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目的制备高特异性长双歧杆菌多克隆抗体,为长双歧杆菌的免疫学检测提供参考。方法以破碎的长双歧杆菌细胞碎片为抗原免疫小鼠,制备抗血清;利用偶联相同抗原的磁珠,分离纯化长双歧杆菌多抗,分别采用间接ELISA、SDS-PAGE和点杂交法检测纯化多抗的效价、纯度和交叉反应性。结果所制备的纯化多抗效价约为1∶1500,回收率约为80%,纯度可达83%。纯化的多抗有效消除了与金黄葡萄球菌的非特异性交叉反应。结论已制备出高特异性的长双歧杆菌多克隆抗体,可用于长双歧杆菌制剂的菌体检测。
Objective To prepare highly specific polyclonal antibodies against Bifidobacterium longum and provide a reference for the immunological detection of Bifidobacterium longum. Methods Antibodies were prepared from the fragments of Bifidobacterium longum as antigen and antisera were prepared. The McAb against Bifidobacterium longum was isolated and purified by magnetic beads conjugated with the same antigen. The titer, purity and cross-reactivity of the purified polyclonal antibodies were tested. Results The titer of purified polyclonal antibody was about 1:1500, the recovery was about 80% and the purity was 83%. The purified polyclonal antibody effectively eliminates unspecific cross-reactivity with S. aureus. Conclusion Polyclonal antibodies against Bifidobacterium longum have been prepared and can be used for the detection of Bifidobacterium longum.