中国乙脑病毒病原学特征课件.ppt

1、中国乙脑病毒的病原学特征,王环宇中国疾病预防控制中心 病毒病预防控制所 病毒性脑炎室,乙脑病原学背景,1949年，在中国北京分离到乙脑病毒。黄祯祥先生 黄祯祥，王逸民. 北京市流行性脑炎病毒的分离及鉴别. 中华医学杂志，1951，37（4）：280-285.1970年代，在中国使用乙脑灭活疫苗株（P3）。1982年，马中使用的疫苗株（SA14-2-8）。陈伯权先生 Bo Quan Chen and Barry J. Beaty. Japanese encephalitis vaccine (2-8 strain) and parent (SA14 strain) Virus in Culex

2、tritaeniorhynchus mosquitoes. Am. J. Trop. Med. Hyp., 31(2), 1982, 403-407.1989年，人中使用的减毒活疫苗株 （SA14-14-2）。余永新先生从2001-2006，在全国18个省开展乙脑病原学监测工作，共分离并鉴定到 100余株乙脑病毒。分子流行病学分析显示，目前中国乙脑病毒为基因1型和基因3型共同流行。 Huan Yu Wang, Tomohiko Takasaki, Shi Hong Fu, et al. Molecular epidemiological analysis of Japanese Encepha

3、litis virus (JEV) in China. J Gen Virol., 88, 2007, 885-894.,标本采集广西省（20052006）,north latitude : 24302930 Altitude: 670m7640m,Yunnan Province,Northwest of Yunnan,China,标本分拣、运输及保存,液氮,病毒的细胞病变和动物试验BHK-21 细胞72小时出现细胞病变圆缩、脱落C6/36细胞72小时出现病变时间圆缩、聚集、融和乳鼠3-4天，发病死亡,细胞病变和动物试验,ELISA检测,IFA检测 ( 40),采用 3 种抗体用ELISA方法

4、乙脑病毒多克隆抗体甲病毒多克隆抗体布尼亚病毒多克隆抗体病毒与乙脑抗体反应病毒与甲病毒组特异抗体反应病毒与以上 3 种抗体均不反应,血清学检测,乙脑病毒基因组结构,目的基因的扩增测序,中国乙脑病毒进化分析E基因,不同基因型别中国乙脑病毒的分布,E基因核苷酸及氨基酸差异分析,乙脑病毒PrM基因区段氨基酸差异分析,G3型乙脑病毒流行株与减毒活疫苗株PrM区段氨基酸差异,G1型乙脑病毒流行株与减毒活疫苗株PrM区段氨基酸差异,基因3型乙脑病毒E基因区段氨基酸差异分析,E基因乙脑病毒的毒力中国乙脑野毒株与SA14-14-2株在E基因存在8个氨基酸位点差异G3型乙脑野毒株与SA14-14-2株在E基因存在

5、3个氨基酸位点差异,核苷酸CTT GAA ACC GAG CAG AAG GCG GGTATG TCT TC (T) C,G1型乙脑病毒流行株与减毒活疫苗株E区段氨基酸差异,氨基酸 F(Phe)苯病氨酸 K(Lys)赖氨酸 A(Ala)丙氨酸 G(Gly)苷氨酸 H(His)组氨酸 M(Met)甲硫氨酸 V(Val)缬氨酸 D(Asp)天冬氨酸T(Thr)苏氨酸 A (Ala)丙氨酸 A (Ala)丙氨酸,氨基酸L(Leu)亮氨酸 E(Glu)谷氨酸 T(Thr)苏氨酸 E(Glu)谷氨酸 Q(Gln)谷氨酰胺 K(Lys)赖氨酸 A(Ala)丙氨酸 G(Gly)苷氨酸M(Met)甲硫氨酸 S

