猪和牛轮状病毒的分离与鉴定

应用MA-104细胞、胰蛋白酶处理培养物和旋转培养技术,成功地从患腹泻仔猪和犊牛的粪便样品中,分离到6株猪轮状病毒和4株牛轮状病毒。经形态学、血清学和病毒RNA电泳分析等鉴定,确为典型轮状病毒。经MA-104细胞连续传代,这10株轮状病毒均成为细胞适应毒株,并在该细胞上产生明显的细胞病变。

ISOLATION AND IDENTIFICATION OF PORCINE AND BOVINE ROTAVIRUS

Six porc ine rotavirus strains (designated PRV1 ,PRV8,PRVll, PRV2,PRVl5 PRV17) and four bovine rotavirus strains ( BRV6551 ,BRV6555, BRV6571, BRV 6576 ) were successfully isolated and adapted in MA-104 cells from the feces of piglets and calves with diarrhea. MA-104 cells with trypsin and in rolling culture tubes was the system of choice for virus replication and isolation. All of the isolated porcine and bovine rotavirus strains produced obvious cylopathic effects in MA-104 cells.EM revealed the presence of particles with the typical rotavirus morphology in PRV8 and BRV6555 isolates.All of the isolated porcine and bovine rotavirus strains were antigenically related to each other and to the reference bovine rotavirus strain NCDV by IF, ELISA, and ID. RNA electrophoresis analysis indicated that the isolated porcine and bovine rotavirus were all resolved out 11 clear bands in. polyacry?lamide gel.But the electrophoretic profiles of the isolated porcine and bovine rotavirus strains had their own characteristics which were different from the reference rotavirus strains NCDV,pocine S80,and human Wa.And two different electrophoresis profiles appeared both among the six porcine (PBV1 and other five) and the four bovine rotavirus strains (BRV6576 and other three). 12(50%) of 24 fecal samples from piglets with diarrhea and 16(55%) of 29 fecal samples from calves with diarrhea were positive by ELISA direct (sandwich) assay.