Establishment of an Indirect ELISA for Detecting Egg Yolk Antibody Against Chicken Flavivirus
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摘要: 为在检测产蛋鸡黄病毒抗体水平过程中减少应激,以鸡源黄病毒FQ-C1株为包被抗原,建立检测鸡源黄病毒卵黄抗体的间接ELISA方法。确定的优化条件是包被抗原浓度为5.7ng.μL-1,卵黄抗体以1∶160倍稀释,羊抗鸡IgG酶标抗体以1∶3 200稀释。经特异性、敏感性检验表明该方法特异性强、敏感性高,适合于产蛋鸡免疫后的抗体检测。Abstract: A novel Flavivirus infection,which mainly caused egg-dropping in ducks and chickens,has spread throughout China since 2010.To carry out the serological surveillance in chickens with minimum stress,an indirect enzyme-linked immuosorbent assay(ELISA) for detecting egg yolk antibody against Flavivirus in layer chickens has been developed by coating plate with chicken-origin Flavivirus FQ-C1 isolate.The optimal working concentration of antigen was 5.7 ng ·μL-1,the working concentration of egg yolk antibody was 1∶160 dilution and that of HRP-labeled goat anti-chicken IgG was 1∶3 200.After a color reaction of 10 minutes with Tetramethylbenzidine(TMB) liquid substrate,a threshold of positive/negative(P/N) ≥ 2.1 was determined as positive standard for the ELISA.The results indicated that the ELISA established by this research provided a high sensitivity and specificity test,which was suitable for the detection of antibodies against Flavivirus in immune laying hens.
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Key words:
- chicken-origin Flavivirus /
- indirect ELISA /
- egg yolk antibody
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[1] 曹贞贞,张存,黄瑜,等.鸭出血性卵巢炎的初步研究[J].中国兽医杂志,2010,46(12):3-6. [2] 万春和,施少华,程龙飞,等.一种引起种(蛋)鸭产蛋骤降新病毒的分离与初步鉴定[J].福建农业学报,2010,25(6):663-666. [3] SU J,LI S,HU X,et al.Duck Egg-Drop Syndrome Caused byBYD Virus,a New Tembusu-Related Flavivirus[J].PloS One,2011,6(3):e18106. [4] CAO Z,ZHANG C,LIU Y,et al.Tembusu virus in ducks,China[J].Emerg Infect Dis.,2011,17(10):1873-1875. [5] YAN P,ZHAO Y,ZHANG X,et al.An infectious disease ofducks caused by a newly emerged Tembusu virus strain inmainland China[J].Virology,2011,417(1):1-8. [6] 李玉峰,马秀丽,于可响,等.一种从鸭新分离的黄病毒研究初报[J].畜牧兽医学报,2011,42(6):885-891. [7] 胡旭东,路浩,刘培培,等.我国发现的一种引起鸭产蛋下降综合征的新型黄病毒[J].中国兽医杂志,2011,47(7):43-47. [8] 陈仕龙,陈少莺,王劭,等.一种引起蛋鸡产蛋下降的新型黄病毒的分离与初步鉴定[J].福建农业学报,2011,26(2):170-174. [9] 傅光华,黄瑜,施少华,等.鸡黄病毒的分离与初步鉴定[J].福建畜牧兽医,2011,33(3):1-2. [10] 黄欣梅,李银,赵冬敏,等.新型鹅黄病毒JS804毒株的分离与鉴定[J].江苏农业学报,2011,27(2):354-360. [11] 袁生,李金平,王敏儒,等.鹅源黄病毒的分离和初步鉴定[J].中国畜牧兽医,2012,39(9):47-50. [12] 施少华,傅光华,万春和,等.鹅源坦布苏病毒的分离及初步鉴定[J].中国兽医杂志,2011,48(12):10-11. -
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