牛布鲁氏杆菌斑点酶联快速诊断新技术的研究

Study on New Technology for Rapid Diagnosis of Brucella abortus by DOT-ELISA

目的建立一种快速诊断牛布鲁氏杆菌病抗原的新方法。方法应用斑点酶联免疫吸附试验原理,将牛布鲁氏杆菌单克隆抗体与待检样品集成于一张纤维素膜上,与酶标二抗反应,再与底物液作用,通过肉眼观察底物液的颜色变化来判断待检样品中是否含有布鲁氏杆菌。结果该方法最低检测样品中活菌数为1万个菌落形成单位,整个检测过程需4.5个小时。结论牛布鲁氏杆菌斑点酶联快速诊断技术是一种简单、易行、特异(与标准大肠杆菌、沙门氏杆菌菌株没有交叉反应)、敏感(检测1万个单个菌)、省时(整个过程只需4.5个小时)的检测牛布鲁氏杆菌菌体的新方法,便于各级检验部门和基层广大农村牧区推广和应用。

Objective To establish a new rapid method for the diagnosis of Brucella abortus. Methods According to DOT - ELISA principle, the method is to integrate the McAb and the samples on a NC membrane. Reacting to HRP - antibody and the substrate, color changes of the substrate will determine whether the samples contain the brucella or not. Results The method could detect the brucella when samples containing only 10 000 CFU of brucella. The entire testing process would take 4.5 hours. Conclusions This DOT - ELISA is a simple, easy, differential （ without consensual reaction to the standard E. coli and Salmonella ）, sensitive （being sensitive to ten thousand bacilli ） and time -saving diagnosing technology comparing with ELISA. The whole process only takes 4.5 hours. It is convenient to apply in test departments of different levels and in the wide pastoral villages.