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Pepsinogen I (PGI) detection kit and detection method thereof

A pepsinogen and detection kit technology, applied in the field of biomedicine, can solve the problems of expensive equipment, poor stability, etc., and achieve the effects of reducing the amount of detection samples, increasing stability, and improving work efficiency

Inactive Publication Date: 2018-08-14
广州市伊川生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method can be automated and has high accuracy, but the equipment is expensive and the stability is poor

Method used

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  • Pepsinogen I (PGI) detection kit and detection method thereof
  • Pepsinogen I (PGI) detection kit and detection method thereof
  • Pepsinogen I (PGI) detection kit and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Pepsinogen I (PGI) detection kit and detection method thereof, adding glycerol stabilizer:

[0034] R 1 : Phosphate buffer 100mmoL / L, Proclinc 3000.5g / L, polyethylene glycol 600010g / L, Triton 1001g / L, TRIS 10g / L;

[0035] R 2 : Phosphate buffer 100mmoL / L, latex particle anti-human PGI antibody-conjugated emulsion (anti-human pepsinogen I (mouse) monoclonal antibody sensitive emulsion solution 1.3mg / mL), Proclinc 3000.5g / L, calf serum Albumin (BSA) 10g / L, glycerol 90g / L;

[0036] Wherein, the preparation method of latex particle anti-human PGI antibody binding emulsion (anti-human pepsinogen I (mouse) monoclonal antibody sensitive emulsion solution) is:

[0037] Step 1: Take 2mL of carboxylated latex microspheres (150nm, 5% w / v) and add phosphate buffer to 4ml;

[0038]Step 2: Add 50ml 0.2g / L of N-hydroxysuccinimide (NHS) and 50ml 0.2g / L of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride Salt (EDC) for activation, stir well and incubate at 37°C for 20min...

Embodiment 2

[0043] Pepsinogen I (PGI) detection kit and its detection method, without glycerol:

[0044] R 1 : Phosphate buffer 100mmoL / L, Proclinc 3000.5g / L, polyethylene glycol 600010g / L, Triton 1001g / L, TRIS 10g / L;

[0045] R 2 : Phosphate buffer 100mmoL / L, latex particle anti-human PGI antibody conjugated emulsion (anti-human pepsinogen I (mouse) monoclonal antibody sensitive emulsion solution: 1.3mg / mL), Proclinc 3000.5g / L, calf Serum albumin (BSA) 10g / L;

[0046] Wherein, the preparation method of latex particle anti-human PGI antibody binding emulsion (anti-human pepsinogen I (mouse) monoclonal antibody sensitive emulsion solution) is:

[0047] Step 1: Take 2mL of carboxylated latex microspheres (150nm, 5% w / v) and add phosphate buffer to 4ml;

[0048] Step 2: Add 50ml 0.2g / L of N-hydroxysuccinimide (NHS) and 50ml 0.2g / L of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride Salt (EDC) for activation, stir well and incubate at 37°C for 20min;

[0049] Step 3: Add 3 mL...

Embodiment 3

[0053] Embodiment three: the accuracy analysis of reagent of the present invention and method:

[0054] Test instrument: Hitachi 7170 automatic biochemical analyzer;

[0055] Test samples: 40 random serum samples and one PGI serum sample (target value 59.75ng / mL)

[0056] Control kit: Protease I (PGI) detection kit (latex immunoturbidimetric method) from a manufacturer approved by the State Food and Drug Administration (including reagents R1 and R2, but the composition is different from the present invention, hereinafter referred to as the control Reagent);

[0057] Using the reagents of Examples 1 and 2 and the comparative reagents, respectively, by their respective detection methods, 40 serum samples and a PGI serum sample (target value of 59.75 ng / mL) were simultaneously measured, and the results are shown in Table 1 and Table 2 Shown:

[0058] Table 1 The results of different detection reagents measuring 40 cases of random serum samples (unit: ng / mL)

[0059]

[006...

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Abstract

The invention discloses a pepsinogen I (PGI) detection kit and a detection method thereof. The detection kit is prepared from a reagent R1 and a reagent R2, wherein the reagent 1 is prepared from 100mmoL / L of phosphate buffer solution, 0.5g / L of Proclinc 300, 10g / L of polyethylene glycol 6000, 1g / L of triton 100 and 10g / L of TRIS; the reagent R2 is prepared from 100mmoL / L of phosphate buffer solution, 1.3mg / mL of latex particle anti-human PGI antibody fixation emulsion (anti-human pepsinogen I (mouse) monoclonal antibody sensitive emulsus solution, 0.5g / L of Proclinc 300, 10g / L of bovine serumalbumin (BSA) and 90g / L of glycerium. The detection method comprises the following steps: separating a serum sample, adding the reagent R1 into the serum sample, evenly mixing, incubating for 3 to 5min, adding the R2, incubating 30s, detecting an absorbance value A1 under 700nm wavelength, detecting an absorbance value A2 under the 700nm wavelength after 5min and calculating a content of the pepsinogen I according to serum standard substance data. The detection kit disclosed by the invention obviously improves flexibility and stability of testing reagents and has the characteristics of high flexibility, high accuracy, stable reagents and the like.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a pepsinogen I (PGI) detection kit and a detection method thereof. Background technique [0002] Pepsinogen (Pepsinogen, PG) is the inactive precursor of pepsin in gastric juice. It is a single-chain polypeptide with a molecular weight of 42000 Da, which is converted into active pepsin in the stomach. According to biochemical properties, immunogenicity, cell origin and distribution in tissues, pepsinogen can be divided into two subgroups, pepsinogen I (PGI) and pepsinogen II (PGII). , known as PGI (PGA), mainly secreted by the principal cells of the fundic glands and cervical mucus cells; 6-7 components with similar immunogenicity, called PG II (PGC), in addition to being secreted by the above two cells, pyloric The mucous cells of the gland, the cardiac gland and the Brunner gland in the upper part of the duodenum can also produce PGII, and the PGII synthesized by the gastr...

Claims

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Application Information

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IPC IPC(8): G01N33/573G01N33/68G01N33/577G01N21/31
CPCG01N33/573G01N21/31G01N33/577G01N33/68G01N2333/96477
Inventor 刘光华杨玉军刘秋明许翠
Owner 广州市伊川生物科技有限公司
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