Escherichia coli O157:H7 direct type immunofluorescence chromatography test paper
A kind of technology of O157, Escherichia coli
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Embodiment 1
[0036]The preparation process of the test strip of the present invention comprises the steps of: preparation of Escherichia coli O157:H7 monoclonal or polyclonal antibody, preparation of the adsorption fiber layer, preparation of the cellulose film layer, assembly of the test strip and the like.
[0037] (1) Preparation of anti-Escherichia coli O157:H7 monoclonal antibody or polyclonal antibody
[0038] Monoclonal antibody preparation: at an inactivating concentration of 10 8 Escherichia coli O157:H7 cfu / mL whole bacteria immunized 6-8 week-old Balb / C mice 3-4 times, with an interval of 3-5 weeks between each immunization. After confirming that the antibody titer met the requirements, super-strong immunization was performed, and then 3 On ~4 days, blood was collected from the infraorbital sinus of the immunized mice, and the positive serum was separated; the mice were killed by dislocation of the neck, and the mice were soaked in 75% alcohol for 5-10 minutes to disinfect the b...
Embodiment 2
[0051] Example 2: see image 3 , Figure 4 . In the figure, 1 is the support layer, which is made of plastic sheet strips, 2 is the absorbent fiber layer, which is made of glass fiber cotton, the cellulose membrane layer 3 is made of nitrocellulose membrane, and the water-absorbing material layer 4 at the handle end is made of water-absorbing filter paper 1. Paste and fix the absorbent fiber layer 2, the cellulose membrane layer 3, and the water-absorbing material layer 4 on the support layer 1 from left to right, and the fibers at the junctions between the layers interpenetrate with each other. On the cellulose film layer 3, there is a stealth detection imprint 5, which is made of Escherichia coli O157:H7 monoclonal antibody; the imprinted band is formed as "︱".
[0052] 6-1 is the white protective film covering the sample end on the absorbent fiber layer 2, 6-2 is the protective film of other colors (such as yellow) covering the water-absorbing material layer 4, 7 is the s...
Embodiment 3
[0060] Embodiment 3: The structure of the test strip is the same as that of Embodiment 2. Preparation and testing steps of samples to be tested:
[0061] Test bread: aseptically take 25g sample and add it to 225mLEC or NB broth containing FTIC solution, put the above solution into a homogenizer, homogenize continuously for 1-2min on a slap-type homogenizer, and incubate at 37°C several hours.
[0062] Operation method: Insert the test paper into the sample solution, take it out after 10-20 seconds, and lay it flat. After 5-10 minutes, observe whether there is fluorescence on the test line of the test paper to determine whether the sample contains E. coli O157:H7, or read the strip by fluorescence instrument for semi-quantitative analysis.
[0063] Result judgment: (a) positive if there is a yellow-green blot "︱" on the cellulose membrane, it means that the test result is positive, indicating that E. coli O157:H7 is contained in the sample to be tested; There is no yellow-gr...
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