6、(Ser)丝氨酸 S (Ser)丝氨酸,氨基酸位点E107E138E177E244E264E279E315E447E129E222E366,核苷酸TTT AAA GCC GGG CAT ATG GTG GATACA GCT GCC,SA14-14-2,G1 型 JEV,中国乙脑野毒株与SA14-14-2株在E基因存在8个氨基酸位点差异G1型乙脑野毒株与SA14-14-2株在E基因存在3个氨基酸位点差异,基因1型乙脑病毒E基因区段氨基酸差异分析,SH-53/Shanghai/2001 :TTTAGACAG-GATTAAG-TCATGTGT-GTAATGTGAG-ATAAGAAAATGTGCATG

7、TGGAGTCAGGCCAG SH-81/Shanghai/2001 :TTTAGACAG-GATTAAG-TCATGTG-AG-ATAAGAAAATG-TGCATGTGGAGTCAGGCCAG SH-83/Shanghai/2001 :TTTAGACAG-GATTAAG-TCATGTGT-GTAATGTGAG-ATGAGAAAATGTGCATGTGGAGTCAGGCCAG SH03-103/Shanghai/2003 :TTTAGACAG-GATTAAG-TCATGTGT-GTAATGTGAG-ATAAGAAAATG-TGCATGTGGAGTCAGGCCAG SH03-127/Shangha

8、i/2003 :TTTAGACAG-GATTAAG-TCATGTG-AG-ATAAGAAAATG-TGCATGTGGAGTCAGGCCAG SH03-129/Shanghai/2003 :TTTAGACAG-GATTAAG-TCATGTG-AG-ATAAGAAAATG-TGCATGTGGAGTCAGGCCAG SH05-24/Shanghai/2005 :TTTAGACAG-GATTAAG-TCATGTG-AG-ATAAGAAAATGTGCATGTGGAGTCAGGCCAGMo/Ishikawa/1994 :TTTAGACAG-GATTAAG-TCATGTGT-GTAATGTGAG-ATAAG

9、AAAATG-TGCATGTGGAGTCAGGCCAG Sw/Tokyo/1/1994 :TTTAGACAG-GATTAAG-TCATGTGT-GTAATGTGAG-ATAAGAAAATG-TGCATGTGGAGTCAGGCCAG Sw/Chiba/88/2002 :TTTAGACAG-GGTTAAG-TCATGTGT-GTAATGTGAG-ATAAGAAAATG-TGCATGTGGAGTCAGGCCAG Sw/Hiroshima/25/2002 :TTTAGACAG-GATTAAG-TCATGTGT-GTAATGTGAG-ATAAGAAAATG-TGCATGTGGAGTCAGGCCAG Sw

10、/Kagawa/27/2002 :TTTAGACAG-GATTAAG-TCATGTGT-GTAATGTGAG-ATAAGAAAATG-TGCATGTGGAGTCAGGCCAG Sw/Mie/41/2002 :TTTAGACAG-GATTAAG-TCATGTGT-GTAATGTGAG-ATAAGAAAATG-TGCATGTGGAGTCAGGCCAG Sw/Shizuoka/33/2002 :TTTAGACAG-GATTAAG-TCATGTGT-GTAATGTGAG-ATAAGAAAATG-CGCATGTGGAGTCAGGCCAG Sw/Okinawa/285/2003 :TCTAGACAG-GA

11、TTAAG-TCATGTGT-GTAATGTGAG-ATAAGAAAATG-TGCATGTGGAGTCAGGCCAG Sw/Kagawa/35/2004 :TTTAGACAG-GATTAAG-TCATGTG-AG-ATAAGAAAATG-TGCATGTGGAGTCAGGCCAG Sw/Mie/34/2004 :TTTAGACAG-GATTAAG-TTATGTGT-GTAATGTGAG-ATAAGAAAATG-TGCATGTGGAGTCAGGCCAG Sw/Mie/40/2004 :TTTAGACAG-GATTAAG-TCATGTGT-GTAATGTGAG-ATAAGAAAATG-TGCATGT

