轮状病毒Vp6蛋白的免疫荧光检测

Immunofluorescence Assay of Rotavirus Vp6 Protein

目的检测在大肠杆菌BL21(DE3)中表达的重组轮状病毒(Rotavirus,RV)Vp6蛋白和RV SA11感染的MA104细胞中不同时间表达的Vp6蛋白及其分布规律。方法应用纯化的重组A组人轮状病毒TB-Chen株Vp6(rVp6)蛋白和RVSA11分别免疫豚鼠,制备抗血清,以抗rVp6豚鼠血清作为检测抗体,应用间接免疫荧光技术检测rVp6蛋白在大肠杆菌BL21(DE3)中的表达。分别用抗Vp6和抗RV SA11豚鼠血清检测Vp6蛋白和RV SA11株在感染的MA104细胞中的分布。结果在轮状病毒感染的MA104细胞和大肠杆菌BL21(DE3)中均能检测到轮状病毒Vp6蛋白。以抗RV SA11血清作为一抗检测感染细胞,比用抗rVp6血清检测具有更强的敏感性。结论免疫荧光试验可用于轮状病毒Vp6蛋白的检测。

Objective To determine the Vp6 protein of rotavims （RV） expressed in E. coli BL21 （DE3）and in RV SAil-infected MAIIM cells as well as the distribution of expressed protein.Metlmtls Immunize guinea pigs with purified recombinant Vp6 （rVp6） protein of TB-Chen strain of human RV group A,exptessed in E. coli BL21（DE3）,and RV SAIl strain respectively. Determine the expression of rVp6 protein in E. coli BL21（DE3） by immunofluorescence assay（IFA）with the sera of guinea pigs immunized with rVp6 protein, and the distribu- tions of Vp6 protein and RV with the sera of guinea pigs immunized with rVp6 protein and RV SAIl strain respectively. Results Vp6 protein was detected in beth E. coli BL21 （DE3） and RV SAil-infected MAIlM cells.The determination of infected cells by IFA with anti-RV SAIl sera was more sensitive than that with anti-rVp6 sera. Conclusion IFA may be used for the determination of Vp6 protein of RV.