摘要
目的分离并鉴定2010年昆明市无菌性脑膜炎的病原埃可病毒12型（ECH012）病毒株F343／KM／2010，分析其VP1基因遗传特征，为病毒性脑炎的防控提供科学依据。方法采用RD细胞和Hep-2细胞对15份疑似无菌性脑膜炎患者粪便样品进行分离，经微量中和试验对该分离株进行鉴定，通过RT—PCR法扩增病毒VP1基因和测序，并采用MEGA5．05等软件进行进化树和同源性分析。结果9份样品出现细胞病变效应，其中F343／KM／2010分离株经微量中和试验定型为ECH012病毒；经RTPCR法扩增获得873bp的VP1基因，与国内外11株ECH012病毒分离株核苷酸和氨基酸的同源性为81．4％～92．8％和97．6％～99．3％，其中与国内分离株的核苷酸和氨基酸同源性为89．5％～92．8％和98．3%～99．5％。F343／KM／2010株与原型株Travis比较在氨基酸上有8个位点发生突变，而与中国株比较主要在第288位点上发生了突变。在进化树上与国内分离株属于同一个分支，而国外分离株分属其他两个不同的分支。结论F343／KM／2010分离株为肠道病毒ECHO12病毒，与中国其他分离株同属一个分支。
Objective To identify the virus strains isolated from meningitis patients and analyze the gene se quences in Kunming of China in 2010. Methods The feces specimens of suspected meningitis patients were cul- tured in Hep-2 and RD cells to isolate viruses. Viral RNA was extracted and the VP1 gene of Echovirus 12 was amplified using RT PCR and further sequenced. The VP1 gene sequence was compared with other Echovirus 12 from Genbank using MEGAS. 05 software. Results Viruses from 9 clinical specimens were isolated in cell cul ture and F343/KM/2010 isolate was confirmed as Echovirus 12 by microneutralization test. The amplified VP1 gene of Echovirus 12 is 873 nucl.eotides without deletion or insertion. This strain shares 81.4% -92.8% and 97.6%-99.% homology with other Echovirus 12 strains in nucleotide and amino acid sequences, respective- ly; and 89.5%- 92.8%（nueleotides）, 98.3%-99.5% （amino acids） homology compared with strains isola ted from other provinces of China. Amino acid substitutions were found in 8 positions comparing to the Travis strain while amino acid rrmtation was only found at site 288 comparing with the Chinese strain. Phylogenetic analysis grouped our strain with the Chinese strain in the same cluster. Conclusions The Fa43/KM/2010 strain of Echovirus 12 isolated from Yunnan of China was identified as a typical Chinese strain.
出处
《中国病毒病杂志》
CAS
2013年第5期372-376,共5页
Chinese Journal of Viral Diseases
基金
云南省科技计划项目(2013FZ136)
