应用ViewRNA技术特异性检测感染细胞中的不同马传染性贫血病毒株

In situ detection of co-infected equine infectious anemia virus heterologous strains in fetal donkey dermal cells using a ViewRNA assay

为了对体外培养细胞感染的不同马传染性贫血病毒(EIAV)株进行特异的鉴别检测,本研究基于一种新的RNA原位杂交-ViewRNA技术,通过设计合成针对EIAVFDDV13和EIAVUK3病毒株基因组RNA的特异性探针,并结合信号放大技术和荧光标记技术,同时通过激光共聚焦显微镜进行观察,建立了可对两种不同株的EIAV基因组RNA进行细胞内原位检测的方法。研究结果表明,两组探针能够特异地与感染细胞中EIAVFDDV13和EIAVUK3的病毒RNA相结合,可在两种不同EIAV株共感染的宿主细胞中实现对两种毒株的有效鉴别和定位检测。该方法为进一步研究EIAV与宿主细胞间相互作用提供了有效手段。

The ViewRNA assay is a novel RNA in situ hybridization method with the high sensitivity to detect even a single RNA copy for specifically differential detection and location of different virus strains in co-infected cell. To detect 2 different strains of equine infectious anemia virus （EIAV） in infected cells, two nucleotide probes were designed and synthesized, of which were capable to hybridize with the genomic RNA of different EIAV strains. The EIAVFDDV13 and EIAVUK3 were co-infected the fetal donkey dermal （FDD） cells and detected by ViewRNA assay under optimized conditions. The results showed that 2 probes were specifically hybridized with the corresponding viral RNA in infected cells and observed under confocal microscrape, indicating that the method was able to effectively identify and locate 2 EIAV different strains in co-infected cells in vitro. The method is competent for further study of the interaction between EIAV strains and host cells.