鸭坦布苏病毒病灭活疫苗血清学试验与免疫攻毒保护试验的相关性

doi:10.3864/j.issn.0578-1752.2019.23.023

摘要/Abstract

摘要：

【目的】评价鸭坦布苏病毒病灭活疫苗2种效力评价方法平行关系,为采用血清学检验技术替代鸭坦布苏病毒病灭活疫苗本动物攻毒效力检验奠定基础。【方法】3批疫苗实验室制品分别以0.15、0.3、0.5、1.0 mL经肌肉注射途径免疫51日龄青年鸭和260日龄产蛋鸭,首次免疫后14 d,按相同的剂量和途径进行二次免疫。二免后28日采血,分离血清并测定血清的HI抗体效价,根据抗体效价对试验鸭分组,并分别经肌肉注射途径接种0.5 mL（100 DID₅₀）的鸭坦布苏病毒。攻毒后2 d采血,分离病毒,产蛋鸭攻毒后8 d剖检,观察卵巢病变。比较和分析HI抗体效价和攻毒保护结果的相关性。【结果】免疫青年鸭HI抗体效价≤1﹕5、1﹕10、1﹕20、1﹕40、1﹕80、1﹕160、1﹕320、1﹕640组保护率分别为50%（20/40）、43.8%（7/16）、74.3%（26/35）、86.7%（26/30）、96.6%（57/59）、92.7%（51/55）、95%（38/40）和100%（11/11）,HI抗体效价低于1﹕20时攻毒保护率为48.2%,效价1﹕20时攻毒保护率为74.3%,效价1﹕40及以上时攻毒保护率达93.8%。免疫产蛋鸭HI抗体效价≤1﹕5、1﹕20、1﹕40和1﹕80组保护率分别为66.7%（12/18）、83.3%（10/12）、95.2%（20/21）和100%（15/15）;HI抗体效价1﹕10、1﹕160、1﹕320和1﹕640组保护比例分别为2/3、7/7、1/1和2/2;HI抗体效价为1﹕10及以下时攻毒保护率为66.7%,效价1﹕20时攻毒保护率为83.3%,效价1﹕40及以上时攻毒保护率达97.8%。青年鸭和产蛋鸭的HI抗体效价与攻毒保护率之间均有极显著的正相关性,Spearman相关系数分别为0.905（P<0.01）和0.932（P<0.01）。【结论】鸭坦布苏病毒病灭活疫苗（HB株）免疫后HI抗体与攻毒保护率存在正相关,可替代免疫攻毒法用于鸭坦布苏病毒病灭活疫苗的效力评价,初步将HI抗体效价1﹕20定为鸭坦布苏病毒病灭活疫苗免疫后产生抗体保护的标准。

关键词: 鸭坦布苏病毒, HI试验, 抗体效价, 免疫攻毒, 相关性

Abstract:

