Pepsinogen I and II combined detection method and kit thereof

A combined detection technology of pepsinogen, which is applied in the field of combined detection methods and kits of pepsinogen Ⅰ and Ⅱ, can solve the problems of undetectable, high reagent cost, and long time consumption, and achieve the effect of reducing the impact of impurities

Inactive Publication Date: 2015-04-08
江苏宏泰格尔生物医学工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The ELISA method is accurate in detection, can be accurately quantified, and has high sensitivity. However, the detection of samples requires steps such as sample addition, warm bath, washing, color development, and termination. The operating procedures are cumbersome and time-consuming. It takes at least 40 minutes, not suitable for large-scale physical examination screening; sample detection requires a plate washer and microplate reader, which is inconvenient to carry, so it cannot be detected in the patient's home or in an ambulance; in addition, its sensitivity and specificity are not high enough for low concentration The accuracy of sample detection is not high
However, chemiluminescence immunoassay, although it has high accuracy and fast detection speed, requires expensive chemiluminescence immunoassay analyzers and specific analysis rooms. In addition, the cost of reagents is relatively high, so it cannot be widely used in lower-level hospitals and medical institutions. In the institution

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  • Pepsinogen I and II combined detection method and kit thereof
  • Pepsinogen I and II combined detection method and kit thereof
  • Pepsinogen I and II combined detection method and kit thereof

Examples

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Effect test

Embodiment 1

[0035] The dual-wavelength fluorescent immunochromatographic combined detection method of pepsinogen I and II of the present invention comprises the following steps:

[0036] 1) Preparation of immunochromatographic test strips: Coat pepsinogen Ⅰ monoclonal antibody, pepsinogen Ⅱ monoclonal antibody and chicken IgY on detection line 1 (T line 1 ), detection line 2 (T line 2) and control line (C line), after drying for 1 to 5 hours, cut with a strip cutter to obtain immunochromatographic test strips with a width of 3 to 5 mm;

[0037] It should be noted that: a) Traditional qualitative immunochromatography uses coated goat anti-mouse antibody as the control line. With the increase of pepsinogen Ⅰ or some anti-mouse antibody blocking agents in the serum, the signal on the control line As it decreases, its signal value cannot be used for calculation, and the accuracy of the signal of the test line has no reference basis. The present invention adopts chicken IgY antibody and goat ...

Embodiment 2

[0048] like Figures 1 to 2As shown, the dual-wavelength fluorescent immunochromatographic detection kit of pepsinogen I of the present invention includes a kit body, a cryopreservation tube, and a diluent bottle, and the kit body includes a plastic liner 1 and is fixed on the plastic liner. The sample pad 2, the immunochromatography test strip 3 and the absorbent paper 6, the immunochromatography test strip is made of nitrocellulose membrane material, the sample pad and the absorbent paper are respectively lapped on both sides of the immunochromatography test strip, and the immunochromatography test strip is The chromatography test strip is provided with detection line 1 (T1) 4, detection line 2 (T2) 11 and control line 5, and the detection line 1, detection line 2 and control line are respectively coated with pepsinogen Ⅰ monoclonal antibody , pepsinogen Ⅱ monoclonal antibody and chicken IgY; freeze-dried probes were stored in the cryopreservation tube, and the lyophilized p...

specific Embodiment

[0052] The present invention also provides a specific embodiment of dual-wavelength fluorescence immunochromatographic detection of pepsinogen I, which includes the following steps in sequence:

[0053] Step 1: Preparation of lyophilized probes

[0054] 1) Mix two fluorescent latex particle solutions with emission wavelengths of 550nm and 700nm respectively according to the volume ratio of 1:1. After mixing evenly, take 500 μl of mixed fluorescent latex particle solution (containing carboxyl groups) and use pH6.0 MES buffer After washing and centrifuging for three times, the precipitate was diluted with pH6.0 MES buffer, mixed with 10 mg EDC, and then activated at room temperature for 30 minutes. After centrifugation, the precipitate was washed three times with pH6.0 MES buffer, and then the precipitate was washed with pH6. Dilute with 0MES buffer, add 125 μg pepsinogen Ⅰ monoclonal antibody, react at room temperature for 3 hours, add BSA to block, continue to react for 30 min...

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Abstract

The invention relates to a pepsinogen I and II combined detection method and a kit, and especially relates to a dual wavelength fluorescence immunity chromatography combined detection method of pepsinogen I and II and a detection kit thereof. The method comprises the following steps: 1)preparing an immunity chromatography test strip; 2)preparing a freeze-dried probe; 3)preparing a sample weak solution; and 4)examining the sample. The pepsinogen I and II combined detection method and the kit have the advantages of high sensitivity, high accuracy, simple operation and low cost.

Description

technical field [0001] The invention relates to a combined detection method and kit for pepsinogen I and II, in particular to a dual-wavelength fluorescent immunochromatographic combined detection method for pepsinogen I and II and a detection kit thereof. Background technique [0002] Pepsinogen (PG), the precursor of pepsin, is a single-chain polymorphism with a molecular weight of 42,000 Da. It is divided into two subgroups according to its biochemical properties and immunogenicity. Components 1-5 have the same immunogenicity and are called PGⅠ, which are mainly secreted by the principal cells and mucus neck cells of the gastric gland, and most of them enter the gastric cavity; components 6-7 are called PGⅡ, which are produced by the oxyntic glands of the gastric fundus The chief cells of the oxyntic glands, the mucous neck cells of the oxyntic glands, the mucous cells of the cardia glands and the pyloric glands of the gastric antrum, and the Brunner glands of the upper d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573G01N33/533G01N21/64
CPCG01N33/558G01N21/6486G01N33/531G01N33/68
Inventor 廖平璋张翼飞张鹏张华
Owner 江苏宏泰格尔生物医学工程有限公司
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