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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: ; ; ; ; ; ; (谭志坚);

机构地区: 华南农业大学兽医学院

出  处: 《中国畜牧兽医》 2011年第8期52-55,共4页

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 抗体应答

领  域: [农业科学] [农业科学] [农业科学]

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