a liver cancer tumour marker AFP, AFP variant and GP73 antigen synchronous immunodetection kit, a method and application, and belongs to the technical field of immunodetection. The kit comprises magnetic particles, AFP and GP73 antibody coupled mixed acridinium ester, GP73, an AFP calibration product, first stimulation solution, second stimulation solution, washing solution and all kinds of diluting solution; and the magnetic particles are respectively marked by AFP, LCA-BSA and GP73. By adoption of the kit and the detection method, the sensitivity, the specificity andthe accuracy are high; the detection time is short; identification on benign liver diseases and malignant liver diseases and early diagnosis on liver cancers can be easily carried out; and the kit and the detection method have an important purpose for increasing the early diagnosis rate of patients with liver cancers and prolonging the survival rate, and have wide clinical application value.

Description

technical field [0001] The invention belongs to the technical field of immune detection, and in particular relates to a simultaneous immunological detection kit, method and application of liver cancer tumor marker AFP, alpha-fetoprotein heterogeneity and GP73 antigen. Background technique [0002] Liver cancer is one of the most common malignant tumors in my country, with a five-year survival rate of less than 10%, and has become the second leading cause of death from malignant tumors in my country. The early symptoms of liver cancer are not obvious. Once discovered, it is often late and loses the best clinical treatment period. At present, the clinical diagnosis of liver cancer mainly adopts the method of combining imaging examination and serum marker examination. The sensitivity of serum marker examination It is higher than imaging, and is simple to operate and low in price. It is suitable for large-scale screening of high-risk groups. Alpha-fetoprotein, alpha-fetoprotein ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/58G01N33/574G01N33/543
CPCG01N33/6893G01N33/57484G01N33/57438G01N33/585G01N33/54326
Inventor 万东山
Owner 万东山
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