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Liver cancer multi-marker micro-array kit as well as preparation method and application thereof

A multi-marker, microarray technology, applied in the biological field, can solve problems such as difficult to distinguish benign from malignant lesions

Active Publication Date: 2011-06-01
THE FIFTH MEDICAL CENT OF CHINESE PLA GENERAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

AFP is often elevated in patients with primary liver cancer, but many benign liver diseases may also have elevated AFP, sometimes it is difficult to distinguish benign and malignant lesions based on AFP results alone

Method used

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  • Liver cancer multi-marker micro-array kit as well as preparation method and application thereof
  • Liver cancer multi-marker micro-array kit as well as preparation method and application thereof
  • Liver cancer multi-marker micro-array kit as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Quality detection of the chemiluminescent quantitative enzyme-linked immunoassay kit of the present invention

[0046] 1) Accuracy: In the test results of 15 negative quality control sera (including specific control sera) in normal physical examination, no false positives occurred. There were no false negatives in the test results of 10 reference samples of liver cancer HCC positive quality control serum.

[0047] 2) Precision: 20 boxes of different batches of kits were randomly selected, and the same HCC antigen-positive quality control serum was used for repeated measurements according to the instructions. Calculate the results of each measurement, find the mean, SD and coefficient of variation CV. The results of the precision test showed that the CV between batches was <15%.

[0048] 3) Detection sensitivity: the detection sensitivity of this kit is 5ng / ml.

[0049] 4) Specificity: Take four samples to be tested and mix them into four mixed serum sample...

Embodiment 2

[0051] Example 2: Preparation of liver cancer multi-marker microarray chemiluminescence kit

[0052] This embodiment prepared a liver cancer multi-marker microarray chemiluminescence kit (96 servings) of the present invention, which consists of:

[0053] AFP / GP73 monoclonal antibody coated plate (96 wells) 1 piece;

[0054] 1 bottle of horseradish peroxidase (HRP)-labeled AFP antibody, 1 bottle of horseradish peroxidase (HRP)-labeled GP73 polyclonal antibody, 1 bottle of horseradish peroxidase (HRP)-labeled lectin LCA solution , respectively 10ml / bottle;

[0055] 1 bottle of substrate solution: EDTA (1.0×10 -2 M), H 2 o 2 (7.5×10 -3 M), HCl (1.0×10 -2 M) and Tween20 (1%)

[0056] Chromogenic solution 1 bottle: luminol 5.0×10 -4 Mol / L;

[0057] 1 bottle of sample diluent: 20mM pH 7.4 PBS buffer containing 2% BSA

[0058] The specific operation is as follows:

[0059] 1. Make AFP / GP73 monoclonal antibody coated plate:

[0060] The enzyme plate adopts the 12×8 detacha...

Embodiment 3

[0066] Example 3: The method of using the multi-marker microarray chemiluminescent immunoassay kit for liver cancer

[0067] a) Monoclonal antibody capture antigen reaction: add 100 μl sample diluent to the corresponding microwells of the antibody-coated plate provided in the kit, then add 10 μl serum sample to be tested, incubate in a 37°C water bath for 30 minutes, and wash the plate. A total of 4 well reactions, respectively Al, Cl, El, Gl;

[0068]d) Add AFP-mAb-HRP solution to Al, add LCA-HRP solution to Cl well, add GP73 mAb-HRP solution to El well, 100 μl per well, and keep warm in 37° C. water bath for 30 minutes. Repeat the plate washing operation 4 times.

[0069] c) Chromogenic reaction: add substrate solution A and 50 μl of chromogenic solution B to each well successively, and read the value.

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Abstract

The invention provides a protein micro-array kit for early diagnosis of liver caner as well as a preparation method and an application thereof. The kit can be used for monitoring of development level of chronic hepatitis and prognosis, as well as early diagnosis of the liver cancer. The kit can be used for monitoring expression levels of alpha-fetoprotein, alpha-fetoprotein variant, Golgi protein GP73 and Golgi protein GP73 variants, and determining the prognosis level of a patient and the probability of causing cirrhosis and the liver cancer, thereby providing a direct support for the prevention, diagnosis and treatment of the liver cancer and the hepatitis.

Description

technical field [0001] The invention belongs to the field of biotechnology. More specifically, the invention provides a protein microarray detection kit for alpha-fetoprotein and its heterogeneity, Golgi protein GP73 and its heterogeneity, its preparation method and application. The kit can be used to quickly and accurately calculate the expression levels of alpha-fetoprotein and its variants, Golgi protein GP73 and its variants in human peripheral blood. Background technique [0002] The main etiology of HCC (hepatocellular carcinoma) is HBV or HCV chronic liver virus infection, and there is generally a long incubation period before the occurrence of liver cancer—that is, the time from infection of liver virus to occurrence of liver cancer. According to the report of "China Medical News" on January 26, 2006 and the introduction of the article in the sixth issue of "Cancer" in 2005, there are about 120 million hepatitis B virus carriers in my country, accounting for about 1 / ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/574
Inventor 李伯安毛远丽侯俊徐军孙志强郭桐生
Owner THE FIFTH MEDICAL CENT OF CHINESE PLA GENERAL HOSPITAL
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