中国人兽共患病学报
中國人獸共患病學報
중국인수공환병학보
CHINESE JOURNAL OF ZOONOSES
2012年
7期
695-699
,共5页
顾雯%金聪%张硕%张全福%李建东%杭小同%王芹%李川%梁米芳%李德新
顧雯%金聰%張碩%張全福%李建東%杭小同%王芹%李川%樑米芳%李德新
고문%금총%장석%장전복%리건동%항소동%왕근%리천%량미방%리덕신
登革3型病毒%义鸟分离株%生物学特性
登革3型病毒%義鳥分離株%生物學特性
등혁3형병독%의조분리주%생물학특성
dengue virus type 3%Yiwu strain%biological characteristics
目的 对2009年浙江省义鸟市暴发流行的登革3型病毒进行系统进化分析并初步研究其E蛋白的免疫原性.方法 RT-PCR扩增义乌分离株全基因组,利用MEGA 4分别构建E及NS1区域核苷酸及氨基酸系统发生树,进行系统进化分析.构建重组质粒JESS -ywD3 prM/E397JEV-pcDNA5,IFA及Western Blot检测E蛋白在293T细胞中的表达及分泌情况.将纯化后的分泌性E蛋白免疫8ALB/c小鼠,通过ELISA、中和试验等方法对体液免疫进行测定.结果 登革3型病毒义乌分离株与广州GZID3株及印度GWIL-25株核苷酸及氨基欧序列同源性均达到99%以上；其分泌性E蛋白可以刺激小鼠产生登革特异性IgG抗体及针对同型病毒的中和抗体.结论 义乌分离株属于登革3型病毒亚型Ⅲ,其分泌性E蛋白具有一定的免疫原性.
目的 對2009年浙江省義鳥市暴髮流行的登革3型病毒進行繫統進化分析併初步研究其E蛋白的免疫原性.方法 RT-PCR擴增義烏分離株全基因組,利用MEGA 4分彆構建E及NS1區域覈苷痠及氨基痠繫統髮生樹,進行繫統進化分析.構建重組質粒JESS -ywD3 prM/E397JEV-pcDNA5,IFA及Western Blot檢測E蛋白在293T細胞中的錶達及分泌情況.將純化後的分泌性E蛋白免疫8ALB/c小鼠,通過ELISA、中和試驗等方法對體液免疫進行測定.結果 登革3型病毒義烏分離株與廣州GZID3株及印度GWIL-25株覈苷痠及氨基歐序列同源性均達到99%以上；其分泌性E蛋白可以刺激小鼠產生登革特異性IgG抗體及針對同型病毒的中和抗體.結論 義烏分離株屬于登革3型病毒亞型Ⅲ,其分泌性E蛋白具有一定的免疫原性.
목적 대2009년절강성의조시폭발류행적등혁3형병독진행계통진화분석병초보연구기E단백적면역원성.방법 RT-PCR확증의오분리주전기인조,이용MEGA 4분별구건E급NS1구역핵감산급안기산계통발생수,진행계통진화분석.구건중조질립JESS -ywD3 prM/E397JEV-pcDNA5,IFA급Western Blot검측E단백재293T세포중적표체급분비정황.장순화후적분비성E단백면역8ALB/c소서,통과ELISA、중화시험등방법대체액면역진행측정.결과 등혁3형병독의오분리주여엄주GZID3주급인도GWIL-25주핵감산급안기구서렬동원성균체도99%이상；기분비성E단백가이자격소서산생등혁특이성IgG항체급침대동형병독적중화항체.결론 의오분리주속우등혁3형병독아형Ⅲ,기분비성E단백구유일정적면역원성.
This study performed phylogenic analysis on a dengue strain isolated from an outbreak of dengue fever in 2009 at Yiwu City of Zhejiang Province,China,and further to analyze the immunogenicity of E protein of this viral isolate.Firstly,the viral genome was amplified by RT-PCR and phylogenetic trees were constructed by MEGA 4 based on both nucleotide and amino acid sequences of E and NS1 proteins.The phylogenetic analysis showed that the similarity of Yiwu strain with the Guangzhou GZ1D3 strain and the India GWL-25 strain was over 99％.Secondly,the expression plasmid of E protein was constructed and transfected into 293T cells.The secreted E protein were then purified by sucrose density gradient centrifugation and used to inoculate BALB/c mice.The humoral immunity was evaluated by ELISA and neutralizing antibody analysis.Resuits showed that the E protein of Yiwu strain could induce dengue specific IgG antibodies and neutralizing antibodies.Therefore,the study found that the Yiwu strain was classified into the subtype Ⅲ of dengue virus type 3 (DENV-3),and the E protein of this strain had strong immunogenicity