12、GGAGTCAGGCCAGA2/BJ/1949 :TCTAGTGTG-ATTTAAGGCAGAAAAATAAATTATGTAA-ATAATGTAA-ATGAGAAAATGTATGTATACGGAGTCAGGCCAG Beijing/1/1949 :TCTAGTGTG-ATTTAAGGCAGAAAAATAAATTATGTAA-ATAATGTAAATGAGAAAATGTATGTATACGGAGTCAGGCCAG China/SA14/1954 :TCTAGTGTG-ATTTAAGGTAGAAAAGTAGACTATGTAA-ATAATGTAA-ATGAGAAAATGCATGCATATGGAGTCAG

13、GCCAG K82P01/Korea/1982 :TCTAGTGTG-ACTTAAGGTAGAAATGTAAATAATGTAA-A-TG-AGAAAATGCATGCATATGGAGTCAGGCCAG K87P39/Korea/1987 :TCTAGTGTG-ACTTAAGGTAGAAATGTAAATAATGTGA-A-TG-AGAAAATGCATGTATATGGAGTCAGGCCAG 47/China/1950s :TCTAGTGTG-ACTTAAGGTAGAAATGTAAATAATGTAA-A-TG-AGAAAATGCATGCATATGGAGTCAGGCCAGFU/1995 :TTTAGAT

14、AGCAAATCAAG-TTAAGTGT-ATAATGTGA-ATAAGAAAATG-TACATATGGAGTCAGGCCAGIndonesia/JKT6468/19 :TCTAGTGTG-GTCCCAAG-TAATAAAATGAATGTAACAAAATGAATGTATAATATAGGGTGTACATATGGAGTCAGGCCAG,乙脑病毒的3NTR序列分析,中国乙脑病毒 3NTR 基因区段基因缺失分析,终止 密码子,3,1,3 NTR,可变区域,2,4,基因 型别,注：上三角部分为核苷酸同源性结果，下三角部分为氨基酸同源性结果,毒株名称02-76 Beijing-1 P3 SA-14 SA1

15、4-14-2 SA14-12-1-7 JaGAr01 JaOH0566 ML17 JaOArS982 JEV-AT31 K87P39 Ling RP-2ms RP-9 CH1392 CH2195LA CH2195SA T1P1 HVI TC TL YL P20778 GP78 SH-53 Ishikawa K94P05 KV1899 FU JKT6468,297.798.9 99.2 98.4 98.6 99.2 99.1 98.9 99.2 99.2 99.2 99.4 98.9 99.0 99.0 98.5 98.5 99.0 98.8 98.6 98.7 98.4 98.6 98.3 9

16、8.097.5 96.7 97.0 97.6 95.0,398.7 97.499.4 98.6 98.8 99.4 99.3 99.1 99.2 99.4 99.2 98.7 99.1 99.2 99.3 98.6 98.7 99.3 99.0 98.6 98.7 98.7 98.6 98.5 98.0 97.5 96.9 97.0 97.6 95.0,499.4 97.6 98.799.0 99.2 99.7 99.6 99.4 99.6 99.7 99.6 99.0 99.4 99.5 99.5 98.9 99.0 99.5 99.3 99.0 99.2 98.9 99.0 98.7 98

17、.3 97.8 97.1 97.3 97.9 95.3,598.9 97.2 98.3 99.499.2 99.0 98.9 98.7 98.8 99.0 98.8 98.2 98.7 98.8 98.8 98.1 98.2 98.8 98.5 98.2 98.4 98.3 98.2 98.0 97.6 97.0 96.4 96.6 97.1 94.6,699.0 97.3 98.3 99.5 99.699.2 99.1 98.9 99.0 99.2 99.1 98.5 95.9 99.0 99.0 98.4 98.5 99.0 98.7 98.4 98.6 98.5 98.5 98.3 97

18、.9 97.3 96.6 96.9 97.4 94.8,798.8 97.4 98.3 98.8 98.3 98.499.7 99.5 99.7 99.9 99.7 99.1 99.6 99.7 99.7 99.0 99.1 99.7 99.4 99.1 99.1 99.1 99.1 98.8 98.4 97.8 97.2 97.4 98.0 95.4,898.1 96.9 97.7 98.1 97.7 97.8 98.999.7 99.5 99.7 99.5 98.9 99.4 99.5 99.5 98.8 98.9 99.5 99.2 99.0 99.0 99.0 98.9 98.7 98