【Objective】The objective of this paper was to evaluate the correlation of the serological test (HI) and challenge test of duck Tembusu virus inactivated vaccine.【Method】Three batches of vaccines ( laboratory trial products) were used to immunize the 51-day-old young ducks and 260-day-old laying ducks by intramuscular injection in 0.15, 0.3, 0.5 and 1.0 mL, respectively. Secondary immunization was proceeded in the same doses and route at 14 day after primary immunization. Ducks were regrouped according to the HI titer and challenged with 0.5 mL (100 DID₅₀) DTMUV at day 28 after secondary immunization. Serum samples were collected at 2 days later for virus isolation and laying ducks were dissected at 8 days later to observe the ovarian lesions. All the results were counted to evaluate the immune potency of vaccines. The correlation of HI titer and immune protection rate was analyzed.【Result】The protection rate of immunized young ducks with HI titer of 1:5 or lower, 1:10, 1:20, 1:40, 1:80, 1:160, 1:320, 1:640 was 50% (20/40), 43.8% (7/16), 74.3% (26/35), 86.7% (26/30), 96.6% (57/59), 92.7% (51/55), 95% (38/40) and 100% (11/11), respectively. The protection rate was 48.2% with HI titer lower than 1:20, while the rate was 74.3% with HI titer of 1:20 and 93.8% with HI titer of 1:40 or greater. In laying ducks, the protection rate of flocks with HI titer of 1:5 or lower, 1:20, 1:40 and 1:80 was 66.7% (12/18), 83.3% (10/12), 95.2% (20/21) and 100% (15/15), respectively. The protected proportion of ducks with HI titer of 1:10, 1:160, 1:320 and 1:640 was 2/3, 7/7, 1/1 and 2/2, respectively. The protection rate was 66.7% with HI titer of 1:10 or below, while the rate was 83.3% with HI titer of 1:20 and 97.8% with HI titer of 1:40 or greater. There were significant positive correlations between the HI titer and the immune protection rate both in young ducks and laying ducks. The Spearman’s correlation coefficient was 0.905 (P<0.01) and 0.932 (P<0.01), respectively. 【Conclusion】There were positive correlations between the HI titer and the immune protection rate of duck Tembusu virus inactivated vaccine (strain HB). The HI detection could be used to evaluate the efficacy of duck Tembusu virus inactivated vaccine (strain HB) instead of challenge test. The HI titer of 1:20 could be defined as the criteria of protection for the duck Tembusu virus inactivated vaccine after immunization.

Key words: duck Tembusu virus, HI test, antibody titer, challenge, correlation

王小蕾,林健,杨志远,刘立新,段会娟,程慧敏,赵际成,潘洁,刘月焕. 鸭坦布苏病毒病灭活疫苗血清学试验与免疫攻毒保护试验的相关性[J]. 中国农业科学, 2019, 52(23): 4423-4428.

WANG XiaoLei,LIN Jian,YANG ZhiYuan,LIU LiXin,DUAN HuiJuan,CHENG HuiMin,ZHAO JiCheng,PAN Jie,LIU YueHuan. Correlation Research of the Serological Test (HI) and Challenge Test of Duck Tembusu Virus Inactivated Vaccine[J]. Scientia Agricultura Sinica, 2019, 52(23): 4423-4428.

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疫苗 Vaccines	免疫鸭日龄 Day age of immunized ducks	免疫剂量（mL） Doses of immunization	免疫途径 Immunization route	免疫鸭数量 No. of immunized ducks
实验室制品01 Laboratory trial products 01	51	0.15 / 0.3 / 0.5 / 1	胸肌注射 Intramuscular injection	100 (25 / 25 / 25 / 25)
实验室制品01 Laboratory trial products 01	260	0.3 / 0.5 / 1		30 (10 / 10 / 10)
实验室制品02 Laboratory trial products 02	51	0.15 / 0.3 / 0.5 / 1		100 (25 / 25 / 25 / 25)
实验室制品02 Laboratory trial products 02	260	0.3 / 0.5 / 1		30 (10 / 10 / 10)
实验室制品03 Laboratory trial products 03	51	0.15 / 0.3 / 0.5 / 1		96 (24 / 24 / 24 / 24)
实验室制品03 Laboratory trial products 03	260	0.3 / 0.5 / 1		30 (10 / 10 / 10)
对照组	51	/	/	40
Control	260	/	/	20

分组 Groups	青年鸭保护率（%） Protection rate of young ducks	产蛋鸭保护率（%） Protection rate of laying ducks
≤1:5	50 (20^A / 40^B)	66.7 (12 / 18)
1:10	43.8 (7 / 16)	2 / 3
1:20	74.3 (26 / 35)	83.3 (10 / 12)
1:40	86.7 (26 / 30)	95.2 (20 / 21)
1:80	96.6 (57 / 59)	100 (15 / 15)
1:160	92.7 (51 / 55)	7 / 7
1:320	95 (38 / 40)	1 / 1
1:640	100 (11 / 11)	2 / 2
合计 Total	82.5 (236 / 286)	87.3 (69 / 79)
对照组 Control	0% (0 / 40)	10% (2 / 20)