利用原位杂交和重组DNA检测禽传染性支气管炎病毒感染
Detection of avian infectious bronchitis viral infection using in situ hybridization and recombinant DNA.
作者信息
Collisson E W, Li J Z, Sneed L W, Peters M L, Wang L
机构信息
Department of Veterinary Microbiology and Parasitology, College of Veterinary Medicine, Texas A&M University, College Station 77843-4467.
出版信息
Vet Microbiol. 1990 Sep;24(3-4):261-71. doi: 10.1016/0378-1135(90)90176-v.
A recombinant DNA probe with specificity for the 3' end of genomic RNA from the Ark 99 strain of infectious bronchitis virus (IBV) was found to hybridize with extracted RNA of three strains with the Ark serotype, as well as the Mass41, Holl52, Gray, JMK, Conn, Fla and SE17 strains of IBV. Viral infection was detected in the cytoplasm of chicken embryo kidney cells inoculated with Mass41, Ark99, SE17 or two recent field isolates of IBV using in situ cytohybridization and a biotinylated probe. In vivo infections were detected in individual cells of tracheas and lungs 2,4, and 6 days after inoculation of chicks with Mass41 and Ark99. In situ hybridization of Ark99 infected tissue sections using 32P-dATP labelled probe indicated that more viral replication was present in the trachea on day 4 than either days 2 or 6; whereas more viral RNA was found in the lungs on day 6 than days 2 or 4 after inoculation.
一种对传染性支气管炎病毒（IBV）Ark 99株基因组RNA 3'端具有特异性的重组DNA探针，被发现可与三种具有Ark血清型的毒株以及IBV的Mass41、Holl52、Gray、JMK、Conn、Fla和SE17株的提取RNA杂交。使用原位细胞杂交和生物素化探针，在接种了Mass41、Ark99、SE17或两种近期IBV野外分离株的鸡胚肾细胞的细胞质中检测到病毒感染。在用Mass41和Ark99接种雏鸡后2、4和6天，在气管和肺的单个细胞中检测到体内感染。使用32P-dATP标记探针对Ark99感染的组织切片进行原位杂交表明，接种后第4天气管中的病毒复制比第2天或第6天更多；而接种后第6天肺中的病毒RNA比第2天或第4天更多。
Zotero 插件