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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Problems solved by technology

Most of these detection methods have cumbersome steps, the time required for detection is more than 5 hours, and t

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

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Example Embodiment

[0016] Example 1: Preparation of cholera toxin detection immunosensor

[0017] 1. The new Piranha lotion (V H2SO4 : V H2O2 =7:3) Wash three times for three minutes each time to clean the surface of the crystal oscillator, then wash with pure water, and dry under nitrogen for later use.

[0018] 2. Immerse the treated quartz crystal 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 disk of the quartz crystal;

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

Example Embodiment

[0021] Example 2: Detection of cholera vaccine

[0022] The cholera toxin detection immunosensor of the present invention was immersed in a liposome-dispersed phosphate buffer solution with a phospholipid concentration of 0.5 mg / mL (1 / 15 M Na 2 HPO 4 and 1 / 15M of KH 2 PO 4 According to the volume ratio of 4:1, the mixture was prepared in PBS, pH=7.4), and the frequency was 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 to a computer and data processing software, reading one frequency response data per second, and plotting its frequency response against time, the real-time response curve of the frequency response can be obtained. When the frequency stabilized, 3 μL of cholera toxin vaccine solution with a final concentration of 1 μg / mL provided by Shanghai United Cell Bioengineering C...

Example Embodiment

[0023] Example 3: Detection of blood samples from cholera patients

[0024] The cholera toxin detection immunosensor of the present invention is immersed in a phosphate buffer solution dispersed in liposomes 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 to a computer and data processing software, reading one frequency response data per second, and plotting its frequency response against time, the real-time response curve of the frequency response can be obtained. When the frequency of the piezoelectric instrument stabilizes, add 3 μL of plasma sample, which is provided by Hunan University Hospital. Using real-time detection, it is observed that the frequency drops very rapidly at the initial stage, and 90% of the respo...

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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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