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Protein chip reagent kit and method for comprehensively detecting lung cancer marker

A protein chip and comprehensive detection technology, which is applied in the field of protein chip kits for comprehensive detection of lung cancer markers, can solve the problem of insufficient specificity and sensitivity of lung cancer, and achieve the purpose of reducing experimental steps, saving time and cost, and improving accuracy. Effect

Inactive Publication Date: 2013-04-10
河南生生医疗器械有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Currently, markers for detecting lung cancer include SCC (squamous cell carcinoma-associated antigen), CEA (carcinoembryonic antigen), CA19-9 (carbohydrate antigen), CYFRA21-1 (cytokeratin 19 fragment), NSE (neuron-specific Enolase), pro-GRP (pro-gastrin-releasing peptide), the specificity and sensitivity of each marker in the diagnosis of lung cancer are not high enough, and the combination of these six indicators in the diagnosis of lung cancer can improve the specificity and sensitivity of diagnosis , but currently clinically used are products that detect a single indicator

Method used

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  • Protein chip reagent kit and method for comprehensively detecting lung cancer marker
  • Protein chip reagent kit and method for comprehensively detecting lung cancer marker
  • Protein chip reagent kit and method for comprehensively detecting lung cancer marker

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Embodiment 1 Preparation of a protein chip kit for comprehensive detection of lung cancer markers of the present invention:

[0026] 1. Chip production:

[0027] 1. Cut a nitrocellulose membrane with a pore size of 0.2 microns (produced by Bio-Rad) into small pieces of 5mm*5mm;

[0028] 2. Prepare coating buffer: 50mM / L Tris-Hcl buffer solution with pH 7.8; anti-NSE antibody 10-7938 (Fitzgerald Company), anti-pro-GRP antibody H00002922-M03 ( Abnova), anti-CA19-9 antibody ab3982 (Abcam), anti-CEA antibody MAM02-008 (Meridian), anti-CYFRA21-1 antibody M01300M (Meridian), anti-SCC antibody H00006317M01 (Abnova), human serum albumin (Negative control) Dilute into solution according to the concentration of 0.7-4mg / L;

[0029] 3. Membrane spotting: use BioDot’s BiojetPlus sample spotting system to spot the antibody solution diluted in step 2 at 0.1-2 microliters per spot on the cut nitrocellulose membrane, 56 spots per square centimeter, The dot matrix is ​​arranged as fi...

Embodiment 2

[0044] Embodiment 2 The method for comprehensively detecting lung cancer markers using the kit of the present invention, the specific steps are:

[0045]1. Paste the prepared chip to the bottom of the polystyrene cup with double-sided adhesive;

[0046] 2. Dilute the mixture of SCC, CEA, CA19-9, NSE, CYFRA21-1, and pro-GRP standard substances as quality control substances into a series of gradient concentration mixtures; SCC was purchased from abnova, CEA was purchased from abnova, and CA19- 9 was purchased from Acris, CYFRA21-1 was purchased from Meridian, NSE was purchased from Fitzgerald, and pro-GRP was purchased from abnova;

[0047] 3. Add 100 microliters of patient sample serum and 100 microliters of quality control mixtures of different concentrations into different small cups pasted with chips, shake at 37 degrees, and incubate for 1 hour; wash with washing solution for 6 times ;

[0048] 4. Add 100 microliters of reagent, which contains 15ng of 10-7936, 80ng of PAB...

Embodiment 3

[0054] Example 3 Quality Assessment of the Protein Chip Kit for Comprehensive Detection of Lung Cancer Markers of the Present Invention

[0055] 1. Use diluent to dilute the mixture of standard substances SCC, CEA, CA19-9, CYFRA21-1, NSE, pro-GRP into a series of gradient concentrations;

[0056] 2. Then compare the index SCC with the corresponding detection kit of Shenzhen New Industry Biomedical Engineering Company; compare the index CEA with the corresponding detection kit of Abbott Company; compare the index CA19-9 with the corresponding detection of Shenzhen New Industry Biomedical Engineering Company The kits were compared; the index CYFRA21-1 was compared with the corresponding detection kit of Roche; the index NSE was compared with the corresponding detection kit of Roche; the index pro-GRP was compared with the corresponding detection kit of CanAg; each Indicators were tested at 40 different concentrations;

[0057] 3. Do a correlation analysis between the test resul...

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Abstract

The invention belongs to the technical field of biology and relates to a protein chip reagent kit and a method for comprehensively detecting a lung cancer marker. The protein chip reagent kit for comprehensively detecting the lung cancer marker comprises: (1) a chip (1) and (2) a reaction agent and a detection agent, wherein a plurality of specific antibodies are simultaneously fixed on the chip, can generate antibody-antigen reaction with the lung cancer marker and are fixed on a bottom film of the chip to form a plurality of independent recognition sites; and the reaction agent and the detection agent are used for detecting whether a matter capable of generating antibody-antigen reaction with the specific antibodies exists in a sample to be detected or not through a TRFIA (Time Resolved Fluorescence Immunoassay) method. The reagent kit and the method have the beneficial effects that the six indexes of NSE (Neuron Specific Enolase), SCC (Squamous Cell Carcinoma), CEA (Carcino Embryonie Antigen), CA (Carbonic Anhydrase) 19-9, CYFR (Cytokeratin Fragment) A21-1 and pro-GRP (Glass Reinforced Plastic) can be simultaneously detected, the detection accuracy is improved, the repetitive experimental steps are reduced, and the time and the cost are saved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a protein chip kit and method for comprehensively detecting lung cancer markers. Background technique [0002] Biochip technology is a biotechnology that has emerged in the past ten years. Because it can analyze multiple information in parallel in one reaction, it has been rapidly applied in many biological fields. Biochip technology is classified according to the detection object, mainly divided into two types: gene chip and protein chip. Protein chip is a cutting-edge technology, which directly detects specific functional substances through a microarray composed of various proteins immobilized on a support medium. Existing methods for detecting urine, serum and other body fluids, such as ELISA, radioisotope labeling, gold standard, fluorescence, chemiluminescence, time-resolved fluorescence, etc., are mostly single-index detection with little information. Compared wit...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N21/64
Inventor 王嘎程自卿
Owner 河南生生医疗器械有限公司
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