利用基因芯片技术对一种未知病毒的筛查与鉴定  

The screening and identification of an unknown virus by DNA microarray

作　　者：杨银辉[1] 朱晓光[1] 张永国[1] 刘伯华[1] 康晓平[1] 刘洪[1] 祝庆余[1] 

机构地区：[1]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071

出　　处：《中华检验医学杂志》2007年第12期1360-1363,共4页Chinese Journal of Laboratory Medicine

摘　　要：目的利用本研究室已建立的人类医学病毒属水平基因芯片筛查技术及主要虫媒病毒基因芯片检测技术,对2006年7月从山西临沂地区猪脑组织中分离到的未知病毒进行筛查与鉴定,确定其病原体。方法从猪脑组织中分离到病原体,接种金黄地鼠肾细胞(BHK21细胞),能产生细胞病变(cytopathic effect,CPE)。收集病毒感染的细胞上清,提取 RNA,经逆转录及随机引物 PCR扩增后,与医学病毒属水平基因芯片进行杂交,初步确定其所在的病毒属之后,再与主要虫媒病毒基因检测芯片进行杂交,进一步将病毒鉴定到种。通过与其他鉴定结果相比较,确定其检测特异性。结果与医学病毒属水平基因芯片杂交结果显示,该病原体属于黄病毒属病毒。结合流行病学调查分析,该病毒可能是一种虫媒病毒,因此进一步与主要虫媒病毒基因芯片杂交,基本可以判断该病原体为乙型脑炎病毒,与本室 PCR 及基因序列测定结果一致。结论本研究室所建立的基因芯片筛查与鉴定技术可初步应用于对未知病毒的鉴定,为未知病毒性病原体的确定提供了一种新的技术方法。Objective In our previous study, we established DNA microarray technology for identification of medical viruses on genus levels and arboviruses on species levels. In this study, we employed these microarrays to determine the pathogen of newly isolated unknown virus in July, 2006 from pig brain in Shanxi province. Methods The pathogen isolated from pig brains were inoculated in BHK21 cells. After CPE were observed, the supernatants were collected and RNA was extracted. After reverse transcription and random PCR amplification, the labeled nucleic acids were hybridized with DNA microarrays. Results The hybridization results with medical viruses DNA microarray indicated that the unknown virus belonged to Flavivirus. Combined with epidemiological investigation, we presumed that it might be a kind of arbovirus. Then the labeled specimen were further hybridized with arbovirus DNA microarray and the results confirmed that it was Japanese encephalitis virus （JBV）. This coincided with PCR and sequencing analysis. Conclusions The DNA microarray we established previously could be employed to identify unknown viruses. This method provides a new method for determining new viral pathogens.

关 键 词：微阵列分析 脑炎病毒 流行病学 分子 

分 类 号：R450[医药卫生—治疗学]