下载：人感染H7N9禽流感病毒病原学特点及实验室检测
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Transcript 下载：人感染H7N9禽流感病毒病原学特点及实验室检测
人感染H7N9禽流感病毒
病原学特点及实验室检测
王 大 燕
世界卫生组织流感参比与研究合作中心(WHO CC)
国家流感中心(CNIC) 病毒病预防控制所(IVDC)
中国疾病预防控制中心 (China CDC)
2013年4月1日
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
病例1
病例2
病例3
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
既往禽H7Nx病毒简介
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
HPAI H7N3
LPAI H7N9
HPAI H7N7 、
H7N3、 H7N1
LPAI H7Nx、H7N2、
H7N3、H7N1、H7N9
LPAI H7N9
LPAI H7N8、
H7N9（Korea）
HPAI H7N3
（Pakistan）
HPAI H7N3
LPAI H7N9
HPAI
H7N7、
H7N3 、
H7N4
禽中暴发的H7
2006年前暴发的均为HPAI H7
欧洲：意大利、英国、德国、比利时、荷兰、丹麦、挪威、 捷克、西班牙、瑞典
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
近年来人感染禽流感病毒病例
病毒亚型
病原来源
时间/国家
病例数/临床表现
A/H7N7
鸥
1980/美国
3/结膜炎
A/H7N7
鸭
1996/英国
1/结膜炎
A/H5N1
禽
1997/香港
18/ILI，肺炎
A/H9N2
鸡
1998/中国广东
5/ ILI，肺炎
A/H9N2
禽
1999/香港
2/ILI
A/H7N2
禽
2002/北美（Virginia） 1/ILI，血清学诊断
A/H9N2
禽
A/H5N1
禽
2003/香港
1/ILI
血清学证据：
2003/香港
2/ ILI，肺炎
禽
89/结膜炎、ILI、肺炎; 1名死亡病例
人感染H4,
H5, H6,2003/荷兰
H7,H9,H10,
H11 AIVs.
A/H7N7
A/H5N1
禽
2003至今, 15个国家
602/ ILI, 肺炎
A/H7N2(NY/107)
来源不清
2003/北美(New York)，1/肺炎
A/H7N3
禽
2004/加拿大
2/结膜炎、ILI; 1名LPAI；1名HPAI
A/H5N2
禽
2005/日本(Ibaraki)
13/无临床症状，血清学诊断
A/H7N2
禽
2007/英国(Wales)
1/结膜炎、ILI
A/H9N2
来源不清
2007/香港
1/ ILI
A/H10N7
鸭
2004/埃及( Ismaillia)
2/ ILI
A/H10N7
来源于疫区的鸟
2010/悉尼(New South 7/上呼吸道感染症状、结膜炎
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
Wales)
CDC. Avian Influenza A Virus Infections
of Humans. http://www.cdc.gov/flu/avian/gen-info/avian-flu-humans.htm.
HPAI
89/ Netherlands
H7N7
1/ Canada
1 died
H7N3
4/USA
H7N7
1/USA
H7N7
1959
LPAI
1/USA
H7N7
1996 2002
2003
2006 2007
2004
1977 19791/England
1/ USA
1980
7/
Italy
1/ Canada
H7N7
H7N2
H7N3
H7N3
1/ USA
H7N2
2/ Mexico
H7N3
4/ Wales
H7N2
1/ Norfolk
H7N3
H7感染人（>100人）
美国、英国、意大利、荷兰、加拿大
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
H7N9病毒
亚洲, 7
欧洲, 5
美洲,
13
共25株
均为LPAI
无感染人病例
亚洲：7株
均来自野禽
5株—韩国
2株—蒙古
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
H7N9的致病力和传播力
HPAI H7N3
LPAI H7N9
HPAI H7N3
LPAI H7N9
LPAI H7N9
Nose (white bars) , lung (black bars)
HPAI H7N3和LAPI H7N9均不需经过适应便
高致病性的HPAI H7N3致小鼠死亡，
可在小鼠体内有效复制，且均能在雪貂上
在雪貂模型中可通过接触传播，但
下呼吸道有效复制。
LAPI H7N9在雪貂模型中不传播。
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
中国地区H7 AIV
A/Baer's pochard/HuNan/414/2010(H7N1)
A/chicken/Hebei/1/2002(H7N2)
A/duck/Guangdong/1/1996(H7N3)
A/duck/Hubei/126/1985(H7N8)
A/duck/Jiangxi/1742/03(H7N7)
A/duck/Jiangxi/1760/03(H7N7)
A/duck/Jiangxi/1786/03(H7N7)
A/duck/Jiangxi/1814/03(H7N7)
A/duck/Nanchang/1904/1992(H7N1)
A/duck/Nanchang/1944/92(H7N1)
A/duck/Zhejiang/10/2011(H7N3)
A/duck/Zhejiang/12/2011(H7N3)
A/duck/Zhejiang/2/2011(H7N3)
