Fluorescence immunochromatography detection test paper strip of trichinellosis as well as preparation method and application thereof
A technique for fluorescence immunochromatography and detection test paper, which is applied in fluorescence/phosphorescence, material analysis by optical means, measurement devices, etc. Effect
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Embodiment 1
[0030] Embodiment 1. Fluorescent immunochromatographic detection test strip for trichinosis.
[0031] A fluorescent immunochromatographic detection test strip for trichinellosis described in this embodiment includes a sample pad, a binding pad, a chromatographic membrane, a water-absorbing pad and a bottom plate, and the binding pad is marked with time-resolved fluorescent microspheres coupled with goat antibody. Pig IgG; the sample pad is provided with a sample hole; the chromatographic membrane is provided with a detection line and a quality control line, wherein the detection line is sprayed with Trichinella spiralis muscle larva ES mixed antigen, and the Trichinella spiralis muscle larvae ES mixed antigen The antigen is composed of the ML-ESP antigen of the excreted secretion of the muscle larvae of Trichinella spiralis and the iML-ESP antigen of the excreted secretion of the muscle larvae of the intestinal stage of Trichinella spiralis; the quality control line is sprayed ...
Embodiment 2
[0032] Example 2. The preparation method of the fluorescent immunochromatographic detection test strip for trichinellosis.
[0033] 1) Preparation of ES mixed antigen from Trichinella spiralis muscle larvae: ML-ESP antigen excreted by Trichinella spiralis muscle larvae and iML-ESP antigen excreted by Trichinella spiralis intestinal stage muscle larvae were mixed according to the mass ratio of 1:1 to obtain Trichinella spiralis Caterpillar muscle larva ES antigen; the specific method is:
[0034] Trichinella spiralis muscle larvae excretion secretion (ML-ESP) antigen preparation:
[0035] The collected Trichinella spiralis muscle larvae were washed three times with a culture solution containing penicillin (500U / mL) and streptomycin (500U / mL) (natural precipitation, each time for 20min), and then placed in a culture medium containing penicillin (250U / mL) and streptomycin (250U / mL) in culture medium (density 5000 / mL), 37°C, 10% carbon dioxide, cultured for 18h-24h and then colle...
Embodiment 3
[0073] Example 3. The application of trichinellosis fluorescent immunochromatographic detection test strip in the detection of trichinellosis.
[0074] Sample dilution: Dilute Trichinella antibody-positive serum (such as pig serum infected with Trichinella), negative serum (such as normal pig serum) and serum to be tested (such as pig serum) samples with sample diluent, respectively, to 95 μL diluent Add 5 μL of porcine serum and mix with a pipette. Place the test strip flat on the operating table with the sample well facing up, add 100 μL of diluted serum sample into the well, and let it stand at room temperature for 5-10 minutes. The sample diluent is prepared by adding 9 g of sodium chloride (NaCl), 2 g of bovine serum albumin (BSA), and adding double distilled water to 1000 mL. Use UV lamp 360-370nm to irradiate the test strip to observe the results.
[0075] The results should be judged within 20 minutes after adding the sample. If the quality control line and detectio...
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