Fluorescent silica labeled immunochromatographic test paper for quantitative gentamicin detection and preparation method
A technology of immunochromatographic test paper and silicon dioxide, which is applied in the direction of measuring devices, analytical materials, instruments, etc., can solve the problems of not being suitable for large-scale sample screening, long time required for measurement, high technical requirements, etc., and achieve low detection cost , Easy to operate, overcome the effect of low sensitivity
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[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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