A/magpie-robin/China/28710/93(H7)
亚型：H7N1, H7N2,
H7N3, H7N7, H7N8
野禽居多
LPAI
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
人感染H7N9禽流感病毒
实验室检测
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
核酸检测
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
病毒分离
从3个病例中分离到3株病毒
• A/Shanghai/1/2013(H7N9)
• A/Shanghai/2/2013(H7N9)
• A/Anhui/1/2013(H7N9)
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
序列测定
PB2
PB1
PA
HA
NP
NA
MP
NS
6个内部片段来源于H9N2
三株H7N9病毒为同一基因类型
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
序列特点
耐药性分析
对烷胺类药物是耐药的，即M2蛋白25-42位氨基酸发生S31N突变。
HA连接肽分析
HA蛋白339位连接肽由单个（PKGRGLFG）碱性氨基酸组成，根据既往
研究，具有这种序列特点的病毒对禽类呈低致病性。
受体结合位点及其它重要位点分析
Shanghai/1/2013的HA蛋白 226-228位氨基酸属于禽源受体结合位点
（为QSG）。Shanghai/2/2013和Anhui/1/2013的HA蛋白 226-228位氨基
酸受体结合位点（为LSG）
PB2蛋白627位氨基酸为K，提示致病性增强。
NS1蛋白92位氨基酸为D，与禽流感病毒一致，而与人流感病毒不同。
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
HA核酸同源性比较
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
A/northern_pintail/Akita/1370/2008_H7N7_Japan
A/mallard/Korea/GG3/2007_H7N7_South_Korea
A/environment/Mongolia/1-51/2007_H7N7_Mongolia
100 A/duck/Korea/BC10/2007_H7N3_South_Korea
A/duck/Mongolia/119/2008_H7N9_Mongolia
A/duck/Korea/A349/2009_H7N2_South_Korea
99
A/duck/Thailand/CU-LM7291T/2010_H7N6_Thailand
89
A/quail/Thailand/CU-J2882/2009_H7N1_Thailand
98
92
A/wild_bird/Korea/5-77/2005_H7N8_South_Korea
A/duck/Zhejiang/12/2011_H7N3_China
A/Shanghai/1/2013
99
100
A/Shanghai/2/2013
100
99 A/Anhui/1/2013
A/duck/Jiangxi/1742/03_H7N7_China
A/wild_bird_feces/Korea/HDR16/2003_H7N2_South_Korea
86
A/duck/Mongolia/867/2002_H7N1_Mongolia
98
AY338455_A/Netherlands/127/03_H7N7_Netherlands
71
100
AY338457_A/Netherlands/33/03_H7N7_Netherlands
AY338459_A/Netherlands/219/2003_H7N7_Netherlands
A/chicken/Italy/444/99_H7N1_Italy
A/chicken/Italy/322/2001_H7N1_Italy
A/quail/Italy/4610/2003_H7N2_Italy
76
97
A/turkey/Italy/5425/2007_H7N3_Italy
A/mallard/Sweden/100993/2008_H7N7_Sweden
98
88
98
A/mute_swan/Hungary/5973/2007_H7N7_Hungary
A/mallard/Italy/497-35/2006_H7_Italy
A/goose/Czech_Republic/1848-K9/2009_H7N9_Czech_Republic
98
A/mallard/Korea/822/2010_H7N7_South_Korea
91
A/duck/Thailand/CU-10507T/2011_H7N4_Thailand
78
A/wild_goose/Dongting/PC0360/2012_H7N7_China
100
A/spot-billed_duck/Korea/447/2011_H7N9_South_Korea
84
A/parrot/England/1174/94_H7N1_United_Kingdom
A/ostrich/South_Africa/5352/92_H7N1_South_Africa
96
AF028020_A/England/268/1996_H7N7_United_Kingdom
78
GU053110_A/England/AV877/1996_H7N7_United_Kingdom
100 A/England/AV877/1996_H7N7_United_Kingdom
A/chicken/Pakistan/16/99/95_H7N3_Pakistan
100 A/chicken/Karachi/NARC-23/2003_H7N3_Pakistan
A/quail/Aichi/1/2009_H7N6_Japan
87
A/duck/Hong_Kong/301/1978_H7N2_Hong_Kong
