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Fluorescent nano-immunochromatography kit for quickly detecting vibrio parahaemolyticus and preparation method

A technology of hemolytic Vibrio and immunochromatographic test paper, which is applied in the direction of fluorescence/phosphorescence, material analysis through optical means, and measurement devices, which can solve the problems of expensive equipment, numerous reagents, and cumbersome processes, and achieve low cost and high detection efficiency. The process is simple and fast, and the effect of high sensitivity

Inactive Publication Date: 2016-03-30
珠海国际旅行卫生保健中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional culture identification method has the characteristics of accurate and reliable results and does not require complicated instruments. However, due to its defects such as long detection period, cumbersome process, and various reagents, it is far from meeting the needs of modern rapid detection.
Various rapid detection methods based on the principles of immunology, biomedicine, and molecular biology methods have the advantage of being fast, but they cannot meet the needs of on-site detection due to defects such as the need for expensive equipment and the need to bring them back to professional laboratories for testing.

Method used

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  • Fluorescent nano-immunochromatography kit for quickly detecting vibrio parahaemolyticus and preparation method
  • Fluorescent nano-immunochromatography kit for quickly detecting vibrio parahaemolyticus and preparation method
  • Fluorescent nano-immunochromatography kit for quickly detecting vibrio parahaemolyticus and preparation method

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preparation example Construction

[0031] The preparation method of the above-mentioned kit comprises the preparation steps of reaction micropore reagents and the preparation steps of immunochromatographic test strips; the preparation steps of said reaction micropore reagents are:

[0032] (1) Modification of magnetic fluorescent nanoparticles: Add appropriate amount of europium ion-labeled magnetic fluorescent nanoparticles into aminosilane ethanol solution, heat to reflux for 12-24 hours, wash the product after heating and reflux with ethanol. Resuspend nanoparticles with a saturated solution of glutaric anhydride in dimethylformamide (DMF), and stir at room temperature for 2 to 4 hours. The nanoparticles were washed with ethanol again, and the nanoparticles were resuspended in double distilled water to prepare a 1 mg / ml suspension for later use, and the obtained time-resolved magnetic fluorescent nanospheres were modified with amino groups on the surface.

[0033](2) Preparation of magnetic fluorescent nano-...

Embodiment 1

[0040] Example 1: Diarrhea, vomit, drinks, milk and other liquid samples to be tested, directly take 100ul of samples (normal saline or PBS buffer can be added in the state of diarrhea and vomit), and add 100ul of sample diluent into the reaction cup detection.

Embodiment 2

[0041] Embodiment 2: Aquatic products, molded feces, meat, vegetables and fruits and other solid objects to be tested, first weigh a certain amount of samples, add physiological saline or PBS buffer, etc., homogenate, and then dilute with an equal volume Add to the cuvette together for detection.

[0042] The reading of the result is observed under the ultraviolet light within 10 minutes after the test strip absorbs the reaction solution in the reaction cup.

[0043] Test results:

[0044] Specificity: detect Staphylococcus epidermidis, Staphylococcus saprophyticus, Listeria monocytogenes, Salmonella, Shigella, Escherichia coli, O1 group Vibrio cholerae, O139 group cultured overnight with the immunochromatographic method established by the present invention Vibrio cholerae, Vibrio parahaemolyticus, Paratyphi A, B, C, Typhimurium, Shigella, and Escherichia coli O157 were all negative.

[0045] Sensitivity: Dilute the Vibrio parahaemolyticus culture solution with normal saline...

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Abstract

The invention aims at providing a fluorescent nano-immunochromatography kit for quickly, sensitively and simply detecting vibrio parahaemolyticus and a preparation method of the kit. In the invention, the separation and enrichment of vibrio parahaemolyticus are realized through specific combination of an immune functionalized time-resolved magnetic fluorescent nanomaterial and the vibrio parahaemolyticus; the captured to-be-detected bacteria are combined with a vibrio parahaemolyticus-specific polyclonal antibody on an immunochromatography test strip; and the detection of the to-be-detected bacteria in the sample is realized by detecting the fluorescence signals on the test strip under an ultraviolet lamp. The kit provided by the invention can be applied to the field of detection on microbial diarrhea-causing pathogenic bacteria.

Description

technical field [0001] The invention relates to the field of detection of microbial diarrhea-causing pathogenic bacteria, in particular to a fluorescent nano-immunochromatography kit and a preparation method for rapidly detecting Vibrio parahaemolyticus. Background technique [0002] Infectious diarrhea is a group of intestinal infectious diseases caused by bacteria, viruses, protozoa and other pathogens, mainly diarrhea. Due to the high incidence and widespread prevalence of the disease, which affects people's health and brings a heavy economic burden to the society, it has become an important public health problem in the world, especially in developing countries. According to WHO estimates, tens of millions of people suffer from the disease every day in the world, with 3 to 5 billion cases of diarrhea every year, and 5 to 10 million deaths due to severe diarrhea, with an average of 25,000 deaths per day. The incidence of diarrheal diseases has always ranked first in intes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531G01N33/533G01N33/553G01N33/569G01N21/64
CPCG01N33/577G01N21/6408G01N33/531G01N33/533G01N33/553G01N33/56916
Inventor 史咏梅何晖汪海波冯子力伍碧梅涂承宁谭华叶立青
Owner 珠海国际旅行卫生保健中心
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