核酸探针原位杂交检测白斑综合症病毒的组织特异性  

A STUDY ON TISSUE TROPISM OF WHITE SPOT SYNDROME VIRUS(WSSV)BY IN SITU HYBRIDIZATION

作　　者：吕玲[1] 何建国[1] 邓敏[1] 翁少萍[1] 左涛 

机构地区：[1]中山大学生命科学学院,广东广州510275

出　　处：《热带海洋》2000年第4期86-91,共6页

基　　金：国家自然科学基金资助项目 (3 9670 5 77) ;广东省自然科学基金资助项目 (960 0 70 )

摘　　要：从白斑综合症病毒 (whitespotsyndromevirus ,简称WSSV)部分基因组文库中得到核酸探针 ,采用原位杂交检测方法检测对虾甲壳下表皮、胃、后肠、鳃、触角腺的上皮细胞、淋巴器官的基质细胞、肌细胞、心肌细胞及结缔组织细胞 ,原位杂交结果均为阳性。对虾甲壳下表皮的上皮细胞、胃的上皮细胞对病毒较其它部位敏感 ,感染程度高 ;中肠上皮细胞、肝胰腺上皮细胞、淋巴器官内皮细胞未发现感染病毒 ,原位杂交用于WSSV检测 ,比组织切片HE染色更准确、更敏感。在同一组织的同一部位 ,原位杂交所得到的阳性细胞个数比HE多。探针大小不同 ,敏感性也有变化。短的探针 41 3bp敏感性较高 ,长的探针敏感性降低。A WSSV(white spot syndrome virus)DNA fragment from the WSSV genomic library constituted by us was used as a probe to detect WSSV by in situ hybridization.It was found that WSSV in some cells of the infected Penaeus monodon ,such as epithelial cells of integument,stomach,gills,hindgut,antennal gland,stromal matrix cells of lymphoid organ,muscle cells,myocardial cells and connective tissue cells,were positive.The epithelial cells of stomach and integument were more sensitive to WSSV,while negative reactions were observed in the epithelial cells of hapatopancreas tissue,midgut,and the endothelial cells of lymphoid organ.The results showed that the method of in situ hybridization to detect WSSV was more accurate and sensitive than the classical method by HE stain,and more positive cells were obtained.The sensibility of in situ hybridization varied with the size of the DNA probes. The probe of 413bp fragment from WSSV was more sensitive than 941bp and 1?447bp.

关 键 词：白斑综合症病毒 核酸探针 原位杂位 检测 对虾 

分 类 号：S945.462[农业科学—水产养殖]