摘要
通过12%SDS-PAGE和5%～20%梯度SDS-PAGE二次制备电泳纯化浙贝母花叶病毒(Thunberg fritillary mosaic virus,TFMV)的外壳蛋白,4次免疫小鼠,获高纯度的抗血清。分别用Western blot、间接ELI SA、免疫扩散、火箭免疫电泳和对流免疫电泳5种免疫检测技术对TFMV样品进行了检测。结果表明,间接ELISA方法不能灵敏地检测新鲜病汁液中的病毒粒子,免疫扩散不能用于未变性病毒粒子的检测。Western blot灵敏度高,但操作复杂。火箭免疫电泳和对流免疫电泳均具有操作简单、灵敏度高、检测周期短的优点,但前者需抗血清的量大。5种免疫检测技术中,对流免疫电泳最适合浙贝母病样中TFMV的快速检测。
The coat protein of Thunberg fritillary mosaic virus （TFMV） was purified by 12%SDS-PAGE and 5%-20% gradient SDS-PAGE. High quality antiserum was obtained by immunization of mice with the purified virus preparation for four times. Western blot, immunodiffusion, indirect ELISA, counterimmunoelectrophoresis and rocket immunoelectrophoresis techniques were used for the detection of TFMV in different samples. It was demonstrated that indirect ELISA was not suitable for the detection of TFMV in virus detection in fresh infected plant sap. The nature viral particles can not be detected by immuodiffusion. Western blot was sensitive but it was complicate in operation. Counterimmunoelectrophoresis and rocket immunoelectrophoresis were simple, sensitive and timesaving. However, rocket immunoelectrophoresis was greatly antiserum consuming, Counterimmunoelectrophoresis is the best serological method for the detection of TFMV in all five methods.
出处
《科技通报》
2006年第4期506-509,518,共5页
Bulletin of Science and Technology
基金
浙江省自然科学基金(ZA0207)
国家自然科学基金项目(30200008)
国家杰出青年基金项目(30225031)
