通用型病毒检测芯片在三种人类致病病毒属检测中的应用

Universal viral detection microarray for detecting three human pathogenic viruses

目的研究制备对人类有致病作用的38个属的病毒寡核苷酸(oligo)基因芯片对三种人类致病病毒的检测能力。方法应用生物信息学软件设计70-meroligo探针,固定于玻片载体制备成基因芯片。以痘苗病毒天坛株、甲肝病毒、乙肝病毒作为检测样本,提取病毒核酸,经随机引物扩增、荧光标记,用于芯片杂交,清洗和干燥后对芯片进行扫描和数据分析。结果芯片上oligo探针能与相应病毒的PCR扩增样品杂交,呈现阳性荧光信号。结论建立的病毒寡核苷酸基因芯片能够检出和区分三种检测病毒,为进行未知病毒的基因芯片筛查方法的建立奠定了基础。

Objective To evaluate the established high throughput oligo microarray with three human pathogenic viruses, including Orthopoxvirus, Orthohepadnavirus and Hepatovirus. Methods Using a bioinformatics software, pathogenic viruses genomes were analyzed and 70-mer oligo-nucleotide probes were designed and synthesized. The oligo microarray was prepared by micro-girding. Nucleotide segments derived from virus genomes were labeled by the RoundA/B/C random amplification technology. The labeled nucleotide samples were subsequently applied to the oligo microarray for hybridization, which followed the analysis by signal scanning. Results Viral nucleotides were able to hybridize to most expected probe spots on the microarray, revealing positive fluorescent signals. Conclusion The established oligo microarray for pathogenic viruses can identify and distinguish the three tested viruses, supplying a reliable scientific basis for screening unknown pathogenic virus with the technology of oligo microarray.