中文摘要：

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。

英文摘要：

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.