Hu K F, Morein B, Merza M
Swedish University of Agricultural Sciences, College of Veterinary Medicine, Department of Veterinary Microbiology, Uppsala, Sweden.
J Immunoassay. 1998 May-Aug;19(2-3):209-22. doi: 10.1080/01971529808005481.
Virus neutralization (VN) is an important functional test for evaluating RSV vaccines, also encompassing in mucosal secretion of the respiratory tract considering the infection route. In our previous study, an immunoglobin extraction method described by Bergquist et al. was adopted for RSV ELISA, but it was not suitable for virus neutralization test due to the cell toxicity of the 2% saponin solution used for the antibody extraction. In order to overcome this problem, several solvents including distilled water were tested in the present study for the capacity to extract immunogloblins. Antibodies in the extracts were evaluated and compared by ELISA. Distilled water was as efficient as the 2% saponin solution for extraction of total IgA, RSV specific IgA and IgG. More importantly, the organ extracts obtained subsequently could be used for virus neutralization test without causing adverse effect on the cell culture. Therefore, distilled water was finally chosen as the solvent for immunoglobulin extraction from mucosal organs when both ELISA and virus neutralization test are required.
病毒中和试验(VN)是评估呼吸道合胞病毒(RSV)疫苗的一项重要功能测试,考虑到感染途径,该测试还包括呼吸道黏膜分泌物。在我们之前的研究中,采用了Bergquist等人描述的免疫球蛋白提取方法进行RSV酶联免疫吸附测定(ELISA),但由于用于抗体提取的2%皂苷溶液具有细胞毒性,该方法不适用于病毒中和试验。为了克服这个问题,本研究测试了包括蒸馏水在内的几种溶剂提取免疫球蛋白的能力。通过ELISA对提取物中的抗体进行评估和比较。蒸馏水在提取总IgA、RSV特异性IgA和IgG方面与2%皂苷溶液一样有效。更重要的是,随后获得的器官提取物可用于病毒中和试验,而不会对细胞培养产生不利影响。因此,当需要同时进行ELISA和病毒中和试验时,最终选择蒸馏水作为从黏膜器官提取免疫球蛋白的溶剂。