Immunofluorescence chromatography detection card and method for simultaneous detection of florfenicol and trimethoprim in poultry eggs
A technology of florfenicol and trimethoprim, which is applied in the field of biological detection, can solve the problems of only one of them being detected, high requirements for samples to be tested, and threats to public health, so as to improve sensitivity and labeling efficiency , Reduce the effect of steric hindrance effect
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Embodiment 1
[0066] In this embodiment, the immunofluorescent chromatography detecting card in which the fluorophenicon, the methoxymethridin content is synchronized, including the bottom plate, and the bottom plate, and the bottom plate is connected to the left to right, and the water absorbing paper pad, the reaction film, the combined pad, The sample pad, the reaction film was prepared with fluoropheniconi detection line T1, the methoxybenzil detection line T2, and the heat row of three lines, the binding pad was covered with fluorine tissue detection microspheres. Methylene anti-chicken IGY polyclonal antibody labeling microspheres;
[0067] Fluornisi detection microspheres are long-chain biotinoflu fluoropheniconi monoclonal antibodies and streptavidin fluorescent microspheres with streptavidin fluorescent microspheres by connecting polyethylene glycol long-chain excess arms (-DPEG24-). Streptaviarin is combined;
[0068] Methoxybenzeridine detection microspheres are ultra-chain biotin-li...
Embodiment 2
[0106] This embodiment provides a method of detecting a fluorine-based, methoxyenin content synchronously detecting a fluorine content in an egg, and specifically comprises the steps of:
[0107] (1) Pre-treatment of the sample first, the specific steps are: Take 2G to detect homogeneous eggs that do not contain fluorine-containing and methoxybenzil as a negative reference, 3 ml of acetic acid per tube. Ester (containing 1 wt% trichloroacetic acid), manual shock 30s, 4000 rpm, centrifugation for 1 min. Take 1 mL of the supernatant to 15 ml of centrifuge tube, 5 ml of n-hexane, 0.5 ml of reconstitution, and the lower layer water phase to be tested;
Embodiment 3
[0110] Preparation of the standard
[0111] Series standard preparation: Take 16 μL of fluorine tissue standard (10 mg / kg) and 16 μL of methoxymethridine standard (10 mg / kg) to add to 968 μl of methanol solution, mixed with mixed standard 1 (fluorbennette It is 160 μg / kg, methoxybazine is 160 μg / kg), and then doubles 5 times to mix the standard 2, mixed standard 3, mixed standard 4, mixed standard 5, mixed standard 6; take 7 15 ml of centrifuge tube, 1 ml of white samples without fluorine-free and methoxy, and add 50 μl of mixed standard 1-6 to 6 tubes, and 50 μl of methanol is added to mix.
[0112] The fluvovedronidin concentration gradient is: 4 μg / kg, 2 μg / kg, 1 μg / kg, 0.5 μg / kg, 0.25 μg / kg, 0.125 μg / kg, 0 μg / kg;
[0113] The formation of the methoxybenzil concentration is: 4 μg / kg, 2 μg / kg, 1 μg / kg, 0.5 μg / kg, 0.25 μg / kg, 0.125 μg / kg, 0 μg / kg;
[0114] The 3 ml extract was added, and the hand was shaken for 30s, 4000 rpm, centrifuged for 2 ...
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