摘要
以痢疾志贺氏菌为抗原免疫产蛋母鸡,从鸡卵黄中提取免疫球蛋白,建立抗痢疾志贺氏菌的特异性IgY的效价检测方法,并研究母鸡的免疫应答性以及抗体的提取方法和体外抑菌效果。研究结果表明:用108cfu/mL和109cfu/mL剂量的抗原分别免疫的鸡的卵黄中,于初次免疫后的第12天出现抗痢疾志贺氏菌IgY,两组效价均为1∶3600。经加强免疫后效价迅速上升,至第48天低剂量免疫组达到最高效价1∶28800;第60天高剂量免疫组达到最高效价1∶57600。低剂量免疫组的效价维持了210d,高剂量免疫组的效价在免疫后360d时仍维持在1∶7200水平。用水稀释法、硫酸铵溶液分级盐析、SephadexG-25凝胶过滤以提取IgY,提纯后IgY的效价翻倍。SDS-PAGE鉴定抗体的纯度,电泳图谱中出现抗体的轻链和重链两条带。体外抑菌实验表明,IgY能抑制痢疾志贺氏菌的生长。
In this study, laying hens were divided into two groups and were immunized with two different antigen dose of Shigella dysenteriae, then the specific IgY was isolated from egg yolk. The method of measuring the antibody titer, the variation of antibody titer, the technique for isolation the antibody, and the inhibitory effects of IgY on the bacteria in vitro were studied. The results showed that the highest titer of IgY against Shigella dysenteria in the high and low dose group could reach to 1：57 600 and 1：28 800 respectively. The titer of IgY against Shigella dysenteria produced by the group of hens immunized with low dose of antigen could maintain 210 days, while the titer of IgY produced by the group of hens immunized with high dose of antigen could maintain more than 1 year. IgY was extracted from yolk by the water dilution method, sodium sulfate precipitation, and gel filtration. After purification, the titer of IgY against Shigella dysenteria was one time higher than that before purification. In the SDS-PAGE electrophoragram, two major bands were observed, which represented the heavy chain and the light chain. The results also demonstrated that the specific IgY could inhibit the growth of Shigella dysenteria, when the concentration of IgY reached to 10 mg/mL, the growth of bacteria was inhibited effectively.
出处
《中国食品学报》
EI
CAS
CSCD
2006年第1期299-302,共4页
Journal of Chinese Institute Of Food Science and Technology
