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Biochip for detecting h5 subtype avian influenza virus and its preparation method and application

A bird flu virus and biochip technology, applied in the direction of measuring devices, instruments, and polarization-influenced characteristics, can solve the problems of low sensitivity, high cost, and long processing cycle, and achieve the effect of low cost

Inactive Publication Date: 2011-12-14
INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Each diagnostic method has its own advantages, but it is limited by several unfavorable factors, such as high cost, long processing cycle and low sensitivity, especially in the quantitative analysis of avian influenza virus content and the immediate and accurate determination of virus subtypes. needs improvement

Method used

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  • Biochip for detecting h5 subtype avian influenza virus and its preparation method and application
  • Biochip for detecting h5 subtype avian influenza virus and its preparation method and application
  • Biochip for detecting h5 subtype avian influenza virus and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1 : Preparation of biochip

[0076] This example is to prepare the biochip for detecting H5 subtype avian influenza virus provided by the present invention, including first preparing the gold film on the surface of the chip, then modifying the gold film with carboxymethyl dextran, and finally immobilizing the anti-H5N1 Monoclonal Antibody to Subtype Avian Influenza Virus. The specific steps are detailed as follows:

[0077] (1) get rid of with uniform photoresist on the glass sheet, remove the photoresist in the area that needs to cover gold film on the glass sheet surface with photolithography;

[0078] (2) Adopting the method of vacuum evaporation coating, first deposit about 2nm of chromium on a 18mm×18mm×0.14mm glass plate, and then deposit 50nm of gold with a purity of 99.9999%. Before the gold film is officially used, use the newly prepared piranha solution (30% H 2 o 2 :H 2 SO 4 =1:3; v / v) for 2 minutes, or use an ion etching machine to treat the ...

Embodiment 2

[0083] Example 2 : How to use the biochip

[0084] This example is the method of using the biochip prepared in Example 1, which is described in detail as follows. Among them, the system model diagram of the surface plasmon resonance biosensor antibody chip is as follows figure 1 shown.

[0085] 1. Processing of samples to be tested:

[0086] (1) Tissue sample: Take the tissue sample to be tested and put it in a grinder to cut it into pieces and grind it, add an appropriate amount of PBS buffer solution and continue grinding until it is in a homogenized state, then take the homogenate solution and sample processing buffer (the composition is as follows: NaCl 137mmol / L, KCl 2.7mmol / L, Na 2 HPO 4 4.3mmol / L, KH 2 PO 4 1.4mmol / L and 2% Triton X-100, pH 7.2) mixed according to the volume ratio of 1:1, left at room temperature for more than 90 minutes, then it can be used for detection;

[0087] (2) Inactivated secretions and excreta samples to be tested: fully twist the c...

Embodiment 3

[0096] Example 3 : Application of biochip

[0097] This example uses the biochip prepared in Example 1 to detect the content of the H5N1 subtype avian influenza virus in the allantoic fluid of the H5N1 subtype avian influenza virus, and the specific steps are described in detail as follows.

[0098] (1) Chicken embryo allantoic fluid sample processing: Take H5N1 virus (A / plateau pika / Qinghai / 04 / 2007) chicken embryo allantoic fluid and chicken embryo allantoic fluid blank control respectively with sample processing buffer in a volume ratio of 1: 1 Mix and place at room temperature for more than 90 minutes, then it can be used for detection;

[0099] (2) Install the detection biochip on the SPR-2002 biosensor prism according to the operation method of the instrument instruction manual, wherein the basic structural model diagram of the sensor is as follows figure 2 shown;

[0100] (3) Dilute the sample treatment solution 1000 times with phosphate buffer solution to obtain th...

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Abstract

The invention provides an antibody biochip for detecting H5 subtype avian influenza virus, its preparation method and application. The biochip provided by the present invention comprises a solid phase carrier, a gold film attached to the solid phase carrier, and an anti-H5N1 subtype avian influenza virus monoclonal antibody connected with the gold film, and the biochip is helpful for the detection of the H5 subtype The rapid and accurate detection and identification of avian influenza virus, and the quantitative analysis of H5 subtype avian influenza virus in the sample can truly reflect the advantages of specificity, sensitivity and rapidity of the biochip.

Description

technical field [0001] The invention belongs to the field of biochips, and relates to a biochip for detecting and analyzing H5 subtype avian influenza virus, a preparation method and application thereof. Background technique [0002] Biochips have shown broad application prospects in various fields such as bacterial identification, environmental monitoring, and pathogen detection. Compared with traditional instrument detection methods, biochips are characterized by real-time, label-free, small sample volume, high-throughput, miniaturization, automation, low cost, pollution prevention, and fast detection speed. They are widely used in clinical diagnosis and virus detection. has great advantages. [0003] At present, although chip technology has made great progress, there are still many problems in sample preparation, probe immobilization, molecular labeling, etc., and foreign related detection instruments and chips are expensive, which seriously restricts domestic chip techn...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/552G01N33/569G01N21/21
Inventor 何宏轩崔大付王海静李辉张璐璐董国英
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
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