Nonidet P-40裂解流感病毒及其对抗体结合影响的透射电镜观察

目的 利用TEM观察经Nonidet P-40处理的A型流感病毒表面形态变化,研究不同表面活性剂-病毒表面相互作用情况,以提供一种较为温和的病毒表面裂解条件,并考察表面活性剂存在条件下对病毒表面抗体结合作用的影响,为利用病毒内部抗体结合病毒下层结构提供免疫诊断学理论基础.方法 通过用不同浓度的非离子型表面活性剂NP-40对完整的A型流感病毒进行处理,经透射电子显微镜成像,观察两种试剂对病毒表面形态的破坏程度,并考察病毒抗体结合情况.结果 NP-40各浓度对病毒表面破坏程度不一,病毒随浓度增高降解程度增大,但在0.1%NP-40及以下浓度上对抗体结合作用影响不大.结论 通过表面活性剂优化处理病毒颗粒,对抗体的结合会产生影响,本研究为利用流感病毒内层抗体结合包膜病毒奠定了样本处理的理论基础.

Transmission electron microscopy investigation of the denudation of the envelope of influenza virus treated with Nonidet P-40 and the binding influence of antibody

Objective Through observing the morphological changes of the prepared influenza viruses (H1N1) treated with the different concentration of Nonidet P-40 solutions and added with antibody against the influenza virus using transmission electron microscopy (TEM), to explore the principle of application of the internal antibody for immunoassay of influenza virus. Methods Through treating the virus samples with serial diluted Nonidet P-40 solutions from 0.01% to 0. 2% and/or not adding antibody against the influenza virus, then investigating the samples by TEM to obtain the morphological changes of the virions. Results The serial images show that the denudation degree of the virions is proportional with the rise of NP-40 concentration, and partly denuded virion image appeared at 0. 1% NP-40 treatment, also under this condition no influence was observed on antibody binding to the virus. Conclusion This work demonstrated the interaction between influenza virion and its antibody under nonionic surfactants existing,which supports the advantages of sample preparation for immunoassay enveloped virus using internal antibody theoretically.