中药多糖增强猪蓝耳病卵黄抗体效价研究  

Chinese herbal medicine polysaccharide enhancing egg-yolk antibody titer of PRRS

作　　者：陈晓兰[1,2] 杨海峰[1] 贾纪萍[1] 陈兴颖[2] 蒋春茂[1] 王德云[2] 胡元亮[2] 

机构地区：[1]江苏农牧科技职业学院,江苏泰州225300 [2]南京农业大学动物医学院,江苏南京210095

出　　处：《中国兽医学报》2015年第9期1495-1500,共6页Chinese Journal of Veterinary Science

基　　金：江苏省自然科学基金面上资助项目(BK2011537);江苏省农业三新工程资助项目(SXGC[2013]291)

摘　　要：为探讨不同中药多糖对猪蓝耳病（PRRS）卵黄抗体效价的影响,本试验采用水提法制备卵黄水提液,2次盐析法纯化卵黄抗体（IgY）,SDS-PAGE法检测精制卵黄抗体的纯度。采用棋盘法确定了抗原最佳包被浓度和酶标二抗最佳工作浓度,建立了PRRS特异性卵黄抗体ELISA检测方法。并用PRRS疫苗4种不同免疫剂量免疫蛋鸡,确定最佳免疫方法。选择桑叶多糖、杜仲多糖、桑叶杜仲多糖、地黄多糖和藿蜂酮注射液饮水给药与PRRS疫苗联合免疫蛋鸡,通过卵黄抗体效价变化考察不同中药多糖的免疫增强作用。结果显示：选择抗原的最佳包被浓度为10-9,酶标二抗的最佳工作浓度为1：5 000,卵黄WFS的最佳稀释度为2-6;鸡肌注0.2mL/羽PRRS疫苗,在免疫后2~6周产生的抗体效价最高;鸡在免疫的同时以4mg/羽的剂量饮水给以桑叶杜仲复方多糖,连续3d,在免疫后的第2~9周卵黄抗体效价均最高,优于其他各组。以上结果表明,桑叶杜仲复方多糖饮水给药配合疫苗一起使用能显著提高PRRS卵黄抗体效价,可作为免疫增强剂使用。In order to discuss the effect of Chinese herbal medicine polysaccharide enhancing PRRS egg yolk antibody,egg yolk water soluble fraction（WFS） was prepared by water extraction, twice salting-out method was used to purify egg yolk antibody （IgY） ,and the purity of refined yolk anti- body IgY was detected by SDS-PAGE. The best concentration of coating antigen and the second enzyme-labeled antibody were comfirmed by chess board method,and ELISA method was estab- lished to determine PRRS specificity yolk antibody titer. Laying hens were vaccinated with four PRRS vaccine doses to determine the best immune method. When laying hens were vaccinated with PRRS vaccine, mulberry leaves polysaccharide （MP）, eucommia ulmoides polysaccharide （EUP）, mulberry leaves and eucommia ulmoides composite polysaccharide（MP-EUP）,rehmannia polysac- charide （RGPS） and epimedium polysaccharide-propolis flavone injection（EPI） were administered by drinking to compare the immune enhancement effects of Chinese herbal medicine polysaccha- rideby by the changes of specific IgY antibody titer. The results were as follows：the best concen- tration of the coated antigen and the second antibody were 1 ： 109 and 1 ： 5 000,and the best di- lution multiple of WFS was 2.6 ;antibody titer at 2 w to 6 w after vaccination was the highest when chickens were intramuscularly injected with 0.2 mL/feather PRRS vaccine; egg yolk anti- body titer was the highest at 2 w to 9 w after vaccination and better than other groups whenchickens were administered orally with mulberry leaves and eucommia compound polysaccharide at the dose of 4 mg/feather for three successive days. The above results showed that the mulberry leaves and eueommia compound polysaccharide administered orally with PRRS vaccine could sig- nificantly improve the egg-yolk antibody titer of PRRS, and could be produced as immunopotentia- tor.

关 键 词：猪蓝耳病 卵黄抗体 中药多糖 效价 ELISA 

分 类 号：S852.65[农业科学—基础兽医学]