Position: Home > Articles > Establishment and Application of Sandwich ELISA Method for Detecting Avian Beta-defensin 6
Journal of Agricultural Science and Technology
2014,16
(4)
176-180
检测禽β-防御素6夹心ELISA法的建立及应用
作 者：
梁瑞;梁盈;祁克宗;王爱荣;彭开松
单 位：
安徽农业大学动物科技学院;河北省唐山市丰润区农业畜牧水产局
关键词：
AvBD6;夹心ELISA;ACTH;大肠杆菌;沙门氏菌
摘 要：
为定量检测禽β-防御素6(avian beta-defensin6,AvBD6),建立了AvBD6的双抗体夹心ELISA法,并应用于鸡血清中AvBD6浓度的检测。研究结果显示,包被抗体(鼠抗AvBD6 B细胞线性表位的多克隆抗体)、检测抗体(兔抗AvBD6氮端的多克隆抗体)和辣根过氧化酶(HRP)标记羊抗兔IgG的最佳工作稀释倍数分别为2 500、2 000和10 000,AvBD6在20~320 ng/mL的线性范围内,标准曲线回归方程为:y=12 024x-2 583,(R2=0.984 2)。对不同状态下鸡血清AvBD6浓度检测结果显示:ACTH(25 IU/kgBW)注射1 d后,对照组AvBD6浓度高于ACTH组;连续注射ACTH 5 d,对照组AvBD6浓度低于ACTH组,但二者间差异均不显著(P>0.05)。大肠杆菌感染后24 h和沙门氏菌感染后3 h、24 h,感染组AvBD6浓度均显著地高于对照组(P<0.05),表明AvBD6参与对大肠杆菌和沙门氏菌系统感染的抵抗。
译 名：
Establishment and Application of Sandwich ELISA Method for Detecting Avian Beta-defensin 6
作 者：
LIANG Rui;LIANG Ying;QI Ke-zong;WANG Ai-ronng;PENG Kai-song;College of Animal Science and Technology,Anhui Agricultural University;Fengrun District Agricultural Animal Husbandry and Fishery Bureau of Tangshan City;
关键词：
AvBD6;;sandwich ELISA;;ACTH;;Escherichia coli;;Salmonella
摘 要：
To detect avian beta-defensin 6(AvBD6) quantitatively,a sandwich ELISA method was established and applied to detect the concentration of AvBD6 in serum. The results showed that the optimal dilution of capture antibody( mouse anti-B cell linear epitopes of AvBD6 polyclonal antibody),detecting antibody( rabbit anti-Nterminal of AvBD6 polyclonal antibody) and horseradish peroxidase( HRP)-conjugated goat anti-rabbit IgG was2 500,2 000 and 10 000,respectively. The standard curve regression equation was y = 12 024x- 2 583,(R2= 0. 9842) in the linear detection range from 20 to 320 ng /mL. The serum AvBD6 concentration of control group was slightly higher than that of ACTH group( P > 0. 05) after 1 day of ACTH( 25 IU /kg BW) injection,but the AvBD6 concentration of the control group was slightly lower than that of ACTH group(P > 0. 05) after 5 d of ACTH injection;and the effect was not significant. The serum AvBD6 concentration of infected group after 24 h of Escherichia coli exposure or 3 h and 24 h of salmonella exposure were significantly higher than that of the control group(P < 0. 05).Our results indicated that AvBD6 involved in the resistance to system infection of Escherichia coli and salmonella.