99
A/black_kite/Egypt/054/2004_H7N7_Egypt
A/chicken/Jena/1816/87_H7N7_Germany
87
89
A/chicken/Hebei/1/2002_H7N2_China
78
100 A/African_starling/England-Q/983/1979_H7N1_United_Kingdom
A/duck/Vietnam/OIE-0178/2012_H7N1_Viet_Nam
A/mallard/New_Zealand/1365-355/2005_H7N7_New_Zealand
94
100 A/equine/Prague/1/1956_H7N7_Czech_Republic
100
A/equine/Cambridge/1/63_H7N7_United_Kingdom
A/equine/Argentina/1/1977_H7N7_Argentina
A/chicken/Chile/184240-1/2002_H7N3_Chile
100
A/cinnamon_teal/Bolivia/4537/2001_H7N3_Bolivia
92
A/turkey/Oregon/1971_H7N3_USA
A/shorebird/Delaware_Bay/274/1994_H7N5_USA
100
A/mallard/Delaware/418/2005_H7N3_USA
100
82
A/blue-winged_teal/Ohio/566/2006_H7N9_USA
100
73
A/green-winged_teal/MD/648/2004_H7N3_USA
CY015006_A/Canada/rv504/2004_H7N3_Canada
A/goose/Nebraska/17097-4/2011_H7N9_USA
CY125728_A/Mexico/InDRE7218/2012_H7N3_Mexico
100
100
100 A/chicken/Jalisco/CPA1/2012_H7N3_Mexico
A/pheasant/Minnesota/917/1980_H7N3_USA
99
88
A/green-winged_teal/ALB/228/1985_H7N3_Canada
A/laughing_gull/Delaware/22/2002_H7N3_USA
A/duck/Guangdong/1/1996_H7N3_China
81
A/turkey/New_York/Sg-00255/1994_H7N2_USA
A/quail/PA/20304/1998_H7N2_USA
A/Guinea_fowl/New_York/10196-7/2005_H7N2_USA
A/chicken/New_York/63806-12/2005_H7N2_USA
78
100
A/turkey/VA/66/02_H7N2_USA
99
EU587368_A/New_York/107/2003_H7N2_USA
A/Guinea_fowl/New_York/31621-8/2005_H7N2_USA
A/chicken/Victoria/1976_H7N7_Australia
A/duck/Tasmania/277/2007_H7N2_Australia
100
100
A/duck/Victoria/512/2007_H7N6_Australia
100
A/chicken/Queensland/1994_H7N3_Australia
100
A/chicken/New_South_Wales/2/1997_H7N4_Australia
91
A/fowl/Egypt/45_H7N1_Egypt
A/chicken/Japan/1925_H7N7_Japan
100 A/duck/Taiwan/33/1993_H7N7_Taiwan
A/chicken/Rostock/45/1934_H7N1_Germany
A/FPV/Dutch/1927_H7N7_United_Kingdom
100 A/chicken/Brescia/1902_H7N7_Italy
100
93
70
H7进化树
红色标记为本次病例
●H7 人感染病例
Eurasian
Equine
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
American
A/mallard/Minnesota/Sg-00177/2007__USA_H12N9
A/mallard/Ohio/2033/2009__USA_H4N9
92
A/American_black_duck/New_Brunswick/03451/2009__Canada_H3N9
A/mallard/Alberta/64/2000__Canada_H11N9
A/mallard/MN/158/2000__USA_H3N9
75
A/northern_shoveler/California/8675/2008__USA_H11N9
A/ruddy_turnstone/Delaware_Bay/220/1995__USA_H7N9
A/mallard/Alberta/193/2000__Canada_H11N9
A/black_duck/Ohio/161/1999__USA_H11N9
89
A/ruddy_turnstone/DE/1538/2000__USA_H7N9
A/American_black_duck/New_Brunswick/02485/2007__Canada_H11N9
A/American_white_pelican/Minnesota/Sg-0611/2008__USA_H13N9
100
A/shorebird/Delaware_Bay/424/2007__USA_H13N9
100
A/shorebird/Delaware_Bay/48/2002__USA_H11N9
80
A/ruddy_turnstone/New_Jersey/980/2002__USA_H2N9
100 A/ruddy_turnstone/Delaware_Bay/96/2002__USA_H1N9
