几种动物病毒的基因芯片检测技术

Development of diagnostic genechip for detecting seven kinds of viruses in animals

分别用水疱性口炎病毒、蓝舌病病毒、口蹄疫病毒、猪瘟病毒、牛病毒性腹泻病毒、鹿流行性出血热病毒和赤羽病病毒各一段高度保守的基因片段构建质粒 ,在此基础上制备了芯片探针。提取样品中的核酸 ,经反转录和荧光标记后滴加到芯片上进行特异性杂交 ,对杂交结果扫描检测 ,可同时对上述 7种动物传染病进行快速、准确的诊断 ,此方法敏感性高 ,特异性强 ,适合于大批动物高通量检疫。

The plasmids were developed by means of using respectively high conservative DNA fragments of vesicular stomatitis virus, bluetongue virus, foot-and-mouth disease virus, classical swine fever virus, bovine viral diarrhea virus, epizootic haemorrhagic disease virus of deer and Akabane disease virus. On the basis of the experiment mentioned above, the genechip probs were prepared and spotted on the treated-specially glass slides. The RNA extracted from the samples were reverse-transcripted. The labeled cDNA was subjected to specific hybridizing with the prepared genechip. From the hybridization results detected with photoscanning, the seven viruses mentioned above can be quarantined and identified. This method is not only rapid and sensitive but also exact, which can achieve the goal of large-scale animal quarantine in very short time.