岱山县新型布尼亚病毒分离与鉴定  

Isolation and molecular identification of novel Bunyavirus in Daishan county

作　　者：夏仕军[1] 王忠发[1] 李世波[2] 任宜[1] 虞吉寅[1] 张博[1] 

机构地区：[1]岱山县疾病预防控制中心,浙江岱山316200 [2]温州医科大学附属舟山医院,浙江温州316000

出　　处：《中国卫生检验杂志》2014年第3期368-371,共4页Chinese Journal of Health Laboratory Technology

基　　金：2012年浙江省医药卫生平台重点资助计划(2012-ZDA044);2013年舟山市卫生局攻关项目(2013G01)

摘　　要：目的应用非洲绿猴肾(Vero)细胞分离培养发热伴血小板减少综合征病人血清中的新型布尼亚病毒,采用病毒全基因组测序及序列比对技术进行分析鉴定。方法选取2012年岱山县经荧光RT-PCR检测阳性的发热伴血小板减少综合征病人急性期血清6份在Vero细胞系中培养。用RT-PCR测定新型布尼亚病毒L、M、S三个特异性基因片段并对上述扩增产物进行测序,测序结果与GenBank公布的序列进行比对确认。用DNAStar软件构建进化树与序列距离表。结果从6份血清样本中分离到2株新型布尼亚病毒,通过基因组序列比对鉴定为新型布尼亚病毒,病毒株送浙江省疾控中心病毒所复核得到确认。结论 Vero细胞可用于新型布尼亚病毒分离培养,采用分子鉴定技术鉴定新型布尼亚病毒具有简单、快速、准确的良好效果,适用于基层疾控中心病毒分离与鉴定工作的开展。Objective To culture and isolate novel Bunyavirus from serum of severe fever patients accompanying by thrombocy- topenia syndrome （SFTS） using Veto cell line, so as to analyze and identify the virus by viral whole - genome sequencing and sequence alignment techniques. Methods SFTS patients were detected by fluorescent RT - PCR, and 6 serum specimens were selected from acute phase of these cases in Daishan in 2012 for culture in Veto cell line. The specific fragments L, M and S of novel bunyavirus were determined by RT - PCR, and the amplification products were sequenced, then the results were compared with sequence in GenBank. The phylogenetic trees and sequence distance table were generated by DNAStar software. Results Two strains of novel Bunyavirus were isolated from the 6 SFFS serum specimens and identified as novel Bunyavirus by genome sequence alignment. The virus strains then were confirmed by Zhejiang Provincial Center for Disease Control and Pre- vention. Conclusion Veto cell line is available for culture and isolation of novel Bunyavirus. It is easy, fast and accurate for identifying novel Bunyavirus by molecular biology technique, which is suitable for isolation and identification of virus in grass - roots CDC.

关 键 词：新型布尼亚病毒 VERO细胞 病毒培养 分子鉴定 

分 类 号：R373.9[医药卫生—病原生物学]