Kit for detecting prion and preparation method thereof
A technology of prions and kits, applied in the biological field, can solve a handful of problems and achieve rapid and accurate detection, high sensitivity, and convenient use
Active Publication Date: 2014-04-09
龙佑(泰安)医学科技有限公司
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Problems solved by technology
Although many countries in the world have carried out different degrees of research on the diagnostic methods of BSE, such as enzyme-linked immunosorbent assay (ELISA), immunohistochemical detection, histopathological detection, experimental animal inoculation detection, histochemical detection, etc., but the detection of BSE There are only a handful of commercial kits in the world. At present, there is no rapid and sensitive scientific detection method for BSE in China.
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[0066] A preparation method of a kit for detecting prions, comprising:
[0067] (1) The preparation method of the ELISA plate coated with the capture antibody, the steps are as follows:
[0068] A. Antigen preparation: Purify prion protein molecules (PrPSc) from the brain tissue of prion-infected hamsters;
[0069] B. Immunization: mix the purified PrPSc with Freund's adjuvant, and subcutaneously inject Balb / c mice;
[0070] C. Preparation of cloned hybridoma cells: After two months, the spleen of the mouse was taken, and a single splenocyte was collected for fusion with SP2 / 0 myeloma cells to prepare hybridoma cells, and the hybridoma cells that could secrete anti-PrPSc antibodies were screened, expanded and cultured, that is monoclonal hybridoma cells. Different monoclonal hybridoma cells secrete antibodies against different epitopes.
[0071] D. Preparation of monoclonal antibody: Inject the monoclonal hybridoma cells into the peritoneal cavity of the mouse, causing the ...
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The invention relates to a kit for detecting prion and a preparation method thereof. The kit comprises an ELISA (enzyme-linked immunosorbent assay) plate coated with a capture antibody, a detection antibody combined with the prion and coupled with a biotin labeling, as well as antibody diluent, sample diluent, washing liquid, streptavidin-HRP, substrate solution and stop solution, wherein the capture antibody is a capture antibody combined and reacted with an amino acid sequence shown in SEQ ID NO.1; and the detection antibody combined with the prion and coupled with the biotin labeling is a detection antibody combined and reacted with an amino acid sequence shown in SEQ ID NO.2 shown. According to the invention, the kit for detecting the prion is established through the utilization of the ELISA technology, and has the advantages of convenience for use, high sensitivity, capability of quickly and accurately detecting the prion, simplicity in preparation, low cost and facilitation of popularization and industrial production.
Description
technical field [0001] The invention relates to a kit for detecting prions and a preparation method thereof, belonging to the technical field of biotechnology. Background technique [0002] Prion, also known as protein infection factor, is a small molecule non-immune hydrophobic protein that can infect animals and replicate in host cells. Prion is an old name for protein, and prion means protein virus. [0003] It is a kind of infectious pathogenic factor that can cause central nervous system lesions in mammals and humans. Among them, the pathogen of mad cow disease that "talks about the discoloration of cows" belongs to prions. Mad Cow Disease, or Bovine Spongiform Encephalopathy (BSE), is a type of transmissible spongiform encephalopathy. The symptoms are: the central nervous system of most sick cattle changes, abnormal behavior, restlessness, and unsteady gait. After dissection, it was found that the gray matter of the central nervous system of the sick cattle formed ...
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IPC IPC(8): G01N33/577
Inventor 王传华刘芸
Owner 龙佑(泰安)医学科技有限公司