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  

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

作　　者：李仁良[1] 袁子国[1] 侯杰[1] 谭志坚 林瑞庆[1] 朱兴全[3] 翁亚彪[1] 

机构地区：[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

出　　处：《中国畜牧兽医》2011年第8期52-55,共4页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金项目(30901067);广东省科技计划项目(2009B020307011);广东省自然科学基金(10451064201005443)

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),7d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。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.

关 键 词：鸡球虫弱毒四价疫苗 鸡 ELISA 抗体应答 

分 类 号：S853.32[农业科学—临床兽医学]