检测鸡球虫四价弱毒疫苗抗体应答的ELISA方法的建立及初步应用

›› 2011, Vol. 38 ›› Issue (8): 52-55.

检测鸡球虫四价弱毒疫苗抗体应答的ELISA方法的建立及初步应用

李仁良¹, 袁子国¹, 侯杰¹, 谭志坚², 林瑞庆¹, 朱兴全³, 翁亚彪¹

1. 华南农业大学兽医学院,广东广州 510642;2. 佛山正典生物技术有限公司,广东佛山 528138;3. 中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,甘肃兰州 730046

收稿日期:2010-11-20 修回日期:1900-01-01 出版日期:2011-08-20 发布日期:2011-08-20
通讯作者: 翁亚彪

Establishment and Preliminary Application of an ELISA for the Detection of Antibody Responses in Chickens Induced by the 4-valent Attenuated Vaccine against Chicken Coccidiosis

LI Ren-liang¹, YUAN Zi-guo¹, HOU Jie¹, TAN Zhi-jian², LIN Rui-qing¹, ZHU Xing-quan³, WENG Ya-biao¹

1. College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China;2. Standard Bio-Tech Co., Ltd, Foshan 528138, China;3. State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China

Received:2010-11-20 Revised:1900-01-01 Online:2011-08-20 Published:2011-08-20

摘要/Abstract

摘要： 本研究旨在建立检测鸡球虫四价弱毒疫苗(柔嫩艾美耳球虫、毒害艾美耳球虫、堆型艾美耳球虫、巨型艾美耳球虫)抗体应答的酶联免疫吸附试验(ELISA),并初步阐明鸡免疫四价弱毒疫苗及攻虫后的抗体应答。将300羽岭南黄鸡分为A(免疫攻虫组)、B(不免疫不攻虫组)、C(不免疫攻虫组)共3个组。A组雏鸡在4日龄时进行首免,11日龄时鸡开始二免,二免完成后(17日龄时)对A组及C组的鸡进行攻虫(28万个4种球虫的混合卵囊/羽),7 d后检查A、C两组鸡的存活率。在鸡二免和三免完成后,分别对A、B组进行采血,通过优化最佳抗原包被浓度、酶结合物工作浓度及最佳血清稀释度,建立了ELISA方法来检测鸡体对球虫四价弱毒疫苗的抗体应答情况。A组鸡血清抗体的平均D值为0.870和0.904,明显高于B组鸡的平均D值0.261和0.270,证明了鸡体免疫疫苗后可以刺激产生相应抗体。免疫鸡攻虫后的存活率和抗体应答的检测结果,都充分说明了此鸡球虫弱毒四价疫苗具有良好的免疫原性,所建立的ELISA方法为今后评价鸡球虫弱毒四价疫苗的免疫效力提供了有效手段。

关键词: 鸡球虫弱毒四价疫苗; 鸡; ELISA; 抗体应答

Abstract: This study aimed to establish an enzyme-linked immunosorbent assay (ELISA) to examine antibody responses in chickens induced by a 4-valent attenuated vaccine (COCVAC) containing E. tenella, E. necatrix, E. acervulina and E. maxima against chicken coccidiosis. Three hundred Lingnan Yellow chickens were randomly divided into groups A (immunization-challenge group), B (non-immunization-non-challenge group) and C (non-immunization -challenge group), respectively; and each group had 100 broilers. Chickens in group A were immunized with the vaccine at the age of 4 days, and then boosted (second immunization) at the age of 11 days. After the second immunization chickens in groups A and C were challenged with 280000 mixed oocysts of the 4 Eimeria species/bird) at the age of 17 days. Seven days after challenge infection, the survival rate of in groups A and C was compared. After the second and the third immunizations, blood samples were collected from birds in groups A and B, and serum antibodies to the vaccine were detected using the established ELISA. The average optical density (D) values of birds in group A were 0.870 and 0.904 respectively, which were significantly higher than the average D values of chickens in group B (0.270 and 0.261 respectively). These results indicated that chickens immunized with the 4-valent attenuated coccidian vaccine can induce significant antibody responses, which correlates with the immune protection it induced, demonstrating that the established ELISA provides a useful tool for the evaluation of vaccine efficacy.

Key words: The 4-valent attenuated coccidian vaccine COCVAC; chicken; ELISA; antibody response

中图分类号:

S853.32

李仁良;袁子国;侯杰;谭志坚;林瑞庆;朱兴全;翁亚彪. 检测鸡球虫四价弱毒疫苗抗体应答的ELISA方法的建立及初步应用[J]. , 2011, 38(8): 52-55.

LI Ren-liang;YUAN Zi-guo;HOU Jie;TAN Zhi-jian;LIN Rui-qing;ZHU Xing-quan;WENG Ya-biao. Establishment and Preliminary Application of an ELISA for the Detection of Antibody Responses in Chickens Induced by the 4-valent Attenuated Vaccine against Chicken Coccidiosis[J]. China Animal Husbandry & Veterinary Medicine, 2011, 38(8): 52-55.

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