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Hepatitis C virus variation detection protein chip, preparation method and application thereof

A hepatitis C virus and protein chip technology, applied in the field of hepatitis virus detection protein chip, can solve the problems of lack of clinical significance, expensive laser confocal scanner, and no visible protein chip for detection of hepatitis C virus, etc. good sex effect

Inactive Publication Date: 2010-01-06
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this product has not been widely used in my country, and the need to purchase expensive laser confocal scanners and the lack of clear clinical significance are the main reasons for limiting its promotion.
At present, there is no report on the visualization protein chip for detection of hepatitis C virus

Method used

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  • Hepatitis C virus variation detection protein chip, preparation method and application thereof
  • Hepatitis C virus variation detection protein chip, preparation method and application thereof
  • Hepatitis C virus variation detection protein chip, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 HCV mutation detection protein chip reaction matrix design and protein chip preparation

[0030]1. Materials: anion exchange column Q-Sepharose FF, gel filtration column Sephardex G-50 were purchased from Pharmacia Biotech; urea and sucrose were purchased from Beijing Chemical Reagent Company; lysozyme and BSA were purchased from Promega; aminosilane The chemical glass slides were purchased from Beijing Boao Biochip Co., Ltd.; the ultrasonic crushing instrument was purchased from Ningbo Xinzhi Technology Co., Ltd.

[0031] 2. Method results:

[0032] 1. We designed a 6×6 reaction matrix consisting of 16 multi-target HVR1 antigens. Each antigen was replicated vertically twice, and a positive IgG control and an IL1 negative control were set up (attached figure 1 ).

[0033] 2. Cultivation of expression strains: Take 20 μl of expression strains stored at -70°C (preserved in this room. The first hypervariable region antigen of hepatitis C virus and its use in th...

Embodiment 2

[0040] Example 2 HCV mutation detection protein chip reaction and visualization of results

[0041] 1. Materials: Colloidal gold-labeled protein A: purchased from Beijing Zhongshan Biotechnology Company; hydroquinone sodium citrate, citric acid, and silver nitrate were purchased from Beijing Chemical Reagent Company;

[0042] 2. Method results:

[0043] Take the chips out of the refrigerator and rewarm for 20 minutes, wash the chips with pure water, and suck out excess water. The serum was diluted 10 times with the chip diluent, dropped onto the chip reaction area, and kept at room temperature for 1 hour. Wash the slices 5 times with washing solution, wash the slices with pure water and suck out excess water. Add colloidal gold-labeled protein A (diluted with PBS 1:100), react at room temperature for 10 minutes, wash the slices with washing solution 5 times, wash the slices with pure water, suck out excess water, add silver chromogenic solution (solution A: terephthalate Di...

Embodiment 3

[0044] Example 3 Clinical significance of visualized HCV mutation detection protein chip

[0045] 1. Materials: 28 serum samples of asymptomatic HCV infection were provided by Beijing Red Cross Blood Center; 41 serum samples of patients with chronic hepatitis C and 37 serum samples of patients with liver cirrhosis were provided by People's Hospital Affiliated to Peking University; serum samples before and after interferon treatment for hepatitis C were provided by Provided by the Second Affiliated Hospital of Chongqing Medical University;

[0046] 2. Method results:

[0047] 1. Visualize the relationship between HCV mutation detection protein chip and disease chronicity process

[0048] The heterogeneity of HVR1 antibody in 28 sera of asymptomatic HCV patients, 41 sera of chronic patients and 37 sera of patients with liver cirrhosis was detected by visual HCV mutation detection protein chip. The result is as image 3 Shown: The number of positive multi-target HVR1 antibodie...

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Abstract

The invention discloses a Hepatitis C virus variation detection protein chip as well as a preparation method and application thereof. The invention mainly adopts colloid gold-silver dye detection system to replace original fluorescent marker of the chip, with the application of the detection system of the invention, a chip fluorescent scanner is unnecessary, only by visual detection and simple image scanning can the reaction result of the chip be judged, thus realizing the aim of HCV variation detection. Compared with the fluorescent detection system, the invention is more suitable for fundamental medical treatment organs, can be applied to prognosis of HCV diseases, prediction of curing effect of interferon and the like, and has certain clinical application prospect.

Description

technical field [0001] The invention relates to a protein chip for detecting hepatitis C virus, in particular to a protein chip for detecting hepatitis C virus, and also relates to the medical application of the chip. Background technique [0002] Biochip technology is one of the most far-reaching major scientific and technological advances since the mid-1990s. It is a highly intersecting new technology integrating microelectronics, biology, physics, chemistry, and computer science. It has great basic research value. It also has obvious industrialization prospects. At present, according to the different types of recognition molecules fixed on the substrate, biochips have developed various types such as gene chips, protein chips, and tissue chips, among which protein chips are a new technology born in response to the "post-genome era". And it is widely used in many aspects such as disease diagnosis, drug screening, and personalized drug delivery, bringing profound and extens...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/576
Inventor 张贺秋修冰水王国华柴文菡凌世淦宋晓国
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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