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Fluorescent immunochromatographic reagent kit for detecting human helicobacter pylori antibody and preparation method of reagent kit

A technology of Helicobacter pylori and fluorescence immunochromatography, applied in biological testing, fluorescence/phosphorescence, material analysis by optical means, etc., can solve the problems of long time, complicated preparation method, high price, etc. Simple, fast and low-cost effects

Inactive Publication Date: 2019-02-15
BIOHIT BIOTECH HEFEI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Existing detection kits for Helicobacter pylori have problems such as complex preparation methods, long time-consuming, and expensive

Method used

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  • Fluorescent immunochromatographic reagent kit for detecting human helicobacter pylori antibody and preparation method of reagent kit
  • Fluorescent immunochromatographic reagent kit for detecting human helicobacter pylori antibody and preparation method of reagent kit
  • Fluorescent immunochromatographic reagent kit for detecting human helicobacter pylori antibody and preparation method of reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] The present embodiment Helicobacter pylori detection kit, such as figure 1 As shown, it includes a support 1, an imprinted membrane 2 arranged on the support 1, an absorbent pad 3 arranged at one end of the imprinted membrane 2, a binding pad 4 at the corresponding other end, and a sample pad 5 arranged at the other end of the binding pad 4, Set on the other end of the sample pad 5 is a blood filter membrane 6 . Wherein, the support 1 is made of a plastic plate, and the imprinted membrane 2 is made of a nitrocellulose membrane. A detection line 21 and a control line 22 are set on the nitrocellulose membrane, the detection line 21 is coated with Caga A, urease and vacA recombinant antigen mixture; the control line 22 is coated with goat anti-mouse IgG. The binding pad 4 is sprayed with fluorescent latex microspheres coupled with mouse anti-human IgG. The mixed mass ratio of Caga A, urease and vacA recombinant antigen coated on the nitrocellulose membrane detection line...

Embodiment 2

[0072] The difference between the Helicobacter pylori detection kit in this example and Example 1 is that during the process of coupling the microspheres to the antibody, the microspheres were coupled on a shaking table at 100-150 rpm at 37°C for 1 hour, and 500uL was directly added for blocking after coupling without centrifugation. The active groups of the microspheres were blocked with liquid, and blocked on a shaker at 150 rpm at 37°C for 30 min. Others are the same as embodiment 1. Embodiment 1 and embodiment 2 detect the same sample, and the data are shown in Table 1.

[0073] Table 1

[0074]

[0075]

[0076] The coefficient of variation CV of the detected concentration is less than 15% when the same sample is detected in the embodiment 1 and the embodiment 2, and there is no significant difference in the detection results of the two reagent strips in the embodiment 1 and the embodiment 2.

Embodiment 3

[0078] The Helicobacter pylori antibody was diluted into 6 concentration gradients of 1.56ug / L, 3.13ug / L, 6.25ug / L, 12.5ug / L, 25ug / L, and 50ug / L as the standard solution, and a blank solution was prepared at the same time.

[0079] Take 80 microliters each of 6 standard products and blank solutions and add them to the sample area (sample pad) of the reagent strip, and insert them into the HitFia-Biohan Fluorescence Immunoassay Analyzer after 15 minutes (Biohan Biotechnology (Hefei) Co., Ltd. ) readings, each standard was detected 3 times. Read the luminescence value and T / C value of the detection line (T line) and control line (C line), see Table 2. Calculate the relationship between the T / C value and the concentration of the corresponding standard substance, draw the standard curve, see figure 2 . The standard curve regression equation is shown in Table 3. The detection limit can reach 0.5ug / L.

[0080] Table 2

[0081]

[0082] Table 3 Four-parameter Logistic curve ...

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Abstract

The invention belongs to the technical field of immunochromatographic diagnosis of in vitro diagnosis reagents, and particularly relates to a fluorescent immunochromatographic reagent kit for detecting a human helicobacter pylori antibody and a preparation method of the reagent kit. The reagent kit comprises a support, a blotting membrane fixed on the support, a water absorption pad fixed at one end of the blotting membrane, and a combining pad fixed at the other end of the blotting membrane, wherein a sample pad is fixed at the other end of the combining pad; a blood filter membrane is fixedat the other end of the sample pad; a detection line coated with a Caga A, urease and cell vacuolating cytotoxin recombinant antigen mixture and a control line coated with sheep anti-mouse IgG (immunoglobulin G) are arranged on the blotting membrane; and a mouse anti-human IgG fluorescent emulsion microsphere is sprayed onto the combining pad. The reagent kit achieves quick and sensitive quantitative detection of the helicobacter pylori antibody by a fluorescent immunochromatographic method through an indirect method principle and fluorescent characteristics of the fluorescent emulsion microsphere.

Description

technical field [0001] The invention belongs to the technical field of immunochromatographic diagnosis of in vitro diagnostic reagents, and in particular relates to a fluorescent immunochromatographic kit for detecting human Helicobacter pylori antibody and a preparation method thereof. Background technique [0002] Helicobacter pylori (Helicobacter pylori, HP) is a Gram-negative microaerophilic bacterium that parasitizes the stomach and duodenum. Its infection is very common, and the infection rate of the natural population in the world exceeds 50%. Almost all Helicobacter pylori-infected patients have chronic active inflammatory reactions histologically. The latest expert consensus defines HP gastritis as "an infectious (infectious) disease", but more than 70% of them have no symptoms. Symptoms, others can lead to chronic gastritis, peptic ulcer, etc. Common symptoms include upper stomach discomfort and pain, flatulence, anorexia, nausea, vomiting, and dark or tar-colored ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/558G01N33/533G01N21/64
CPCG01N21/6486G01N33/533G01N33/558G01N33/6854
Inventor 章西鲁江徽谢龙生刘峰
Owner BIOHIT BIOTECH HEFEI
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