论文部分内容阅读
目的检测乙脑病毒SA14-14-2减毒株的病毒外源因子。方法用乙脑病毒免疫血清(阳性)将乙脑病毒SA14-14-2减毒株中和后,采用中和后的样品分别感染指示细胞(3T3、NRK、C3/36细胞),直接培养观察及红细胞吸附试验;中和后样品感染小鼠细胞系(3T3)培养14d的上清液再接种指示细胞(MRC-5、Vero、BHK21、3T3)培养观察及红细胞吸附试验;同时将流感病毒接种MDCK细胞,作为试验阳性对照。结果乙脑病毒SA14-14-2减毒株中和后的样品接种指示细胞(3T3、NRK、C6/36细胞),直接培养观察7 d、14 d,细胞形态正常,无细胞病变征兆,红细胞吸附试验为阴性;中和后样品感染小鼠细胞系(3T3)培养14 d的上清液再接种指示细胞(MRC-5、Vero、BHK-21、3T3)培养观察7 d、14 d,细胞形态正常,无细胞病变征兆,红细胞吸附试验为阴性。结论乙脑病毒SA14-14-2减毒株中未检测到人源、猴源、鼠源及其他病毒的污染,符合《WHO Technical Report Series,No.910,2002》的质量标准,可安全地用于乙脑减毒活疫苗的生产。
Objective To detect the virus exogenous factor of JE virus attenuated strain SA14-14-2. Methods Neutralization of JEV SA14-14-2 attenuated strain with JEV immune serum (positive) was used to infect the indicator cells (3T3, NRK, C3 / 36 cells) And erythrocyte adsorption assay. After neutralization, the supernatant of the mouse cell line (3T3) infected with 3T3 cells was cultured and observed for erythrocyte adsorption (MRC-5, Vero, BHK21 and 3T3) MDCK cells as a positive control test. Results After neutralization with JEV attenuated strain SA14-14-2, the indicator cells (3T3, NRK and C6 / 36 cells) were inoculated directly and observed for 7 d and 14 d. The cells were normal in morphology and showed no sign of cytopathic effect After neutralization, the supernatant of mouse cell line (3T3) infected with 3T3 cells was cultured for 7 days and 14 days after being inoculated with indicator cells (MRC-5, Vero, BHK-21 and 3T3) Normal shape, no signs of cytopathic effect, red blood cell adsorption test was negative. Conclusion No contamination of human, monkey, mouse and other viruses was detected in attenuated strains of JE virus SA14-14-2, which met the quality standards of “WHO Technical Report Series, No.910, 2002” For JE attenuated live vaccine production.