关键词: 鹿流行性出血热病毒, 阿卡斑病毒, 蓝舌病病毒, 水泡性口炎病毒, 液相芯片检测技术

Abstract: The aim is to develop liquichip technique to detect infection of epizootic haemorrhagic disease virus of deer (EHDV), akabane virus (AKV), bluetongue virus (BTV), and vesicular stomatitis virus (VSV). The VP7 gene of EHDV, N gene of AKV, VP7 gene of BTV and NP gene of VSV in the GenBank were analyzed by using the software DNAStar 5.0. Specific gene probes of all these four viruses labeled with biotin were prepared and coupled with fluorescencecoded microspheres. The probes were used for hybridization reaction to RTPCR products of the four viruses, and then the liquichip detection technique for detection of all these four viruses was established by using liquichip 200 to detect fluorescence signals in the reaction system. The results showed that this assay method displayed better specificity to RTPCR products of correspondent viruses and no response to that of other insectborne disease viruses when being used for detection. The sensitivity test indicated that the detecting limitation for the viruses could reach to 50100 TCID50. The rapidly high throughput liquichip detection technique to detect the four insectborne disease viruses was established, which provided a foundation and exploration for further research to detect other viruses with the same technique in animal husbandry.

Key words: epizootic haemorrhagic disease virus of deer, akabane virus, bluetongue virus, vesicular stomatitis virus, liquichip detection technique