Immunochromatography test strip for detecting cystic echinococcosis and preparation method thereof
A cystic hydatid disease and immunochromatography technology, which is applied to measurement devices, analytical materials, instruments, etc., can solve the problems of high requirements, time-consuming and laborious, and is not suitable for on-site use, and achieves a simple detection method and broad application prospects. , the effect of low cost
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Embodiment 1
[0053] Embodiment 1, the analysis of the response analysis of the serum of the echinococcosis patient to the crude antigen and the analysis of the dominant specific antibody subclass
[0054] 1. Preparation of natural antigen against cystic echinococcosis
[0055] Take the fresh goat liver encapsulation fluid, centrifuge at 3000r / min for 20min, suck the supernatant into the dialysis bag, dialyze with PBS solution at 4°C overnight, centrifuge the dialyzed encapsulation fluid at 6000r / min for 20min, take the supernatant, The precipitate was discarded, and the crude antigen of the cystic fluid was obtained, which was freeze-dried and stored at low temperature for later use.
[0056] 2. Method: ELISA method was used for analysis, and the crude cystic fluid antigen was used as the coated antigen to react with the serum of echinococcosis patients, non-echinococcosis patients and healthy people respectively, and then the goat anti-human IgG1 antibody horseradish peroxide Enzyme conj...
Embodiment 2
[0060] Embodiment 2, the preparation of the immunochromatography test strip that detects cystic echinococcosis
[0061] 1. The preparation of the natural antigen against cystic echinococcosis is the same as in Example 1.
[0062] 2. Preparation of monoclonal antibodies against dominant specific antibody subclasses of hydatid antigens
[0063] BALB / c mice were immunized with human IgG4 antigen according to the conventional method, splenocytes from the immunized mice were fused with SP2 / 0 tumor cells, and the hybridoma cells with high function and high efficiency secreting specific antibodies were cloned three times and then injected into mice to produce ascites. The obtained monoclonal antibody was purified by the conventional saturated ammonium sulfate salting-out method, and the antibody titer was detected by the immunoagar diffusion method. The results showed that the antibody titer was greater than 1:128, indicating that the obtained monoclonal antibody had good specificity...
Embodiment 3
[0081] Embodiment 3, the preparation of the immunochromatographic test strip that detects cystic echinococcosis
[0082] A colloidal gold-labeled antibody was prepared as a detection probe by using a mouse anti-human IgG4 monoclonal antibody purchased from BD Company of the United States (Product No. 555881), and other steps were the same as in Example 2.
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