鸭坦布苏病毒的血凝性

doi:10.3864/j.issn.0578-1752.2019.23.022

Abstract

Abstract:

【Objective】 The objective of this paper was to study the hemagglutinating activity of duck Tembusu virus (DTMUV). 【Method】The DTMUV was inoculated intracerebrally in the 1-3-day-old sucking mice of BALB/c in different dilution to observe the morbidity. The brains of sucking mice were collected and purified according to the referenced methods. The purified virus fluid was inactivated by suitable inactivator, followed by the inactivated inspection in 6-day-old SPF chicken embryo. The virus fluid was evaluated the hemagglutinating activity with the erythrocyte suspension of chicken, duck, goose, pigeon and swine. The concentration of the erythrocyte suspension used in the hemagglutination test was compared to select the proper concentration. The pH value of the reaction system between 6.0-7.0 and the reaction temperature including 4℃, room temperature and 37℃ were compared. 【Result】 In the 6 day after inoculation , the sucking mice inoculated with 1:10 dilution appeared severe clinical symptoms including twitch, paralysis, and most of them were in agonal stage, while the symptoms of the sucking mice inoculated with 1:50 and 1:100 dilution were slighter. The purified DTMUV fluid was pinkey and clarified. It could be inactivated by formaldehyde but not β-propiolactone that would produce lots of flocks and change the property of the fluid. The DTMUV could hemagglutinate with the erythrocyte suspension of chicken, duck, goose, pigeon and swine, and the agglutination graphics was clear and stable when the concentration of the erythrocyte suspension was 0.33%. The hemagglutinating activity of DTMUV could be showed when the reaction system pH value was 6.0-6.8, while the highest hemagglutination titer appeared during the pH 6.0-6.2. Furthermore, the hemagglutinating activity could be appeared at 4℃, room temperature and 37℃, respectively, although the time required for hemagglutination at 4℃ was much longer. 【Conclusion】The DTMUV proliferated in the sucking mice of BALB/c had a broad spectrum of hemagglutinating activity that could hemagglutinate with the erythrocyte suspension of chicken, duck, goose, pigeon and swine. The hemagglutinating activity was stable and could hemagglutinate with the goose erythrocyte at 4℃, room temperature and 37℃. The suitable reaction conditions were 0.33% concentration of the erythrocyte suspension and pH 6.0-6.2 of the reaction system.

Key words: duck Tembusu virus, hemagglutinating activity, reaction conditions

WANG XiaoLei, LIU YueHuan, DUAN HuiJuan, LIU LiXin, YANG ZhiYuan, ZHAO JiCheng, PAN Jie, LIU RuiHua, ZHAO WenQi, TIAN FangJie, Lü JinBao, LIN Jian. Hemagglutinating Activity of Duck Tembusu Virus[J].Scientia Agricultura Sinica, 2019, 52(23): 4415-4422.

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References 32

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稀释度 Dilution	接种数（只） No. of sucking mice	接种后不同时间（日）出现临床症状的鼠数 No. of mice appearing clinical symptoms at each day post inoculation
稀释度 Dilution	接种数（只） No. of sucking mice	1	2	3	4	5	6
1:10	20	N	N	N	N	A	C
1:50	193	N	N	N	N	B	C
1:100	120	N	N	N	N	B	C

灭活剂 Inactivator	终浓度 Final concentration (%)	灭活时间Inactivation hours (h)
灭活剂 Inactivator	终浓度 Final concentration (%)	24	48	72	96	120
甲醛 Formaldehyde	0.05	0^A/5^B	0/5	0/5	0/5	0/5
	0.1	0/5	0/5	0/5	0/5	0/5
	0.15	0/5	0/5	0/5	0/5	0/5
β-丙内酯 β-Propiolactone	0.0125	ND	ND	ND	ND	ND
	0.025	ND	ND	ND	ND	ND
	0.05	ND	ND	ND	ND	ND

动物 Animal	日龄 Age (d)	性别 Gender	HA效价 HA titer	凝集图形 Agglutination graphics
猪（长白猪） Swine (Landrace)	60	公母混合（3头） Mixed of male and female	1:256	红细胞对照不完全下沉,形成环状 The erythrocyte control sank not completely and formed a ring
鸡（SPF） Chicken (SPF chicken)	5	公（3只）Male	1:256	均一薄膜状凝集,清晰稳定 Clear and stable homogeneous thin-film agglutination
	360	公（3只）Male	1:256		均一薄膜状凝集,清晰稳定 Clear and stable homogeneous thin-film agglutination
	鸭（北京鸭） Duck (Peking duck)	60	母（3只）Female	1:512		均一薄膜状凝集,清晰稳定 Clear and stable homogeneous thin-film agglutination
	鸽（信鸽） Pigeon (Carrier pigeon)	360	公母混合（3只） Mixed of male and female	1:512	均一薄膜状凝集,清晰稳定 Clear and stable homogeneous thin-film agglutination
鹅（白鹅） Goose (White goose)	180	公母混合（3只） Mixed of male and female	1:512	均一薄膜状凝集,清晰稳定 Clear and stable homogeneous thin-film agglutination

鹅红细胞浓度 Concentration of goose erythrocyte	HA效价 HA titer	凝集图形 Agglutination graphics
0.2%	1:1024	红细胞对照孔下沉点小,结果不易判定 The erythrocyte control sank as a tiny point so that the results were difficult to interpretation
0.33%	1:512	均一薄膜状凝集,清晰稳定 Clear and stable homogeneous thin-film agglutination
0.5%	1:256	均一薄膜状凝集,清晰,有解凝迹象 Clear homogeneous thin-film agglutination
1%	1:128	均一薄膜状凝集,解凝快,不稳定 Clear homogeneous thin-film agglutination with fast dissolving

pH值 pH value	HA效价 HA titer	凝集图形 Agglutination graphics
6.0	1:512	均一薄膜状凝集,清晰稳定 Clear and stable homogeneous thin-film agglutination
6.2	1:512	均一薄膜状凝集,清晰稳定 Clear and stable homogeneous thin-film agglutination
6.4	1:256	均一薄膜状凝集,清晰稳定 Clear and stable homogeneous thin-film agglutination
6.6	1:256	均一薄膜状凝集,清晰稳定 Clear and stable homogeneous thin-film agglutination
6.8	1:128	均一薄膜状凝集,清晰稳定 Clear and stable homogeneous thin-film agglutination
7.0	1:64	易解凝,结果不易判定 The thin-film agglutination was easy to dissolve so that the result was difficult to interpretation

Hemagglutinating Activity of Duck Tembusu Virus

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反应温度 Reaction temperature	HA效价 HA titer	凝集图形 Agglutination graphics
37℃	1:512	均一薄膜状凝集,清晰稳定 Clear and stable homogeneous thin-film agglutination
室温Room temperature 4℃（60 min）	1: 512 /	均一薄膜状凝集,清晰稳定 Clear and stable homogeneous thin-film agglutination 红细胞对照沉降不完全,结果无法判定 The erythrocyte control sedimentated slowly so that the result was difficult to interpretation
4℃（过夜作用）（overnight）	1: 512	均一薄膜状凝集,清晰稳定 Clear and stable homogeneous thin-film agglutination

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