A rapid detection test strip of Fasciola large fasciola immunochromatography and its preparation method
A technology for detecting test strips and immunochromatography, which is applied in the directions of measuring devices, analytical materials, instruments, etc., can solve the problems of high price and complicated operation, and achieve the effects of simple operation, good effect and wide application range.
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[0031] B. Preparation of rabbit anti-mouse immunoglobulin antibody: Mix mouse immunoglobulin with a mass concentration of 1 mg / mL with Freund's complete adjuvant 1:1, and subcutaneously apply 1 ml / rabbit to the paw of the rabbit. Injection immunization; 7 days later, the mouse IgG with a mass concentration of 2 mg / ml was mixed with incomplete Freund's adjuvant 1:1, and the above-mentioned rabbits were injected with a secondary immunization in the popliteal lymph node at a dose of 0.5 mL / rabbit; 10 Days later, the immunization was boosted once by the method of secondary immunization. After immunization, blood was collected from ear vein regularly to measure the serum titer. After the serum titer was reached, blood was collected from the heart, and the serum was separated; after preliminary purification by the conventional octanoic acid-saturated ammonium sulfate method, Then use the portion G protein column for further purification; when the titer of rabbit anti-mouse immunoglob...
Embodiment 1
[0046] The colloidal gold-labeled 7D1 monoclonal antibody against Fasciola maxima ES antigen prepared by the above method was diluted 1:5, and sprayed on the labeling pad in an amount of 12.5 μL per pad for coating, and then the labeling pad was baked at 37°C. Store dry and sealed.
[0047] The 7D4 monoclonal antibody against the ES antigen of Fasciola maxima was diluted to a concentration of 0.26 mg / mL, and the amount of 1.5 μL was coated on the coated detection line T; the rabbit anti-mouse immunoglobulin antibody was diluted to 0.30 mg / mL The concentration of 1.5μL was coated to the quality control line C of the coated film, dried at 37°C, and sealed for storage.
[0048] The sample pad 2, the marking pad 3, the coating film 4, and the water-absorbing pad 5 are lapped and pasted on the support liner made of PVC material in sequence, and the fasciola immunochromatographic rapid detection test strip is completed.
[0049] The immunochromatographic rapid detection test strip ...
Embodiment 2
[0051] Validation assay for detection sensitivity to ES antigens of Fasciola grandis. The test strip was prepared according to the preparation method of the rapid detection test strip for the detection of Fasciola maxima by immunochromatography described in Example 1. After purification of Fasciola maxima ES antigen, the antigen concentration was determined by UV spectrophotometer to be 0.94 gm / mL. : 12800 was double-diluted and detected with test strips, each dilution was detected with three test strips, and a parallel control was set up, the results are shown in Table 1.
[0052] Table 1 Test results of test strip detection sensitivity
[0053] Antigen dilution 200 400 800 1600 3200 6400 12800
[0054] Antigen concentration (μg) 47.00 2.35 1.18 0.59 0.29 ¬¬— —
[0055] T-line Color development Color development Color development Color development No color development No color development
[0056] C-line color rendering color rendering color rendering color rendering colo...
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