Colloidal gold immunochromatography test strip for detecting bovine rotavirus, and preparation method and application thereof
An immunochromatographic test paper and the technology of bovine rotavirus, applied in the field of medicine, can solve the problems that are not conducive to large-scale detection, unsuitable for on-site detection, and high requirements, and achieve good sensitivity, fast detection speed, and convenient storage.
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Embodiment 1
[0085] Example 1 Screening and establishment of hybridoma cells secreting monoclonal antibody against bovine rotavirus VP6 protein
[0086] 1 test material
[0087] 1.1 Strains, cells and viruses
[0088] The recombinant strain pProHTa-NCDV-VP6 / Rosetta expressing bovine rotavirus VP6 protein was constructed and preserved by our laboratory (the recombinant strain has been recorded in the following literature: "Establishment of indirect ELISA detection method for bovine rotavirus VP6 protein", Zhang Yiting et al., Chinese Veterinary Science, 2010, 40(10), 1039-1043); BRV NCDV cell-adapted strains were purchased from China Veterinary Drug Control Institute, and MA104 cells were preserved by our laboratory.
[0089] 1.2 Experimental animals
[0090] SPF grade BALB / c mice and healthy female rabbits weighing 2-3 kg were purchased from Liaoning Changsheng Biotechnology Co., Ltd.
[0091] 1.3 Reagents
[0092] (1) Solution I: 12g urea, 1.56g NaH 2 PO 4 2H 2 O and 0.12g Tris-Bas...
Embodiment 2
[0145] Embodiment 2 is used to detect the preparation of the colloidal gold immunochromatography test strip of bovine rotavirus
[0146] 1 material
[0147] 1.1 Viruses and Cells
[0148] Bovine rotavirus NCDV strains, PoRV, TGEV, PEDV, and BPV are all preserved by our laboratory. MA-104 cells and SP2 / 0 cells were preserved by the laboratory; bovine rotavirus VP6 monoclonal antibody cell 4D-10 strains were prepared by our laboratory.
[0149] 1.2 Experimental animals
[0150] SPF-grade 10-12-week-old BALB / c female mice were purchased from Liaoning Changsheng Biotechnology Co., Ltd.; healthy female rabbits, weighing 2-3 kg.
[0151] 1.3 Immunochromatographic materials
[0152] Nitrocellulose membrane (M135) was purchased from Milipore, USA; glass fiber membrane (GL0194), absorbent paper (H5076), sample pad (GL-b02), PVC board (A-9), connecting tape, and rapid detection plastic card were purchased from Shanghai Jieyi.
[0153] 1.4 Preparation of main reagents
[0154] 1.4...
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