Fluorescent microsphere immunity chromatography test paper for detecting food-borne pathogenic microbe

A technology for immunochromatographic test strips and pathogenic microorganisms, which is applied in measurement devices, analytical materials, instruments, etc., can solve the problems of low contrast between color bands and background, difficulty in realizing multiple inspections and joint inspections, and inability to realize quantitative detection, etc. Achieve rapid and accurate qualitative and quantitative detection, rapid detection, and low prices

Inactive Publication Date: 2009-06-24
WUXI ZODOLABS BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] (1) The colloidal gold labeling process is an electrostatic adsorption process, which is a physical adsorption, so the stability in the liquid phase is poor, often causing the labeled protein molecules to fall off again
[0007] (2) The test result is judged by displaying a single purple-red strip, and the color is single, so it is difficult to realize multiple inspection and joint inspection
[0008] (3) Only when the gold particles gather to a certain amount, the human naked eye can observe the purple-red band, ...

Method used

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  • Fluorescent microsphere immunity chromatography test paper for detecting food-borne pathogenic microbe
  • Fluorescent microsphere immunity chromatography test paper for detecting food-borne pathogenic microbe
  • Fluorescent microsphere immunity chromatography test paper for detecting food-borne pathogenic microbe

Examples

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Effect test

Embodiment 1

[0064] Example 1: Preparation of Fluorescent Microsphere Immunochromatographic Test Strips for Detecting Listeria monocytogenes in Beef Stuffing Samples in Farmer's Vegetable Market

[0065] 1. Preparation of chromatography test strips

[0066] 1. Preparation of nitrocellulose membrane;

[0067] Preparation of detection area and quality control area:

[0068] Monocytogenes polyclonal antibody and anti-mouse antibody were coated onto nitrocellulose membrane: with 0.1M PBS pH7.2 (phosphate buffered saline containing 5% sucrose and 0.05% Tween-20 ( Tween-20)) adjusts the concentration of Listeria monocytogenes polyclonal antibody to be 0.5mg / mL, and the resulting solution is sprayed on the membrane as the detection area, and the PBS of 0.01MpH7.2 (which contains 5% sucrose and 0.05 % Tween-20 (Tween-20)) adjusts the concentration of anti-mouse antibody to be 0.5mg / mL, and the solution of gained is sprayed on the film as the quality control area, and the amount of spray film in ...

Embodiment 2

[0079] Embodiment two: the preparation of the fluorescent microsphere immunochromatography test strip of detecting Escherichia coli O157:H7 in the milk sample;

[0080] 1. Preparation of chromatography test strips;

[0081] 1. Preparation of nitrocellulose membrane;

[0082] Preparation of detection area and quality control area: adjust the concentration of Escherichia coli O157:H7 polyclonal antibody to 1.0 mg / mL with 0.05M PBS pH7.2 (containing 5% sucrose and 0.05% Tween-20), and the resulting solution Spray on the membrane as the detection area, adjust the concentration of anti-mouse antibody to 1.0 mg / mL with 0.01M PBS pH7.2 (which contains 5% sucrose and 0.05% Tween-20), and spray the resulting solution on the membrane as the detection area. In the control area, the amount of spray film in the two areas is 0.74μL / cm, the two areas are separated by 5mm, and the quality control area is 2mm away from the end of the nitrocellulose membrane. After drying at 37°C overnight, st...

Embodiment 3

[0093] Example 3: Preparation of fluorescent microsphere immunochromatographic test strips for detecting Staphylococcus aureus and Listeria monocytogenes in pork samples

[0094] 1. Preparation of chromatography test strips

[0095] 1. Preparation of nitrocellulose membrane;

[0096] Detection area preparation: adjust the concentration of Listeria monocytogenes monoclonal antibody to 0.5mg / mL with 0.01M PBS of pH7.2, and adjust the concentration of Staphylococcus aureus monoclonal antibody to 0.8mg / mL, and then the obtained The solution was sprayed on the membrane to form two detection lines, the amount of the sprayed membrane was 0.74 μL / cm, the distance between the two lines was 5mm, and the distance between the absorbent paper was 2mm, after drying at 37°C overnight, it was stored in a dry environment at room temperature for later use.

[0097] 2. Preparation of fluorescent microsphere pads;

[0098] Preparation of the polyclonal antibody against Listeria monocytogenes la...

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Abstract

The invention discloses a fluorescent microsphere immunity-chromatography test strip for detecting food-borne pathogenic microorganisms in a rapid, specific, qualitative and quantitative manner and a preparation method thereof. Filter paper, a sample pad, a glass fiber membrane spray-painted by fluorescent microsphere marked food-borne pathogenic microorganism monoclonal antibodies or polyclonal antibodies, a nitrocellulose membrane and drinking paper are stuck to an underplate orderly in a related joint manner; the food-borne pathogenic microorganism monoclonal antibodies or the polyclonal antibodies are coated on the nitrocellulose membrane and are used as a detection area; and anti-mouse antibodies are coated on the nitrocellulose membrane and used as a quality control area. The test strip has the preparation steps as follows: (1) samples to be detected are processed; (2) the nitrocellulose membrane is prepared; (3) a fluorescent microsphere pad is prepared; (4) the test strip is assembled; and during the detection process, the CCD scanning technique is utilized to collect all emission spectrums, and then the emission spectrums are analyzed through a fluorescence analyzer and relevant software after accumulation and multiplication, so that the quantitative detection is realized. The fluorescent microsphere immunity-chromatography test strip and the preparation method thereof are mainly used for qualitative and quantitative detection of all the food-borne pathogenic microorganisms in food safety inspection.

Description

technical field [0001] The invention belongs to the field of detection of food-borne pathogenic microorganisms in food safety, and specifically relates to a fluorescent microsphere immunochromatographic test strip for qualitatively and quantitatively detecting food-borne pathogenic microorganisms in a sample and a preparation method thereof. Background technique [0002] In today's era of increasingly frequent global population movement and increasingly strong food trade, foodborne microbial poisoning, as an important factor threatening human health, has increasingly attracted the attention of scientific and technological circles around the world. A large number of statistical data and literature data show that Escherichia coli O157:H7, Salmonella, Listeria monocytogenes, Staphylococcus aureus, Shigella, botulinum, Vibrio parahaemolyticus, Proteus, enterocolitis Microorganisms such as Yersinia, Vibrio cholerae, Streptococcus faecalis, pathogenic Escherichia coli and Cryptosp...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N33/558G01N33/545
Inventor 赖卫华熊勇华魏华金晶
Owner WUXI ZODOLABS BIOTECH
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