19、.3 97.7 97.1 97.3 97.9 95.4,998.0 96.8 97.6 98.0 97.6 97.7 98.9 99.899.3 99.5 99.3 98.7 99.2 99.3 99.4 98.7 98.7 99.4 99.1 98.8 98.8 98.8 98.7 98.5 98.1 97.6 96.9 97.1 97.7 95.2,1098.9 97.0 97.8 98.6 98.1 98.2 98.0 97.4 97.399.6 99.5 99.0 99.3 99.4 99.4 99.1 99.2 99.4 99.2 98.8 99.0 98.8 99.0 98.7 9

20、8.3 97.7 97.3 97.3 97.9 95.3,1198.8 97.4 98.2 98.7 98.3 98.4 99.8 98.9 98.8 97.999.7 99.0 99.6 99.7 99.7 99.0 99.1 99.7 99.4 99.0 99.1 99.1 99.0 98.8 98.4 97.8 97.1 97.3 98.0 95.4,1297.8 96.3 97.0 97.8 97.3 97.4 97.3 96.7 96.6 97.0 97.399.0 99.3 99.4 99.5 98.9 99.0 99.5 99.2 98.9 99.0 98.9 99.0 98.7

21、 98.5 97.9 97.1 97.3 98.0 95.5,1397.5 98.9 97.1 97.4 96.9 97.0 97.2 96.6 96.5 96.9 97.1 96.198.7 98.8 98.9 98.3 98.4 98.9 98.6 98.4 98.5 98.3 98.5 98.2 97.8 97.3 96.6 96.8 97.4 94.9,1498.5 97.2 98.0 98.5 98.0 98.1 99.4 98.6 98.6 97.7 99.6 97.0 96.999.7 99.4 98.7 98.7 99.4 99.1 98.7 98.8 98.8 98.7 98

22、.5 98.1 97.5 96.8 97.0 97.7 95.1,1598.5 97.2 98.0 98.5 98.1 98.1 99.5 98.7 98.6 97.8 99.6 97.0 97.0 99.999.5 98.8 98.9 99.5 99.2 98.8 98.9 98.9 98.8 98.6 98.2 97.6 96.9 97.1 97.8 95.2,1698.7 97.3 98.2 98.6 98.2 98.3 99.7 98.9 98.8 97.8 99.8 97.2 97.1 99.5 99.698.8 98.9 99.9 99.2 98.9 98.9 99.0 98.9

23、98.6 98.3 97.7 97.0 97.2 97.8 95.3,1797.2 95.6 96.3 97.1 96.6 96.7 96.5 96.0 95.9 97.3 96.4 95.7 95.4 96.2 96.3 96.399.7 98.8 98.5 98.2 98.3 98.2 98.4 98.1 97.7 97.1 96.6 96.7 97.3 94.9,1897.2 95.6 96.3 97.1 96.6 96.7 96.5 96.0 95.9 97.3 96.4 95.7 95.5 96.2 96.3 96.4 99.998.9 98.6 98.3 98.4 98.3 98.

24、4 98.2 97.8 97.2 96.6 96.7 97.4 95.0,1998.7 97.3 98.1 98.6 98.2 98.3 99.7 98.8 98.8 97.8 99.8 97.1 97.1 99.5 99.5 99.9 96.3 96.399.2 98.9 98.9 99.0 98.9 98.6 98.3 97.7 97.0 97.3 97.8 95.3,2098.5 97.2 98.1 98.5 98.1 98.2 99.1 98.5 98.4 97.7 99.1 97.0 97.0 98.8 98.8 99.0 96.2 96.2 99.098.7 98.8 98.7 9

