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PGI rapid quantitative detection kit and making method and detection method thereof

A quantitative detection and production method technology, applied in the biological field, can solve the problems of easy detachment and instability of molecules, and achieve the effect of high efficiency and short detection time

Inactive Publication Date: 2016-05-04
SHENZHEN BLOT BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the colloidal gold label is based on the principle of electrostatic adsorption, it is unstable in the fluid, and the labeled molecules are easy to fall off, and can only be read when the colloidal gold particles are enriched to a certain amount visible to the naked eye. Qualitative analysis of the detected object
At present, there is no relevant quantum dot immunochromatographic technology to quickly detect PGI

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] A kind of manufacture method of PGI detection test strip, comprises the following steps:

[0033] a. Conjugate the PGI monoclonal antibody with quantum dots to obtain the quantum dot-PGI monoclonal antibody complex, and spray it on the conjugate pad;

[0034] b. Coat the PGI monoclonal antibody on the nitrocellulose membrane as the detection line, and coat the anti-mouse anti-mouse antibody on the nitrocellulose membrane as the quality control line, the distance between the detection line and the quality control line is 5mm.

[0035]c. Lap the sample pad, the conjugate pad sprayed with the quantum dot-PGI monoclonal antibody complex, the nitrocellulose membrane with the detection line and the quality control line, and absorbent paper on the sticky bottom plate in sequence. The test strips that overlap each other by 1mm and are cut into 4mm width after being glued are the PGI test strips.

[0036] Method for preparing quantum dot-PGI monoclonal antibody complexes by co...

Embodiment 2

[0051] Method for preparing quantum dot-PGI monoclonal antibody complexes by coupling PGI monoclonal antibody to quantum dots:

[0052] Take 0.1ml of quantum dots (excitation wavelength 365nm, emission wavelength 620nm) solution composed of CdSe / ZnS, surface group -COOH, put in 1ml of MES buffer solution with a concentration of 0.1mol / LPH5.0, add 0.15mg of PGI Monoclonal antibody, mix well. Add 0.01ml of 50mg / ml 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), mix immediately, and incubate at room temperature for 60min, then add 0.1ml of 1% glycine solution, Incubate at room temperature for 0.5h, centrifuge (23000rpm, 30min), discard the supernatant, add 0.1ml preservation solution (0.02mol / L phosphate buffered saline, containing 1% BSA, 0.04% Proclin300), the obtained quantum dot-PGI monoclonal Antibody complexes were stored at 4°C until use.

[0053] Except that the above coupling method is different from Example 1, other test strip preparation methods are the same as ...

Embodiment 3

[0063] Method for preparing quantum dot-PGI monoclonal antibody complexes by coupling PGI monoclonal antibody to quantum dots:

[0064] Take 0.1ml of quantum dots (excitation wavelength 365nm, emission wavelength 620nm) solution composed of CuInZnS / ZnS, surface group is -COOH, put in 1ml of MES buffer solution with a concentration of 0.1mol / LPH5.0, and add 0.02ml concentration 50mg / ml 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) and 0.08ml of 20mg / ml N-hydroxysulfosuccinimide (sulfo-NHS), incubated at room temperature After 20min, centrifuge (23000rpm, 30min), discard the supernatant, wash to obtain the quantum dot solvent, centrifuge under the same conditions, discard the supernatant to obtain the final quantum dot solvent, add 0.1mg of PGI monoclonal to the final quantum dot solvent Antibody, incubate at room temperature for 2h, add 1% glycine, incubate at room temperature for 0.5h, centrifuge under the same conditions, discard the supernatant and add 0.1ml preservati...

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Abstract

The invention relates to a PGI rapid quantitative detection kit and a making method and detection method thereof. The detection kit comprises a detection card and a sample buffer solution disposed in the kit, the test strip in the detection card comprises a sample pad, a conjugate pad, a nitrocellulose membrane and absorbent paper that are lapped in order. PGI monoclonal antibody coupled quantum dots are sprayed to the conjugate pad, the nitrocellulose membrane is coated with another strain of PGI monoclonal antibody as a detection line, and is also coated with goat-anti-mouse antibody or other anti-mouse antibodies as a quality control line. The kit provided by the invention combines quantum dot and immunochromatography technologies, has the characteristics of short detection time and high efficiency, and can well detect the storage of PGI. The kit and the detection method provided by the invention utilize quantum dot immune chromatography technology, which has the characteristics of short detection time and high efficiency, and can detect the stock of very well. The invention uses quantum dot immunochromatography, can realize non-traumatic, low risk, safe, cheap, accurate and rapid quantitative detection of PGI content in serum, plasma and whole blood, and can provide assistance for stomach disease inspection and disease monitoring.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a PGI rapid quantitative detection kit and its production and detection methods. Background technique [0002] Pepsinogen (Pepsinogen, PG) is the precursor of pepsin, including pepsinogen I (PGI) and pepsinogen II (PGI). PGI is mainly secreted by the principal cells and mucus cells of the gastric body, most of which are secreted into the gastric cavity, and a small amount can enter the blood through some channels. PGI is mainly produced by the principal cells and neck cells of the pyloric gland in the gastric antrum and the Bruner gland mucosa in the proximal duodenum, and a small amount can enter the blood through some channels. When the stomach environment changes, such as pathological changes in the gastric mucosa, the secretion of PGI and PGI will be affected, which in turn will affect the content of PGI and PGI in the blood. Conversely, the content of PGI and PGI in the blood c...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2800/062
Inventor 张永顶马伟民张大准马新民
Owner SHENZHEN BLOT BIOTECH
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