Fast and quantitative PG (pepsinogen)II detection kit, production method thereof and PGII detection method

A technology for quantitative detection and production methods, which is applied in measurement devices, instruments, disease diagnosis, etc., can solve the problems of easy detachment and instability of molecules, and achieves the effect of high efficiency and short detection time.

Inactive Publication Date: 2016-04-06
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 immunochromatography technology to quickly detect PGⅡ

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A kind of manufacture method of PGⅡ detection test paper, comprises the following steps:

[0031] a. Use quantum dots to couple PGⅡ monoclonal antibody to obtain quantum dot-PGⅡ monoclonal antibody complex, and spray it on the conjugate pad;

[0032] b. The PGⅡ monoclonal antibody was coated on the nitrocellulose membrane as the detection line, and the anti-mouse anti-mouse antibody was coated on the nitrocellulose membrane as the quality control line, the distance between the detection line and the quality control line is 5mm.

[0033] c. Lap the sample pad, the conjugate pad sprayed with the quantum dot-PGⅡ 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 cut into 4mm width after being glued are the PGⅡ test strips.

[0034] Method for preparing quantum dot-PGII monoclonal antibody complexes by cou...

Embodiment 2

[0049] Method for preparing quantum dot-PGII monoclonal antibody complexes by coupling PGⅡ monoclonal antibody to quantum dots:

[0050] Take 0.1ml of quantum dots (excitation wavelength 365nm, emission wavelength 620nm) composed of CdSe / ZnS and surface group -COOH, and put them in 1ml of MES buffer with a concentration of 0.1mol / LPH5.0, add 0.15mg of PGⅡ 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-PGⅡ monoclonal Antibody complexes were stored at 4°C until use.

[0051] Except that the above coupling method is different from Example 1, other test strip preparation methods are the same as Examp...

Embodiment 3

[0061] Method for preparing quantum dot-PGII monoclonal antibody complexes by coupling PGⅡ monoclonal antibody to quantum dots:

[0062] 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 PGⅡ 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 preservat...

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Abstract

The invention relates to a fast and quantitative PG (pepsinogen)II detection kit, a production method thereof and a PGII detection method. The detection kit comprises a detection card and a sample buffer which are arranged in the kit, wherein a test strip in the detection card comprises a sample pad, a conjugate pad, a nitrocellulose membrane and absorbent paper which are in sequential lap joint; quantum dots coupled with one PGII monoclonal antibody are sprayed to the conjugate pad, and the nitrocellulose membrane is coated with another PGII monoclonal antibody as a detection line and is further coated with a goat-anti-mouse antibody or another anti-mouse antibody as a quality control line. According to the detection kit, the quantum dots are combined with immunochromatography, the detection kit has the characteristics of short detection time and high efficiency and can detect the stock of PGII very well. A quantum dot-immunochromatographic method is adopted, the content of the PGII in serum, plasma and whole blood is detected non-invasively, safely, accurately, fast and quantitatively with low risk and low cost, and an auxiliary function can be provided for gastric disease examination and monitoring.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a PGII 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 (PGII). 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. PGII 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 gastric environment changes, such as pathological changes in the gastric mucosa, the secretion of PGI and PGⅡ will be affected, which in turn will affect the content of PGI and PGⅡ in the blood. Conversely, the content of PGI and PGⅡ in the bloo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573G01N33/577G01N33/558G01N33/533
CPCG01N33/573G01N33/533G01N33/558G01N33/57446G01N33/577G01N2800/06
Inventor 张永顶马伟民张大准马新民
Owner SHENZHEN BLOT BIOTECH
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