25、8.6 98.4 98.0 97.4 96.7 96.9 97.6 95.0,2198.1 97.1 97.6 98.2 97.7 97.8 98.4 97.8 97.7 97.3 98.3 96.7 96.8 98.1 98.1 98.3 95.8 95.8 98.2 98.598.7 98.3 98.3 98.0 97.7 97.1 96.4 96.6 97.2 94.7,2297.9 96.9 97.5 98.1 97.6 97.7 98.4 97.9 97.8 97.2 98.4 96.5 96.6 98.1 98.2 98.3 95.8 95.8 98.3 98.4 99.098.3

26、 98.4 98.1 97.8 97.2 96.5 96.7 97.3 94.7,2398.4 97.1 97.9 98.4 98.0 98.0 99.4 98.6 98.5 97.6 99.6 96.9 96.9 99.3 99.3 99.5 96.1 96.1 99.5 98.8 98.0 98.098.3 98.0 97.7 97.1 96.5 96.7 97.2 94.7,2497.1 96.7 96.8 97.1 96.6 96.7 96.9 96.3 96.2 96.4 96.9 95.9 96.4 96.6 96.7 96.8 95.1 95.1 96.8 96.6 96.3 9

27、6.1 96.598.2 98.0 97.4 96.7 96.9 97.5 95.0,2597.7 96.1 97.1 97.8 97.3 97.4 97.1 96.6 96.5 97.2 97.1 96.1 95.9 96.9 96.9 97.0 95.7 95.7 97.0 96.9 96.6 96.5 96.8 95.697.6 97.0 96.4 96.6 97.1 94.7,2789.1 88.8 89.0 89.1 88.7 88.8 89.0 88.6 88.5 88.7 89.0 88.7 88.5 88.8 88.8 88.9 88.2 88.2 88.9 88.8 88.7

28、 88.6 88.7 88.7 88.6 98.597.7 97.9 97.6 94.4,2889.5 88.9 89.3 89.4 88.9 89.1 89.3 88.9 88.9 89.3 89.3 88.9 88.7 89.0 89.1 89.2 88.5 88.5 89.2 89.1 88.9 88.9 89.0 88.8 88.9 97.5 97.097.2 96.8 93.8,2988.9 88.5 88.7 88.8 88.4 88.5 88.8 88.4 88.3 88.5 88.8 88.4 88.3 88.6 88.6 88.7 87.9 87.9 88.7 88.6 88

29、.4 88.4 88.5 88.5 88.3 98.1 97.6 96.996.9 93.9,3089.6 89.2 89.3 89.4 89.0 89.1 89.5 89.0 89.0 89.3 89.4 89.1 88.7 89.2 89.3 89.3 88.8 88.9 89.3 89.3 89.0 89.0 89.1 89.0 88.6 90.2 90.0 89.6 89.794.6,3184.9 84.6 84.8 84.9 84.6 84.6 85.0 84.8 84.7 84.8 84.9 84.7 84.5 84.8 84.8 84.8 84.5 84.5 84.9 84.7

30、84.7 84.6 84.7 84.6 84.3 83.5 83.4 83.0 83.2 83.7,199.2 99.4 99.7 99.0 99.2 99.8 99.6 99.4 99.7 99.8 99.7 99.1 99.4 99.6 99.6 99.0 99.1 99.6 99.3 99.0 99.1 99.0 99.1 98.8 98.597.9 97.2 97.4 98.0 95.4,毒株编号,No.1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31,2689.2 8

31、8.8 89.1 89.2 88.7 88.9 89.1 88.7 88.7 88.9 89.1 88.8 88.6 88.9 89.0 89.0 88.2 88.3 89.0 88.9 88.8 88.7 88.8 88.8 88.799.3 98.2 98.4 98.1 95.0,02-76株、SH-53株与其他乙脑病毒全基因组核苷酸及氨基酸同源性比较,全基因序列核苷酸及氨基酸差异分析,全基因序列的系统进化分析,分离自不同国家的31株乙型脑炎病毒全基因序列系统进化分析， MVE作为外群。A: PrM/C区段，B: 3NTR区段，C: E区段，D: 全基因。,谢 谢 大 家,王环宇 中国疾病预防控制中心病毒病预防控制所 病毒性脑炎实验室 电话：010-83559924 手机：13621199546 E-mail：,