论文部分内容阅读
目的分析磁微球为载体的H5N1 AIV免疫PCR检测方法。方法模板选择pUC19,需要使用5’端标记的生物素作为引物,实现PCR扩增,进而制备有生物素的DNA分子。固相吸附载体选择磁微球,连接生物素标记的AIV H5N1血凝素蛋白和含有生物素的报告DNA分子,当两者结合之后,会将低含量病毒信号间接放大,建立起更加科学和可靠的检测H5N1 AIV免疫PCR方法。方法确定之后评价和分析PCR方法的灵敏性和特异性、最适报告DNA浓度等。结果采用免疫PCR检测方法可以成功检测到H5NI AIV病毒,确定报告DNA的最适浓度;此种方法也具有比较好的特异性和灵敏性。结论在建立免疫PCR检测方法的过程中以磁微球作为吸附载体,可以在检测H5N1 AIV时具有更高的灵敏性和特异性,是一种比较好的方法。
Objective To analyze the method of H5N1 AIV immunoprecipitation with magnetic microspheres as carrier. Method template selection pUC19, the need to use 5 ’end labeled biotin as a primer, to achieve PCR amplification, and then prepared with biotin DNA molecules. Magnetic beads are immobilized on the solid phase adsorption carrier, and the biotin-labeled AIV H5N1 hemagglutinin protein and the reporter DNA molecule containing biotin are combined. When the two are combined, the virus signal of a low content is amplified indirectly to establish a more scientific and reliable The H5N1 AIV immune PCR assay was used. After the method is established, the sensitivity and specificity of the PCR method are evaluated and analyzed, and the optimal DNA concentration is reported. Results The H5NI AIV virus was successfully detected by the immunological PCR method and the optimal DNA concentration was determined. This method also has good specificity and sensitivity. Conclusion It is a good method to use magnetic microspheres as adsorption carrier during the establishment of immune PCR detection method, which can be used to detect H5N1 AIV with higher sensitivity and specificity.