Cholera morbus toxin detection immunosensor and its preparation method and uses

An immunosensor and cholera toxin technology, applied in the field of sensors, can solve the problems of high instrument cost, unsuitable for rapid detection of Vibrio cholerae, complicated steps, etc., and achieve the effects of simple detection steps, inhibition of non-specific adsorption, and obvious response effect.

Inactive Publication Date: 2008-03-26
HUNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of these detection methods have cumbersome steps, the time required for detection is more than 5 hours, and the cost of the required equipment is high.
Not suitable for rapid detection of Vibrio cholerae at the outbreak site

Method used

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  • Cholera morbus toxin detection immunosensor and its preparation method and uses

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Effect test

Embodiment 1

[0016] Example 1: Preparation of Immunosensor for Cholera Toxin Detection

[0017] 1. Use the newly prepared Piranha lotion (V H2SO4 :V H2O2 =7:3) three times for three minutes each time to clean the surface of the crystal oscillator, then rinsed with pure water, and dried under nitrogen for later use.

[0018] 2. Immerse the treated quartz crystal oscillator in 2mM ethanol solution of octadecyl mercaptan and assemble overnight, so as to self-assemble a uniform and dense hydrophobic layer on the surface of the gold plate of the quartz crystal oscillator;

[0019] 3. Preparation of liposome solution with surface-modified GM1: Weigh 2 mg of DPPC and 0.247 mg of GM1 respectively, dissolve in a mixed solution of methanol and chloroform (1:6 v / v), vacuum rotary evaporation, and remove the organic solvent Leave a layer of uniform phospholipid film on the wall of the flask, add 4mL of phosphate buffer solution (pH=7.4), swell the phospholipid film in a water bath at 50°C for half a...

Embodiment 2

[0021] Embodiment 2: the detection of cholera vaccine

[0022] Cholera toxin detection immunosensor of the present invention is immersed in the phosphate buffer solution (1 / 15M Na 2 HPO 4 and 1 / 15M of KH 2 PO 4 According to the volume ratio of 4:1 mixed preparation, in PBS, pH=7.4), and its frequency was detected by piezoelectric instrument. Under the action of the output voltage of the piezoelectric instrument (-10V ~ 10V), the electronic counter can display the response frequency generated by the AT-cut 9M quartz crystal. By connecting with a computer and data processing software, reading a frequency response data per second, and plotting its frequency response and time can obtain a real-time response curve of the frequency response. When the frequency stabilized, 3 μL of cholera vaccine solution with a final concentration of 1 μg / mL of cholera toxin vaccine provided by Shanghai Liancell Bioengineering Co., Ltd. was added to the detection solution. Real-time detection wa...

Embodiment 3

[0023] Embodiment 3: the detection of blood sample of cholera patient

[0024] The cholera toxin detection immunosensor of the present invention is immersed in the liposome-dispersed phosphate buffer solution with a phospholipid concentration of 0.5 mg / mL, and its frequency is detected by a piezoelectric instrument. Under the action of the output voltage of the piezoelectric instrument (-10V ~ 10V), the electronic counter can display the response frequency generated by the AT-cut 9M quartz crystal. By connecting with a computer and data processing software, reading a frequency response data per second, and plotting its frequency response and time can obtain a real-time response curve of the frequency response. When the frequency of the piezoelectric instrument stabilized, 3 μL of plasma sample was added, which was provided by the Hunan University Hospital. It was observed by real-time detection that the frequency dropped very rapidly at the initial stage, and 90% of the reacti...

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Abstract

This invention provides a kind of cholera toxin detection immunity sensor, includes the quartz crystal shake, the lyophobic layer in the quartz crystal shake and the phosphatides film in the lyophobic layer; this invention also gives the preparation method of the said cholera toxin detection immunity sensor. The preparation of the cholera toxin detection immunity sensor is simple, the reaction is sensitive, the detection is fast, the cost is small and it can be used again and again.

Description

technical field [0001] The invention relates to a sensor, in particular to an immunosensor for detecting cholera toxin, a preparation method and application thereof. Background technique [0002] There have been seven worldwide cholera pandemics in human history. Cholera has become endemic in parts of South Asia, Africa and Latin America, with seasonal outbreaks common and strongly associated with poverty and poor sanitation. Therefore, the timely detection of cholera toxin is of great significance to human beings. The detection methods used now are: ELISA, PCR method, sorbitol fermentation test and CT plate assay, etc. Most of these detection methods have cumbersome steps, the time required for detection is more than 5 hours, and the required equipment costs are high. It is not suitable for the rapid detection of Vibrio cholerae at the outbreak site. The liposome-based cholera toxin immunosensor can detect cholera toxin more quickly and accurately, and the instruments r...

Claims

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Application Information

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IPC IPC(8): G01N33/53
Inventor 蒋健晖吴海龙俞汝勤沈国励陈欢
Owner HUNAN UNIV
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