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Newcastle disease is recognized worldwide as the most important infectious diseases of poultry caused by the Newcastle disease virus highly contagious, infectious diseases, can lead to a large number of poultry deaths or the performance of a sharp decline in production, causing huge world poultry industry economic losses. Study the existing problems in the Newcastle disease virus detection method, the use of colloidal gold immunochromatography technology, dual-antibody sandwich principle, the establishment of a rapid detection method of the Newcastle disease virus. This study, rabbits immunized with purified mouse IgG preparation of rabbit anti-mouse serum, rabbit anti-mouse IgG was purified and with a saturated ammonium sulfate salting; two anti-Newcastle Disease Virus (of NDV and then purified by Protein G affinity chromatography) fusion protein (F) monoclonal antibody (anti-NDV-F 6C4 monoclonal antibody, anti-NDV-F monoclonal antibody 4D9), purified by polyacrylamide gel electrophoresis (SDS-PAGE) analysis of purified antibody before and after the effect. The results show that the less the purified antibody heteroaryl protein content and high purity. Trisodium citrate reduction method, successfully prepared 30nm colloidal gold, and its mass to the TEM observation, the result for the diameter of 30nm colloidal gold particles, uniform in size, dispersed state oval. Prepared colloidal gold labeling conditions groping successful establishment of colloidal gold-labeled monoclonal antibody: The optimum pH is 8.7, the antibody, the best combination of an amount of 6.6μg/ml, and prepared a gold labeled NDV- F 6C4 monoclonal antibody. 6C4 of colloidal gold-labeled anti-NDV-F monoclonal antibody preparation immune gold pads, adsorption on glass fiber: 4D9 of anti-NDV-F monoclonal antibody and rabbit anti-mouse IgG, respectively ejection points on the nitrocellulose membrane, the T line. Preparation of double antibody sandwich method, and C lines, prepared as the NDV Immunochromatography rapid detection test strip. After testing, the optimum dilution of the colloidal gold labeled antibody is 1:2 to 30ul/cm sprayed film; T-line 0.6ul/cm monoclonal antibody (3.0mg/ml) the line C 0.6ul/cm rabbit anti hyperimmune serum-mouse IgG (1.5mg/ml) under the conditions of the best detection results. Newcastle disease virus detection test strip, 5min displays the results. Detection of experimentally infected chickens by the specificity, sensitivity, reproducibility and stability test and prove that the result of the method is objective, accurate, and easy to judge the naked eye, and specificity, good reproducibility, simple and fast, is a very the future of the development, promotion and application of new technologies, can be used for epidemiological investigations and field monitoring of the Newcastle disease. The development of the test strip provides a more practical method for the rapid diagnosis of Newcastle disease.
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