中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2010年
5期
438-443
,共6页
姜红月%李铭华%杨小兵%李培龙%张静%孟凤霞%郭玉红%梅立志%付士红%贺圆圆%王环宇%王子军%杨维中%王志玉%梁国栋
薑紅月%李銘華%楊小兵%李培龍%張靜%孟鳳霞%郭玉紅%梅立誌%付士紅%賀圓圓%王環宇%王子軍%楊維中%王誌玉%樑國棟
강홍월%리명화%양소병%리배룡%장정%맹봉하%곽옥홍%매립지%부사홍%하원원%왕배우%왕자군%양유중%왕지옥%량국동
脑炎病毒亚组,日本%基因型%序列分析%种系发生
腦炎病毒亞組,日本%基因型%序列分析%種繫髮生
뇌염병독아조,일본%기인형%서렬분석%충계발생
Encephalitis viruses,Japanese%Genotype%Sequence analysis%Phylogeny
目的 对2008年湖北省新分离的2株乙型脑炎(简称乙脑)病毒进行全基因组序列测定和分析,了解当地乙脑病毒株的分子生物学特性.方法 病毒复苏鉴定后,使用覆盖乙脑病毒全基因组的16对特异性引物,RT-PCR法扩增乙脑病毒HBZG08-09株和HBZG08-55株的全基因组片段,直接测序拼接获得全基因组序列.使用ClustalX 1.83、MegAlign、Mega 4和Genedoc 3.2等生物学软件进行序列比对、核苷酸和氨基酸同源性分析、系统进化分析及氨基酸位点差异分析.结果 新分离的2株乙脑病毒基因组全长均为10 965个核苷酸,从97位到10 392位为开放阅读框,编码3432个氨基酸,2株间的全基因组核苷酸和氨摹酸间源性分别是98.2%和99.7%.进一步的基因型研究显示HBZG08-09株和HBZG08-55株均属于基因Ⅰ型乙脑病毒.2株病毒与我国近年来在河南及浙江的乙脑病毒分离株进化关系最近.与目前使用的减毒活疫苗株SA-14-14-2相比较,HBZG08-09株全基因组共存在82个氨基酸差异,HBZG08-55株存在84个氨基酸差异.但影响毒力或抗原的关键氨基酸位点未发生改变.结论 新分离的2株乙脑病毒均属于基因Ⅰ型,与疫苗株相比关键氨基酸位点未见变异.
目的 對2008年湖北省新分離的2株乙型腦炎(簡稱乙腦)病毒進行全基因組序列測定和分析,瞭解噹地乙腦病毒株的分子生物學特性.方法 病毒複囌鑒定後,使用覆蓋乙腦病毒全基因組的16對特異性引物,RT-PCR法擴增乙腦病毒HBZG08-09株和HBZG08-55株的全基因組片段,直接測序拼接穫得全基因組序列.使用ClustalX 1.83、MegAlign、Mega 4和Genedoc 3.2等生物學軟件進行序列比對、覈苷痠和氨基痠同源性分析、繫統進化分析及氨基痠位點差異分析.結果 新分離的2株乙腦病毒基因組全長均為10 965箇覈苷痠,從97位到10 392位為開放閱讀框,編碼3432箇氨基痠,2株間的全基因組覈苷痠和氨摹痠間源性分彆是98.2%和99.7%.進一步的基因型研究顯示HBZG08-09株和HBZG08-55株均屬于基因Ⅰ型乙腦病毒.2株病毒與我國近年來在河南及浙江的乙腦病毒分離株進化關繫最近.與目前使用的減毒活疫苗株SA-14-14-2相比較,HBZG08-09株全基因組共存在82箇氨基痠差異,HBZG08-55株存在84箇氨基痠差異.但影響毒力或抗原的關鍵氨基痠位點未髮生改變.結論 新分離的2株乙腦病毒均屬于基因Ⅰ型,與疫苗株相比關鍵氨基痠位點未見變異.
목적 대2008년호북성신분리적2주을형뇌염(간칭을뇌)병독진행전기인조서렬측정화분석,료해당지을뇌병독주적분자생물학특성.방법 병독복소감정후,사용복개을뇌병독전기인조적16대특이성인물,RT-PCR법확증을뇌병독HBZG08-09주화HBZG08-55주적전기인조편단,직접측서병접획득전기인조서렬.사용ClustalX 1.83、MegAlign、Mega 4화Genedoc 3.2등생물학연건진행서렬비대、핵감산화안기산동원성분석、계통진화분석급안기산위점차이분석.결과 신분리적2주을뇌병독기인조전장균위10 965개핵감산,종97위도10 392위위개방열독광,편마3432개안기산,2주간적전기인조핵감산화안모산간원성분별시98.2%화99.7%.진일보적기인형연구현시HBZG08-09주화HBZG08-55주균속우기인Ⅰ형을뇌병독.2주병독여아국근년래재하남급절강적을뇌병독분리주진화관계최근.여목전사용적감독활역묘주SA-14-14-2상비교,HBZG08-09주전기인조공존재82개안기산차이,HBZG08-55주존재84개안기산차이.단영향독력혹항원적관건안기산위점미발생개변.결론 신분리적2주을뇌병독균속우기인Ⅰ형,여역묘주상비관건안기산위점미견변이.
Objective To sequence and analyze the complete genome of two new Japanese encephalitis virus (JEV) strains isolated from mosquitoes collected in Hubei province in 2008, and to understand the molecular biological characteristics of JEV in this area. MethodsRT-PCR was used to amplify the fragments of HBZG08-09 strain and HBZG08-55 strain with 16 pairs overlapping primers after they had been recovered and identified, then the full-length genome was obtained by sequencing and splicing. Biological sequence alignment, nucleotide and amino acid sequence analysis, phylogenetic analysis and analysis of amino acid differences were pedormed by the software of ClustalX (1.83), MegAlign, Mega (4.0) and Genedoc (3.2). ResultsThe genome of two new strains were both 10 965 nucleotides in length with a single open reading frame from 96 to 10 392 coding for a 3432 amino acid poly-protein,the homology of nucleotide and amino acid sequence between two isolates were 98. 2% and 99.7% respectively. Further study showed that the new strains were both belonging to genotype Ⅰ. Two new strains were most closely related to isolates obtained from Henan and Zhejiang province in recent years. Compared with the live attenuated vaccine strain SA-14-14-2 in China, HBZG08-09 strain had 82 amino acid divergence; HBZG08-55 had 84 amino acid divergences. But the amino acid difference oceurred in sites were not the key ones affecting the toxicity or antigenic of JEV. Conclusion Two new JEV isolates were both belonging to genotype Ⅰ, and the key sites of amino acid were not changed.
