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A kind of electrochemical detection method of chicken infectious bronchitis virus h120 strain

A bronchitis and chicken infectious technology, applied in the field of analytical chemistry, can solve the problems of expensive instruments, endangering the life and health of animals, and expensive, and achieve the effect of avoiding the degradation of RNA

Active Publication Date: 2022-04-12
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These assays often require expensive instrumentation, or require radioactive or enzymatic labeling of the catalytic substrate
As we all know, some animal experiments are very time-consuming and expensive, and may even endanger animal life and health

Method used

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  • A kind of electrochemical detection method of chicken infectious bronchitis virus h120 strain
  • A kind of electrochemical detection method of chicken infectious bronchitis virus h120 strain
  • A kind of electrochemical detection method of chicken infectious bronchitis virus h120 strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1IB

[0030] Example 1. Feasibility verification of electrochemical detection method for IBV H120 strain

[0031] Under the optimal experimental conditions, the system specificity verification was carried out. According to the above method, the IBV NNA strain RNA stock solution and RNase-free double distilled water were used to replace the IBV H120 strain RNA stock solution in step (5) to verify the feasibility of the detection system. Specifically, take IBV H120 strain RNA stock solution, IBVNNA strain RNA stock solution and RNase Free ddH 2 Add 2 μL of 10 μM H120-target and 85 μL of Hybridization Buffer (10 mM PBS, 0.25M NaCl, pH=7.0) to 2 μL each, mix well, and perform the experiment according to the above experimental steps (1) to (7). Electrochemical scans were performed under optimal experimental conditions. The result is as figure 2 As shown in A-B, the detection system detects the H120 strain, but cannot detect the NNA strain, which proves that the detection system is fe...

Embodiment 2

[0032] Example 2. Determination of the electrochemical signal-concentration logarithmic standard curve of different concentrations of virus liquid

[0033]The preparation concentration in the above-mentioned method step (5) is respectively 1.56e -9 μM, 1.56e -8 μM, 1.56e -7 μM, 1.56e -6 μM H120 virus sample, the prepared sample was scanned by linear voltammetry under the optimal experimental conditions. The result is as image 3 As shown in A, when the virus concentration increases, the peak current value increases. Virus fluid at 1.56e -9 μM-1.56e -6 In the range of μM, the logarithm and LSV of the virus concentration 500 The current value (ip) of mV has a good linear relationship ( image 3 B), both meet y=7.9821+0.6406lgx (R 2 =0.99837), where y is the peak value of the electrochemical signal (μA), and x is the concentration (μM) of the H120 strain virus liquid, which can be used as the basis for quantitative detection of the IBV H120 strain in the actual sample. ...

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Abstract

The invention belongs to the technical field of analytical chemistry, and relates to an electrochemical detection method and application of chicken infectious bronchitis virus H120 strain. The present invention converts the RNA of chicken infectious bronchitis virus H120 strain into H120-target which can specifically recognize the RNA of H120 strain by means of annealing, S1 nuclease (S1) digestion and high temperature heating, based on self-assembly The sulfhydryl-modified nucleic acid probe 2 was immobilized on the surface of the gold electrode and incubated with H120-target, and the gold nanoparticles modified by the nucleic acid probe 1 that could specifically bind to the H120-target were introduced to adsorb the electrical signal molecule ruthenium hexaamine (RuHex ) is used to generate and amplify the signal, and finally the adsorbed RuHex is detected electrochemically by linear voltammetry, and the linear equation is obtained by drawing the standard relationship curve between the H120 strain concentration and the electrical signal, and the actual value is calculated according to the detected electrical signal. The content of the H120 strain in the samples. The method has the advantages of low cost, high sensitivity and strong specificity.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry and relates to an electrochemical method for detecting chicken infectious bronchitis virus H120 strain. Background technique [0002] Chicken infectious bronchitis is an acute and highly contagious infectious disease caused by coronavirus genus Infectious Bronchitis Virus (IBV), which has caused huge economic losses to the chicken industry worldwide. IBV is a single-stranded virus whose genes can be mutated by point mutation and recombination. There are many serotypes of IBV, and there is no cross-reaction or little cross-reaction between different serotypes, which increases the difficulty of immune prevention. Therefore, it is of great practical significance to establish an effective method to detect different strains of IBV to assist in the selection of a suitable vaccine. [0003] Nanotechnology has become a hot spot in the field of material research today. Gold nanoparticles are...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/48
CPCG01N27/3276G01N27/3278G01N27/48
Inventor 许媛媛苗晋锋吴荣春
Owner NANJING AGRICULTURAL UNIVERSITY
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