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Manufacturing method of schistosome high sensitivity gas phase piezoelectric immunosensor and silver-depositing high sensitivity gas phase piezoelectric immunol sensing detection method

An immunosensor and schistosomiasis technology, which is applied in the field of clinical medical identification and detection, can solve the problems of low detection sensitivity, difficulty in realizing large-scale sample detection, etc., and achieves simple, fast and convenient batch detection of devices and preparation processes, and realizes integration. Effect

Inactive Publication Date: 2010-06-16
HUNAN UNIV
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AI Technical Summary

Problems solved by technology

[0008] The technical problem to be solved by the present invention is to overcome the limitation that the detection sensitivity of the existing gas-phase piezoelectric immunosensing detection technology is not high and the existing enzyme-catalyzed mass amplification liquid-phase piezoelectric immunosensing detection technology is difficult to realize the detection of large batches of samples Combining the simple and fast gas-phase piezoelectric sensing technology and the amplification effect of enzyme-catalyzed metallic silver deposition, a preparation method of a highly sensitive gas-phase piezoelectric immunosensor for schistosomiasis and a highly sensitive gas-phase piezoelectric immunosensing detection method of metallic silver deposition are provided. The electroimmunosensor has a simple preparation process and low cost, and the high-sensitivity gas-phase piezoelectric immunosensor detection technology can realize early batch detection of schistosomiasis

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0026] Example 1: Preparation of a highly sensitive mass-amplified schistosoma gas-phase piezoelectric immunosensor:

[0027] (1) Pretreatment of crystal oscillator: use the newly prepared piranha washing solution (concentrated H 2 SO 4 : H 2 O 2 =3:1 (v / v)) washed three times for 3 minutes each time to obtain a clean crystal oscillator, rinsed with ethanol and deionized water, and dried under nitrogen;

[0028] (2) Antigen immobilization: soak the pretreated crystal oscillator in cysteamine solution (10 mmol / L) overnight at room temperature, rinse with ethanol; add a certain amount of self-assembled cysteamine to the quartz crystal oscillator dropwise 2.5% glutaraldehyde solution, placed at room temperature for 1 hour, washed with deionized water and dried, then 20 μL of 10 μg / mL antigen SjAg32 was added dropwise to the crystal, and placed at room temperature for 1 hour;

[0029] (3) Blocking: After washing the antigen-immobilized quartz crystal oscillator with phosphate ...

Embodiment 2

[0031] Immerse the prepared piezoelectric quartz crystal oscillator in 0.1-1000ng / mL schistosome antibody solution, incubate at 37°C for 1 hour, and then wash with PBS containing 0.1% sodium chloride under magnetic stirring for 10 minutes to remove unbound antibodies. Antibody; then add 40 μL ALP-goat anti-rabbit IgG (0.1 mg / mL) dropwise to the crystal oscillator, incubate at 37°C for 1 hour, and place the quartz crystal oscillator after washing with PBS in a closed detection air chamber, connect the quartz crystal oscillator frequency The monitoring instrument is used for testing, and when the frequency is stable, record the initial frequency value F 0 , and then add 30 μL of phosphorylated ascorbate and AgNO dropwise on the crystal oscillator 3 (1mmol / L), mix the solution, incubate at 37°C for 1 hour, add 50 μL of thiourea (10mmol / L) dropwise to the crystal after the reaction, wash off after 3 minutes, dry for 1 hour, and then detect the frequency value F '. The sensor can...

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Abstract

The invention provides a silver-depositing high sensitivity gas phase piezoelectric immunol sensing technique and an immunol sensing method for detecting Japanese schistosome. The manufacture of an immunosensor means the assembly of the immunosensor, and the immunosensor is formed by piezoelectric quartz crystal oscillation pretreatment, antigen fossilization and sealing processes. The antibody immunol sensing detection method comprises an immunoreaction process and a detection process. The gas phase piezoelectric immunol sensing technique is adopted, and enzymes are combined to catalyze silver-depositing effect so that the mass-basis response of sensor interfaces is increased, the sensitivity of the sensor can be improved and a plurality of samples can be simultaneously detected. The high sensitivity gas phase piezoelectric immunol sensing technique which is based on silver-depositing has the advantages of simple equipment and low cost, and can realize the batch detection of immune molecules.

Description

technical field [0001] The invention relates to the technical field of clinical medical identification and detection, and further relates to a preparation method of a schistosomiasis highly sensitive gas-phase piezoelectric immunosensor and a metal silver deposition high-sensitivity gas-phase piezoelectric immunosensing detection method. Background technique [0002] In recent years, piezoelectric immunosensors have received extensive attention in the field of immunodetection. Piezoelectric immunosensing technology is a biosensing technology that combines the high-sensitivity piezoelectric effect of quartz crystal and the highly specific recognition function of immune response. and other parameters. This technology has many advantages of an ideal biosensing technology, including simplicity, sensitivity, rapidity, wide response range, cheap instruments, small size, easy integration and online automated detection, etc. It is used for biochemical analysis without tracer labels...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
Inventor 吴朝阳周瑾艳申田田沈国励俞汝勤
Owner HUNAN UNIV
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