A colloidal gold immunochromatographic test strip for rapid detection of Edwardsiella tarda
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A slow Edward's, immunochromatographic detection technology, used in measurement devices, analytical materials, instruments, etc.
Inactive Publication Date: 2011-12-21
福建省淡水水产研究所
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Due to its fast, convenient, no need for special equipment, intuitive results and other advantages, this method has developed rapidly in recent years and has been widely used in the field of biomedicine, especially in medical testing. However, products used in the field of animal disease prevention and control detection less
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Embodiment 1
[0017] Embodiment 1, establishment of anti-Edwardia tarda monoclonal antibody cell line
[0018] Prepare the inactivated antigen of Edwardsiella tarda, immunize Balb / c mice, prepare spleen cell suspension after immunization; prepare myeloma cell suspension at the same time; perform cell hybridization and fusion of spleen cells and myeloma cells, and screen positive monoclonal cell wells ; After further positive cloning and screening, two anti-Edwardia tarda monoclonal antibody hybridoma cell lines were established, named ETMcAb3A7 and ETMcAb4F11 respectively. The two cell lines were deposited in China Center for Type Culture Collection (Wuhan, China, Wuhan University) on June 6, 2010, and the preservation numbers are CCTCC NO: C201061 and CCTCC NO: C201062.
Embodiment 2
[0019] Example 2, Preparation of anti-Edwardella tarda monoclonal antibody ascites
[0020] Anti-Edwardia tarda monoclonal antibody cell lines ETMcAb3A7 and ETMcAb4F11 were cultured, monoclonal antibody cells were collected, Balb / c mice were injected intraperitoneally, and raised in a clean environment. After about 7-10 days, observe the abdomen of the mice, and those with obvious ascites use 12 # The needle is punctured from the lower side of the abdominal cavity to recover ascites, 3000r·min -1 Centrifuge for 5 minutes to precipitate red blood cells and blood cells, collect the light yellow ascites in the upper layer, and save it for later use.
[0022] Prepare the inactivated antigen of Edwardsiella tarda, add the same amount of complete Freund's adjuvant for the first immunization, inject New Zealand rabbits, perform the second immunization within 2-3 weeks, add the same amount of incomplete Freund's adjuvant to the antigen, and then immunize for the third time after one month . After the immunization, blood was collected through the carotid artery, and after 5000r·min -1 Serum was collected after centrifugation, aliquoted aseptically, and stored at -80°C for future use.
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Abstract
A colloidal gold immunochromatographic test strip for rapid detection of Edwardsiella tarda, the invention relates to a method of using nano-colloidal gold particle-labeled antibody technology and double-antibody sandwich immunochromatography to detect antigen patterns, and prepared a colloid of Edwardsiella tarda Gold immunochromatographic test strips. A colloidal gold immunochromatographic detection test strip for Edwardsiella tarda and its preparation method and application are provided. The test strip contains sample pads, gold standard pads, nitrocellulose membranes with detection line and quality control line antibodies, absorbent pads, and bottom plates. The sample pads, gold standard pads, nitrocellulose membranes, and absorbent pads are sequentially overlapped and connected. Assembled by pasting on the bottom plate. The detection application of the test strip is simple in operation, good in specificity, high in sensitivity, convenient and fast, does not require special equipment and professional training, and the results are clear and easy to distinguish and easy to promote. It is suitable for detection of infection and contamination of Edwardsiella tarda.
Description
technical field [0001] The invention belongs to the field of biological detection, in particular to a colloidal gold immunochromatographic test strip for rapid detection of Edwardsiella tarda and its preparation and application in the detection of Edwardsiella tarda. Background technique [0002] Edwardsiella tarda is an explosive type of bacterial disease in aquaculture fisheries. It is prevalent in commercially farmed fish such as Forktail family, Eel family, flounder and carp. It mainly causes liver and kidney diseases of fish. The disease occurs mainly in high water temperature periods such as spring and summer, autumn, etc., and reaches a peak in summer. Its prevalence is wide, the mortality rate is high, and the harm is very serious. It has attracted great attention from the aquaculture industry. The detection and identification of Edwardsiella tarda is currently mainly limited to bacteriological examination or pathological observation. The conventional isolation, iden...
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