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Fluorescent silica labeled immunochromatographic test paper for quantitative gentamicin detection and preparation method

An immunochromatographic test paper and silica technology, which is applied in measurement devices, analytical materials, instruments, etc., can solve the problems of being unsuitable for large-scale sample screening, long time required for determination, low sensitivity, etc., and achieve low detection costs. , Easy to operate, to overcome the effect of low sensitivity

Active Publication Date: 2015-07-01
HENAN ACAD OF AGRI SCI
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  • Description
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Problems solved by technology

Although the microbiological method is simple in principle and easy to operate, the time required for the determination is too long and the sensitivity is not high
The physical and chemical methods for detecting gentamicin residues mainly include high performance liquid chromatography (HPLC), mass spectrometry (MS) and gas chromatography. Although these methods are sensitive and accurate, they are expensive, time-consuming, and require high technical Reasons not suitable for screening of large batches of samples

Method used

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  • Fluorescent silica labeled immunochromatographic test paper for quantitative gentamicin detection and preparation method
  • Fluorescent silica labeled immunochromatographic test paper for quantitative gentamicin detection and preparation method
  • Fluorescent silica labeled immunochromatographic test paper for quantitative gentamicin detection and preparation method

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preparation example Construction

[0041] (2) Preparation of fluorescent antibody fiber layer

[0042] a. Preparation of fluorescent silica nanoparticles

[0043] 47mmol aminopropyltriethoxysilane (APTS), 7.1 mmol BHHCT (4,4-bis(1", 1",1",2",2",3",3"-heptafluoro-4", 6"-hexanedion-6"-yl)-chlorosulfo-o-terphenyl, molecular formula C 30 h 15 CIF 14 o 6 S, molecular weight 804.93) and 3.55mmol EuCl 3 ?6H 2 O was added to 30 μL cyclohexane, and after ultrasonic oscillation for 15 minutes, the reaction solution was added to the W / O microemulsion containing 1.1 mL water, 4.74 g Triton X-100, 3.64 g n-octanol and 14.50 g cyclohexane , after stirring for 0.5 hours, add 200 μL of TEOS (tetraethoxysilane) and 200 μL of concentrated ammonia water, stir at room temperature for 24 hours, then add 40 mL of acetone to end the reaction. The supernatant was removed after the reaction liquid was centrifuged, and the precipitated part was washed three times with ethanol and double distilled water respectively, and then susp...

Embodiment 1

[0060] Embodiment one: see figure 2 , image 3. In the figure, the support layer 1 is made of plastic sheet strips, the adsorption fiber layer 2 is made of glass fiber cotton, the fluorescent antibody fiber layer 3 is adsorbed with fluorescent antibody glass fiber cotton of anti-GM monoclonal antibody, and the cellulose membrane layer 4 is made of nitric acid The cellulose membrane, the water-absorbing material layer 5 at the handle end is made of water-absorbing filter paper, and the layers of the absorbent fiber layer 2, the fluorescent antibody fiber layer 3, the cellulose film layer 4, and the water-absorbing material layer 5 are pasted and fixed on the support layer 1 from left to right On the other hand, the fibers at the junctions between the layers interpenetrate each other. On the cellulose membrane layer 4, there is a stealth detection blot 6, which is made with GM-coupled bovine serum albumin solution (BSA); the stealth control blot 7 is blotted on the cellulose ...

Embodiment 2

[0064] Embodiment 2: The structure of the test paper is basically the same as that of Embodiment 1. The difference is that the fluorescent antibody fiber layer is adsorbed with anti-GM polyclonal antibody, the adsorption fiber layer is made of nylon membrane, and the cellulose membrane layer is made of pure cellulose membrane. Both the invisible detection imprint and the invisible control imprint are "ten", and the protective film at the handle end covered on the water-absorbing material layer is blue.

[0065] For the detection of milk samples: Dilute the milk samples with physiological saline to make a 1:2-1:5 suspension of the sample to be tested.

[0066] Operation method: insert the sample end of the GM test paper into the sample to be tested, the insertion depth does not exceed the marking line, take out the test paper in about 10 seconds, put it into the fluorescent strip reader after 5 minutes and read the result directly.

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Abstract

The invention relates to a gentamicin immunochromatography test paper marked with fluorescent silica nanoparticles and a preparation method thereof. The bottom layer of the immunochromatography test paper is a supporting layer, the middle layer is an adsorption layer, and the protective layer is fixed on the adsorption layer. The layers from the test end are the adsorption fiber layer, the fluorescent antibody fiber layer, the cellulose film layer and the water-absorbing material layer at the handle end, and the invisible detection blot printed with the carrier protein solution coupled with gentamicin is arranged on the cellulose film layer , and a stealth control blot printed with goat anti-mouse IgG, rabbit anti-mouse IgG or goat anti-rabbit IgG antibody solution or Staphylococcus aureus protein A solution; the fluorescent antibody fiber layer is made of glass fiber cotton that absorbs fluorescent antibodies , the fluorescent antibody was made using gentamicin antibody labeled with fluorescent silica nanoparticles. The immunochromatographic test paper of the invention is simple and fast, realizes sensitive and fast on-site detection of gentamicin, has wide application range, low detection cost, and is easy to popularize and apply.

Description

technical field [0001] The invention relates to an immunochromatographic test paper, in particular to an immunochromatographic test paper for quantitatively detecting gentamicin based on fluorescent silicon dioxide nanoparticles and a preparation method thereof. Background technique [0002] Gentamicin (Gentamicin, GM) is a kind of aminoglycoside antibiotics, which mainly acts on Gram-negative bacteria. Because of its broad antibacterial spectrum, good curative effect and low price, it is widely used in veterinary clinics. Endometritis has a good curative effect, and it is also used as a feed drug additive in compound feed. Irregular drug use will cause drug residues in animal products, especially milk of lactating animals, which poses potential hazards to human health, causing ototoxicity and kidney toxicity. The maximum residue limit of gentamicin in animal food has been stipulated successively at home and abroad. According to the "Maximum Residue of Veterinary Drugs in A...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/533
Inventor 张改平王方雨宋春美职爱民胡骁飞赵东
Owner HENAN ACAD OF AGRI SCI
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