蛋鸡肠道有益菌的分离与初步鉴定

为了分离蛋鸡肠道中存在的有益菌,试验采用常规分离方法对产蛋高峰期蛋鸡肠道中有益菌进行分离鉴定,测定了目标菌株的理化性质、体外抑菌效果,并进行了雏鸡安全性试验。结果表明:从蛋鸡空肠中分离出乳酸菌4株,分别为K1、K2、K3、K4;从盲肠中分离出乳酸菌1株,为M1;从盲肠中分离芽孢杆菌3株,分别为Y1、Y2、Y3。通过系列生理生化鉴定,初步确定以上乳酸菌依次为植物乳杆菌、嗜酸乳杆菌、德氏乳杆菌德氏亚种、德氏乳杆菌乳亚种和短乳杆菌,芽孢杆菌依次为枯草芽孢杆菌、蜡样芽孢杆菌和地衣芽孢杆菌。乳酸菌对大肠杆菌O2有较强的抑菌作用,K1、K2、K3、K4、M1的抑菌圈直径分别为18.30,16.07,11.45,17.26,10.26 mm;而芽孢杆菌抑菌作用相对较弱,Y1、Y2、Y3的抑菌圈直径分别为9.25,8.46,8.37 mm。雏鸡日饮活菌20亿个为安全剂量。

Comparative Studies on Detection of Antibodies against Infectious Bursal Disease Virus with Test Strips and Agar Gel Immunodiffusion Method

[Objective] This study aimed to compare the detection results of antibodies against infectious bursal disease virus with test strips and agar gel immunodiffusion method. [Method] Antibodies against infectious bursal disease virus in chicken serum were detected using test strips developed in our laboratory, and the results were comparad^with that using traditional agar diffusion method. [Result] The comparative study of the two methods showed that the sensitivity of test strips was eight times over agar gel immunodiffusion; test strips showed higher detection rate in the deter- mination test of 216 clinical samples, with high specificity, easy preservation, and simple and rapid operation, thereby being more suitable for the monitoring of clinical antibodies. [Conclusion] Test strips could replace the existing serological methods, having great promotion and application value in antibody monitoring.

基于口蹄疫病毒非结构蛋白3AB/2C的ELISA鉴别检测试剂盒的研制与应用

基于口蹄疫病毒（FMDV）非结构蛋白3AB/2C,研制快速鉴别口蹄疫自然感染和疫苗接种的ELISA检测试剂盒。诱导表达3AB/2C蛋白,His亲和层析纯化目的蛋白;以纯化的3AB/2C蛋白为包被抗原,建立间接ELISA方法,优化ELISA反应条件;标准化ELISA操作规程和试剂盒组分,组装ELISA试剂盒,进行特异性、敏感性、符合率、重复性、血清交叉反应、保存期等性能评价;分离临床牛血清进行应用性检测,评价ELISA试剂盒适用性。结果获得了大小约51 ku可溶性表达3AB/2C蛋白,亲和层析纯化后纯度可达90%以上;确定最适抗原包被浓度为3.25μg/m L,血清稀释度1∶80,封闭液为2%乳清蛋白,封闭条件为37℃1 h 4℃10 h;试剂盒敏感性97.5%、特异性98.3%,批内、批间变异系数分别为3.40%～6.71%和5.20%～8.44%;与3种商品化试剂盒符合率分别为98.90%、98.02%和96.72%;试剂盒4℃条件下保存期可达12个月;1 457份临床样品检测,FMDV感染阳性率6.41%～30.35%。本研究研制的鉴别ELISA试剂盒检测结果可靠、稳定性好,能有效鉴别FMDV自然感染和疫苗免疫,可为FMD综合防控提供技术支撑。