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Combined detection method for pepsinogen I and pepsinogen II

A pepsinogen and combined detection technology, applied in the field of in vitro diagnosis, can solve problems such as high inspection costs, high technical requirements for operators, and radiation exposure, and achieve the effect of avoiding human body damage, avoiding the inconvenience of gastroscopy, and good correlation

Inactive Publication Date: 2019-12-20
NINGBO AUCHEER BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

X-ray, as a traditional detection method for gastric cancer, is more valuable in diagnosis of advanced gastric cancer or diffuse gastric cancer, but this method has problems such as radiation exposure and high inspection costs, and it is powerless for the judgment of early gastric cancer; the cost of endoscopy is relatively high , the technical requirements for the operators are high, and the gastroscopy is very painful for the patients. As a result, many patients are discouraged, and it cannot be used as a general survey method for patients with subclinical symptoms; little diagnostic value
[0004] Compared with other gastric detection methods, the combined detection of serum PGI / PGII has the advantages of non-invasive, simple, reliable and low cost. At present, a variety of detection methods have been established, such as enzyme-linked immunoassay, chemiluminescence method, and immunoturbidimetric method , time resolution, etc., and have been widely used in hospitals and medical examination centers, but often require expensive instruments and reagents

Method used

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  • Combined detection method for pepsinogen I and pepsinogen II
  • Combined detection method for pepsinogen I and pepsinogen II
  • Combined detection method for pepsinogen I and pepsinogen II

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Experimental program
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Embodiment 1

[0023] The combined detection method of pepsinogen Ⅰ and pepsinogen Ⅱ comprises the following steps:

[0024] (1) Immunochromatography test strip assembly: Lap the sample pad, binding pad, reaction membrane and water-absorbent pad sequentially and paste them on the base plate. The thickness of the base plate is 0.2 mm, the length is 7 cm, and the width is 4 mm. A detection line I and a detection line II are also arranged between the line and the bonding pad, and the detection line II is located between the detection line I and the control line,

[0025] (2) Antibody coating: fluorescent latex-labeled pepsinogen Ⅰ monoclonal antibody with a concentration of 0.4 mg / ml, fluorescent latex-labeled pepsinogen Ⅱ monoclonal antibody with a concentration of 0.4 mg / ml and a concentration of 0.4 mg / ml of mouse anti-rabbit IgG coated on the binding pad, the concentration of 0.8mg / ml pepsinogen Ⅰ monoclonal antibody, the concentration of 0.8mg / ml pepsinogen Ⅱ monoclonal antibody and the c...

Embodiment 2

[0029] The combined detection method of pepsinogen Ⅰ and pepsinogen Ⅱ comprises the following steps:

[0030] (1) Immunochromatography test strip assembly: Lap the sample pad, binding pad, reaction membrane and water-absorbent pad sequentially and paste them on the bottom plate. The thickness of the bottom plate is 0.3mm, the length is 6cm, and the width is 4mm. A detection line I and a detection line II are also arranged between the line and the bonding pad, and the detection line II is located between the detection line I and the control line,

[0031] (2) Antibody coating: fluorescent latex-labeled pepsinogen Ⅰ monoclonal antibody with a concentration of 0.4 mg / ml, fluorescent latex-labeled pepsinogen Ⅱ monoclonal antibody with a concentration of 0.4 mg / ml and a concentration of 0.4 mg / ml of mouse anti-rabbit IgG coated on the binding pad, the concentration of 0.8mg / ml pepsinogen Ⅰ monoclonal antibody, the concentration of 0.8mg / ml pepsinogen Ⅱ monoclonal antibody and the ...

Embodiment 3

[0035] The present invention also includes a preparation method of fluorescent latex-labeled pepsinogen I monoclonal antibody:

[0036] Add 2% BSA to a 0.05MPBS solution with a pH value of 7.5 and store at 4°C to prepare a labeled washing solution; add 50mg / ml EDC solution to a 0.05MPBS solution buffer with a pH value of 7.5 to prepare a latex crosslinking agent. Mix 1% BSA, 0.5% skimmed milk, 0.05% NaN3, 0.1% Tween-20 and 0.01MPBS solution with a pH value of 7.5 to prepare a latex preservation solution; After reacting at room temperature for 1 hour, centrifuge for 15 minutes, centrifuge 3 times with 0.05 MPBS to obtain activated fluorescent latex; add 0.4 mg of pepsinogen I monoclonal antibody per ml of activated fluorescent latex, mix and stir at room temperature React for 2-4 hours, centrifuge for 30 minutes, discard the supernatant, wash the precipitate twice with labeled washing solution, discard the supernatant for the last time, dissolve the precipitate with latex prese...

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Abstract

The invention discloses a combined detection method for pepsinogen I and pepsinogen II. The method comprises the steps of immunochromatography test strip assembly, antibody coating, sample diluent preparation, sample detection and the like. The combined detection method for pepsinogen I and pepsinogen II utilizes a fluorescence immunochromatographic technique and adopts a double-antibody sandwichmethod, can detect the content of pepsinogen I / II in serum or plasma 10 minutes after sample adding, has the advantages of simplicity, convenience and rapidness, and avoids the damage of X-ray to human body and the inconvenience of gastroscope; and meanwhile, through the detection of samples with different concentrations, the sensitivity is higher, and the correlation is good.

Description

technical field [0001] The invention relates to the field of in vitro diagnosis, in particular to a combined detection method for pepsinogen I and pepsinogen II. Background technique [0002] Pepsinogen (PG) is the precursor of pepsin. According to its biochemical properties and immunogenicity, it is divided into 2 subgroups. Components 1-5 have the same immunogenicity and are called pepsinogen I. Secreted by chief cells and mucus neck cells of fundic glands; Component 6-7 is called pepsinogen II, except secreted by chief cells and mucus neck cells of fundic glands, mucus necks of cardiac glands and pyloric glands of gastric antrum Cells as well as the upper duodenum also produce pepsinogen II. The level of serum pepsinogen reflects the morphology and function of gastric mucosa in different parts: PGI is a pointer to detect the function of gastric acid gland cells. When gastric acid secretion increases, PGI increases, and when secretion decreases or gastric mucosal glands a...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/573G01N33/558
CPCG01N33/558G01N33/573G01N33/577
Inventor 唐静崔丛丛董文霞周义正
Owner NINGBO AUCHEER BIOTECHNOLOGY CO LTD
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