To establish an indirect competitive enzyme-linked immunosorbent assay(ic-ELISA)method for the detection of zilpterol(ZIL)residues,in this study,based on the obtained rabbit anti-ZIL polyclonal an-tibody serum and the prepared complete antigen as the coating source,an ic-ELISA method for the detec-tion of ZIL residues was established by optimizing the reaction conditions;The results showed that the best dilution ratio of rabbit serum was 1:6 000,and the best antigen coating concentration was 2 μg/mL,the half inhibition concentration(IC50)is 0.317 ng/mL,the linear detection range(IC20-IC80)is 0.136-1.181 ng/mL,and the minimum detection limit(LOD,IC10)is 0.081 ng/mL.After optimization,the optimal pH of the detection system is 7.4,the salt concentration is≤0.4 mol/L,the content of methanol,ethanol,ace-tone and DMSO is≤5％,the content of acetonitrile is≤10％,and the concentration of bovine urine in the detection matrix is≤15％.The detection method has good stability and specificity.The addition recovery rate of intra-batch and inter-batch repeated test samples is 101％-1 12％and 101％-120％respectively,and the coefficient of variation is less than 10％,The correlation coefficient between the test results and HPLC-MS test results is R2=0.990 6.The detection method is fast,sensitive,accurate,reproducible and low detec-tion limit,and is suitable for high-throughput detection and on-site rapid screening of ZIL residues.

为制备齐帕特罗(ZIL)完全抗原,获得抗ZIL多克隆抗体(pAb),将ZIL与4-溴丁酸乙酯反应后,采用EDC/NHS方法使ZIL-丁酸盐中间体与牛血清白蛋白(BSA)和鸡卵清蛋白(OVA)进行偶联,分别用紫外全波长扫描(UV)、聚丙烯酰胺凝胶电泳(SDS-PAGE)进行鉴定后,通过免疫新西兰大白兔获得抗ZIL多抗血清,并对抗体效价、半数抑制浓度(IC50)和特异性进行鉴定.HPLC-MS检测结果显示,改造后的主产物分子质量为348.21 u,约占整个反应体系产物的92.23％,与ZIL-丁酸盐分子质量完全相符;SDS-PAGE电泳结果显示,BSA/OVA在电泳中的迁移速率明显大于偶联产物;UV结果显示,相比载体蛋白,偶联产物ZIL-BSA/OVA的特征吸收峰均发生了明显变化;ELISA效价检测结果表明,ZIL多抗血清效价在1:6400以上,通过建立标准曲线测得IC50分别为0.38 ng/mL和1.13 ng/mL,并且与沙丁胺醇(SAL)、特布他林(TBL)、溴布特罗(BRO)、西马特罗(CIM)、克仑特罗(CLB)、盐酸多巴胺(DH)、莱克多巴胺(RAC)、达氟沙星(DAN)、泰乐菌素(TYL)均不存在任何交叉反应.成功制备了ZIL完全抗原ZIL-BSA/OVA,获得了一种高灵敏度和高特异性的抗ZIL pAb,为后续建立ZIL免疫学快速检测方法奠定了基础.

免疫系统中基因的精确表达和调控是生物体产生对病原体强大免疫能力及限制自身免疫的重要因素.越来越多的研究表明,长链非编码RNA(lncRNA)与免疫功能密切相关,能够通过调控免疫细胞分化发育以及免疫细胞效应功能参与免疫反应的调节.论文结合lncRNA调控作用的分子机制,阐述相关lncRNA在免疫应答和免疫调节过程中发挥的生物学效应,以期为从事lncRNA调控免疫应答机制研究提供参考.