CAJ | 학술논문

目的了解浙江省部分地区蚊媒携带流行性乙型脑炎(乙脑)病毒(JEV)情况及其分子流行病学特征.方法 2009-2010年7-8月从浙江省慈溪市和仙居县乙脑监测点采集蚊13620只,利用荧光定量PCR方法检测蚊中JEV核酸,用BHK-21细胞分离病毒,扩增JEV分离株E基因,然后选择6株进行测序并进行同源性分析.结果慈溪和仙居监测点蚊中JEV带病毒率分别为17.0％(27/159)和3.4％( 1/29);从2010年143批蚊中分离到22株JEV,分离率为15.4％.6株测序株E基因全长均为1500个核苷酸(nt),推导编码500个氨基酸(aa),未发现nt插入和丢失,nt和aa同源性分别为99.2％～ 99.8％和100.0％,与浙江省2007-2008年JEV分离株的nt和aa同源性分别为99.1％～99.3％和99.2％～ 99.8％,与乙脑疫苗株SA14-14-2同源性分别为87.6％～88.0％和97.8％.E基因进化树显示,6株测序株位于基因Ⅰ型分支上.结论浙江省慈溪市和仙居县监测点蚊媒携带JEV,其中慈溪监测点蚊媒中乙脑带病毒率明显高于仙居监测点,JEV分离株均为基因Ⅰ型.
목적 료해절강성부분지구문매휴대류행성을형뇌염(을뇌)병독(JEV)정황급기분자류행병학특정.방법 2009-2010년7-8월종절강성자계시화선거현을뇌감측점채집문13620지,이용형광정량PCR방법검측문중JEV핵산,용BHK-21세포분리병독,확증JEV분리주E기인,연후선택6주진행측서병진행동원성분석.결과 자계화선거감측점문중JEV대병독솔분별위17.0％(27/159)화3.4％( 1/29);종2010년143비문중분리도22주JEV,분리솔위15.4％.6주측서주E기인전장균위1500개핵감산(nt),추도편마500개안기산(aa),미발현nt삽입화주실,nt화aa동원성분별위99.2％～ 99.8％화100.0％,여절강성2007-2008년JEV분리주적nt화aa동원성분별위99.1％～99.3％화99.2％～ 99.8％,여을뇌역묘주SA14-14-2동원성분별위87.6％～88.0％화97.8％.E기인진화수현시,6주측서주위우기인Ⅰ형분지상.결론 절강성자계시화선거현감측점문매휴대JEV,기중자계감측점문매중을뇌대병독솔명현고우선거감측점,JEV분리주균위기인Ⅰ형.
Objective To investigate the molecular characteristics of Japanese encephalitis virus (JEV) living in vector mosquitoes,from Zhejiang province.Methods A total of 13620 mosquitoes were collected from the monitoring stations located in Cixi city and Xianju county in Zhejiang province,in July and August,2009-2010.Nucleic acid of JEV from the mosquitoes was monitored by using real-time RT-PCR.The virus strains were isolated with BHK-21 cell line,with E genes of the isolated viruses amplified,sequenced and their phylogeny and homology analyzed.Results The positive rates of JEV for those mosquitoes collected in the stations of Cixi and Xianju were 17.0％ (27/159) and 3.4％ ( 1/29 ),respectively.Twenty-two JEV strains were isolated,accounted for 15.4％ among the 143 batches of mosquitoes collected in 2010.All E genes in the 6 sequenced virus isolates contained 1500 nucleotides encoding 500 amino acids,in which no inserts and deletions were identified.The identity rates of nucleotide and amino acid in E gene were 99.2％-99.8％ and 100.0％ among the 6 JEV strains isolated from Zhejiang,99.1％-99.3％ and 99.2％-99.8％ between the Zhejiang strains in 2009-2010 and the Zhejiang strains in 2007-2008,respectively,87.6％-88.0％ and 97.8％ between the 6 Zhejiang strains and the vaccine strain SA 14-14-2 of JEV,respectively.The phylogeny tree of E gene indicated that the JEV isolates in Zhejiang during 2009-2010 was located in the branch of the genotype Ⅰ.Conclusion Mosquitoes collected from Cixi and Xianju areas carried JEV,with the rate of JEV in Cixi higher than in Xianju.All the Zhejiang isolates in 2009-2010 were proven to be the genotype Ⅰ of JEV.