A/thick-billed_murre/Alaska/44085-090/2006__USA_H11N9
100
A/glaucous_gull/Alaska/44199-104/2006__USA_H13N9
99
A/guinea_fowl/Nebraska/17096-1/2011__USA_H7N9
100
89
A/northern_shoverl/Mississippi/11OS145/2011__USA_H7N9
A/king_eider/Alaska/44397-858/2008__USA_H10N9
92
A/blue-winged_teal/Guatemala/CIP049-01/2008__Guatemala_H7N9
81
A/turkey/Minnesota/1/1988__USA_H7N9
A/gull/Maryland/18/1977__USA_H2N9
A/pintail/Alberta/293/1977__Canada_H2N9
96
73
A/Black_Duck/Ohio/239/1986__USA_H11N9
A/ruddy_turnstone/Delaware/2368/1988__USA_H4N9
A/redhead_duck/Alberta/357/1983__Canada_H11N9
83
A/turkey/Wisconsin/1/1968__USA_H5N9
A/tern/Australia/G70C/1975__Australia_H11N9
A/duck/Vietnam/OIE-2386/2009__Viet_Nam_H11N9
100
A/duck/Vietnam/OIE-3301/2011__Viet_Nam_H6N9
A/goose/Zambia/09/2009__Zambia_H11N9
A/mallard/New_Zealand/1440-365/2005__New_Zealand_H11N9
99
A/mallard/Czech_Republic/15902-18K/2009__Czech_Republic_H6N9
93
100
97
A/goose/Czech_Republic/1848-K9/2009__Czech_Republic_H7N9
A/mallard/Switzerland/WV1071028/2007__Switzerland_H11N9
A/Duck/Nanchang/2-0492/2000__China_H2N9(2)
77
A/mallard/Sweden/48/2002__Sweden_H11N9
A/gadwall/Mongolia/OIE-191/2012__Mongolia_H2N9
75
A/duck/Hunan/1590/2007__China_H6N9
A/duck/Mongolia/119/2008__Mongolia_H7N9
A/mallard/Czech_Republic/13438-29K/2010__Czech_Republic_H11N9
A/Anas_crecca/Spain/1460/2008__Spain_H7N9
90
A/duck/Vietnam/LBM81/2012__Viet_Nam_H11N9
99
A/wild_duck/Korea/SH20-27/2008__South_Korea_H7N9
A/shorebird/Korea/S8/2006__South_Korea_H11N9
A/duck/Chiba/7/2006__Japan_H11N9
A/duck/Hokkaido/W245/2004__Japan_H11N9
A/shorebird/Korea/S331/2006__South_Korea_H10N9
A/wild_bird/Korea/A3/2011__South_Korea_H7N9
99
A/wild_bird/Korea/A14/2011__South_Korea_H7N9
A/Shanghai/1/2013
86
A/Shanghai/2/2013
100
98 A/Anhui/1/2013
A/kelp_gull/Argentina/LDC4/2006__Argentina_H13N9
A/emu/NY/12716/1994__USA_H5N9
A/shoveler/Netherlands/19/1999__Netherlands_H11N9
100
A/chicken/Italy/22A/1998__Italy_H5N9
100
100 A/wedge-tailed_shearwater/Western_Australia/2327/1983__Australia_H15N9
A/shearWater/Australia/2576/1979__Australia_H15N9
A/duck/Hong_Kong/278/1978__Hong_Kong_H2N9
A/duck/Hong_Kong/562/1979__Hong_Kong_H10N9
100
98
99
N9 进化树
红色标记为本次病例
American
100
Eurasian
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
0.02
A/Shanghai/1/2013
71
91
A/Shanghai/2/2013
89
A/Anhui/1/2013
A/brambling/Beijing/16/2012_H9N2
A/chicken/Shanghai/C1/2012_H9N2
A/chicken/Zhejiang/329/2011_H9N2
100
A/chicken/Shandong/02/2010_H9N2
A/chicken/Shandong/H/2009_H9N2
A/equine/Guangxi/3/2011_H9N2
99
A/chicken/Zhejiang/607/2011_H9N2
A/chicken/Anhui/HF/2010_H9N2
A/chicken/Jiangsu/Q3/2010_H9N2
82
A/chicken/Shuanggou/1/2011_H9N2
100
