蔗糖密度梯度法定量口蹄疫完整病毒粒子(146S)的特异性研究

Research on the Specificity of Sucrose Density Gradient Measurement for the Quantification of FMDV 146S Particles

为验证蔗糖密度梯度法检测口蹄疫完整病毒粒子(146S)的特异性,选择pH值6.0、5.0、4.0 PBS酸解处理3批提纯后的146S灭活抗原液,并将酸解后样品进行146S检测,发现8至11级份OD259峰值消失,146S含量由酸解前的1.77、6.60、3.67μg/mL至酸解后完全不能检测到。同时对3批不含病毒的细胞液按抗原纯化灭活工艺处理后进行146S检测,结果显示3批细胞液在8至11级份均没有明显吸收峰,而且其OD259值与PBS空白对照OD259值基本平行一致,说明不含病毒的细胞液对检测结果没有明显干扰。试验证明,蔗糖密度梯度法定量FMD病毒液中146S含量具有很好的特异性。

In order to verify the specificity of sucrose density gradient measurement for FMDV particles （146S）, three batches of purified 146S inactivated antigen solution were treated with specific acid PBS solution （pH6, 5, 4）, then detected 146S in the acid hydrolysis samples. The results showed that OD259 detect peak of 8 to 11 fractions disappeared, which indicated that 146S didn＇ t exist. But the content of 146S in the samples was 1.77, 6.60 and 3.67μg/mL respectively before acidolysis. At the same time, three batches of cell solution without virus were purified and inactivated. Then 146S was detected with sucrose density gradient technology. The results showed that three batches of cell solution in 8 to 11 fractions had no obvious absorption peak; OD259 values were basically consistent with the blank control, which showed that the cell solution without virus had no effect on the result. It indicated that this method was specific for detecting the amount of 146S in FMDV solution.