一起病毒性脑炎病因的分子生物学鉴定

Molecular-biological identification of pathogens which caused an outbreak of viral encephalitis in Jinan area

目的探讨2003年山东济南地区病毒性脑炎(VE)流行病因。方法通过随机PCR及肠道病毒特异性的RT-PCR扩增获得病毒分离株特异性核酸片段,测序并进行同源性分析和多序列比对,确定病毒种类及分型。结果从患者标本中分离到5株病毒,以随机PCR获得其中1株病毒的核酸片段,经BLAST分析发现其与肠道病毒同源性最高,然后分别测得5株病毒5′端非编码区及部分VP1区核酸序列,经序列比对分析,确定5株病毒均为小RNA病毒科肠道病毒属的柯萨奇B5,并且均与2002—2004年间浙江无菌性脑膜炎流行期间分离的柯萨奇B5分离株同源性最高(95%以上)。结论柯萨奇B5是2003年济南地区VE流行的重要病因之一。

Objective To investigate the etiology of the outbreak of viral encephalitis in Jinan area in 2003.Methods Virus-specific nucleic acid fragments were amplified by random PCR and RT-PCR using specific primers to enterovirus. After sequencing, the gene sequence was handled by the program BLAST for homologous analysis and the software Clustal W 1.82 for multiple sequence alignment to identify the etiology and its genotype, Results Five strains were isolated from clinical specimens. A gene fragment for one strain was acquired using random PCR, which was highly homologous to enterovirus. Then, the 5＇ non-translated region and partial VP1 region were amplified and sequenced. The five isolated strains were all identified as Coxsackievirus B5, and what was more, they were most homologous to the strain isolated during the outbreak of aseptic meningitis and encephalitis in Zhejiang province from 2002 to 2004. Conclusion Coxsackievirus B5 is closely associated with the outbreak of viral encephalitis in Jinan area in 2003. It is an important etiology but other viruses may also played a role which remains to be clarified.