A/duck/Shandong/009/2008_China_H5N1
100
93 A/avian/Japan/8KI0162/2008_Japan_H3N8
99
A/avian/Japan/8KI0068/2008_Japan_H3N6
A/environment/Korea/CSM05/2004_South_Korea_H3N1
A/mallard/Jiangxi/8341/2004_China_H6N5
100
90
A/environment/Korea/ESD18/2004_South_Korea_H3N6
A/duck/Guangxi/3333/2006_China_H6N1
A/duck/Guangxi/1248/2006_China_H6N2
99
100
A/wild_bird/Korea/YJ117/2007_South_Korea_H4N6
A/turkey/Italy/977/1999_Italy_H7N1
100 A/turkey/Italy/1067/99__Italy_H7N1
A/turkey/Italy/1265/1999_Italy_H7N1
A/mallard/Czech_Republic/13438-29K/2010_H11N9
88
A/avian/Egypt/920431/2006_H9N2
A/spot-billed_duck/Korea/447/2011_H7N9
81
A/wild_bird/Korea/A14/2011_H7N9
100
A/wild_bird/Korea/A3/2011_H7N9
74
100 A/goose/Czech_Republic/1848-K9/2009_H7N9
A/goose/Czech_Republic/1848-T14/2009_H7N9
PC0360_H7N7-PB2.seq
98
A/duck/Jiangxi/1814/03_China_H7N7
A/wild_duck/Mongolia/1-241/2008_H7N9
A/Anas_crecca/Spain/1460/2008_H7N9
A/mallard/Sweden/105/2002_Sweden_H7N7
81
A/gadwall/Netherlands/1/2006_H9N2
90
A/Netherlands/219/2003_Netherlands_H7N7
88
100 A/chicken/Netherlands/2586/2003_Netherlands_H7N7
A/England/AV877/1996_United_Kingdom_H7N7
A/chicken/Israel/184/2009_H9N2
A/chicken/Korea/MS96-CE6/1996_H9N2
A/chicken/China/Guangxi14/2000_H9N2
A/chicken/Guangdong/6/97_H9N2
A/chicken/Pakistan/4/99_H9N2
100
A/chicken/Pakistan/UDL-03/2008_H9N2
A/duck/Zhejiang/12/2011_H7N3
A/swine/Hong_Kong/9/98_H9N2
88
100
A/swine/Guangxi/7/2007_H9N2
99
A/chicken/Yokohama/aq120/2001_H9N2
A/wild_bird/Korea/A9/2011_H7N9
100
A/chicken/Korea/A146/2009_H9N2
A/chicken/Guangxi/55/2005_H9N2
A/duck/Nanchang/4-361/2001_H9N2
A/chicken/Shandong/KD/2009_H9N2
100
A/chicken/Hebei/A/2007_H9N2
78
A/duck/Fujian/FQ2/2007_H9N2
83
A/swine/Guangxi/S11/2005_H9N2
A/chicken/Yokohama/aq134/2002_H9N2
A/turkey/Minnesota/1/1988_H7N9
A/Canada/rv504/2004_Canada_H7N3
A/New_York/107/2003_USA_H7N2
A/ruddy_turnstone/Delaware_Bay/220/1995_H7N9
A/northern_shoverl/Mississippi/11OS145/2011_H7N9
A/blue-winged_teal/Ohio/566/2006_H7N9
A/ruddy_turnstone/DE/1538/2000_H7N9
A/Mexico/InDRE7218/2012_Mexico_H7N3
A/blue-winged_teal/Guatemala/CIP049-01/2008_H7N9
A/blue-winged_teal/Guatemala/CIP049-02/2008_H7N9
100
98
PB2进化树
红色标记为本次病例
●H7 人感染病例
100
70
99
90
86
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
0.02
检测方法
• 2010年及之前下发的H7引物不再用于
检测。
• 今天下发新的H7和N9检测引物（realtime PCR）。
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
检测策略
• 不明原因肺炎病例标本检测：
建议各流感监测网络实验室收到不明原因肺炎标
本后，要在12小时内进行标本处理和核酸检测，
同时进行甲型通用引物，甲型H1N1流感病毒，季
节性H3N2流感，A（H5）、A (H7N9)亚型流感病
毒的检测。如果H5或H7亚型检测阳性，在生物安
全三级实验室进行流感病毒的分离鉴定，无病毒
分离条件的实验室，将标本在24小时内送国家流
感中心进行复核鉴定和病毒分离。
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION
谢谢！
CHINESE CENTER FOR DISEASE CONTROL AND PREVENTION