| 间接ELISA检测鸭疫里默氏杆菌血清抗体方法的建立 |
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| 引用本文： | 李富祥,杨斌,常志顺,王传禹,姚俊,刘艳丽,李华春. 间接ELISA检测鸭疫里默氏杆菌血清抗体方法的建立[J]. 中国畜牧兽医, 2010, 37(1): 202-205 |
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| 作者姓名： | 李富祥 杨斌 常志顺 王传禹 姚俊 刘艳丽 李华春 |
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| 作者单位： | (1.云南省热带亚热带动物病毒病重点实验室， 昆明 650224； 2.云南省畜牧兽医科学院养禽与禽病研究所， 昆明 650224) |
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| 基金项目： | 科研院所技术开发研究专项(2007CF009) |
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| 摘 要： | 应用1型鸭疫里默氏杆菌(云南株)超声粉碎物作为包被抗原,建立检测鸭疫里默氏杆菌血清抗体的间接ELISA方法。用倍比稀释法确定HRP标记羊抗鸭二抗最佳稀释倍数为1∶3500,并用棋盘测定法确定抗原的最佳包被浓度为2.7mg/mL,血清最佳稀释倍数为1∶300,阴性血清临界值为0.381;阻断试验结果表明,1型鸭疫里默氏杆菌与其抗血清阻断阳性,与鸭大肠杆菌O78、O132菌株和FJ4型鸭疫里默氏杆菌阻断阴性(无交叉反应)。重复性试验结果表明,该ELISA方法批内变异系数≤0.246,批间变易系数≤0.889。结果表明,该ELISA方法特异性强,重复性好,可用于1型鸭疫里默氏杆菌(云南株)血清抗体的检测。
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| 关 键 词： | 鸭疫里默氏杆菌 ELISA 血清抗体 ELISA效价 |
Development and Application of Indirect ELISA for the Detection of Serum Antibodies against Riemerella nnntipestifer in Duck |
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| LI Fu-xiang,YANG Bin,CHANG Zhi-shun,WANG Chuan-yu,YAO Jun,LIU Yan-li,LI Hua-chun. Development and Application of Indirect ELISA for the Detection of Serum Antibodies against Riemerella nnntipestifer in Duck[J]. China Animal Husbandry & Veterinary Medicine, 2010, 37(1): 202-205 |
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| Authors: | LI Fu-xiang YANG Bin CHANG Zhi-shun WANG Chuan-yu YAO Jun LIU Yan-li LI Hua-chun |
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| Affiliation: | （1.Yunnan Tropical and Subtropical Animal Virus Disease Laboratory, Kunming 650224,China; 2.Institute for Poultry Science & Health of Yunnan Animal Science and Veterinary Institute, Kunming 650224, China） |
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| Abstract: | Indirect ELISA was developed and applied for detection of serum antibodies against serotype 1(Yunnan strain)Riemerella anatipestifer (RA) in ducks, using ultrasonic RA as the coating antigen. Dilution method shew the optimum HRP-Goat Anti-Duck IgG dilution was 1∶3500, chessboard test shew the optimum coating antigen concentration was 2.7 mg/mL,the optimum dilution of serum samples was 1∶300 and the cut-off value of negative serum was 0.381.The interruption test shew the reactions between antiserum of serotype 1 RA and the coating antigen could be interrupted by specific type 1 RA, but could not be interruped by E.coli serotype O78 and serotype O132 strains(no cross-reaction).Repetition test shew the variant coefficients of the D value within assay and between assays less than 0.246 and 0.889 respectively. An interruption test and repetition test showed that the ELISA assay had good specificity and reproducibility to detect serum antibodies against serotype 1 RA in ducks. |
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| Keywords: | Riemerella anatipestifer ELISA serum antibody ELISA titer |
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