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目的 用重组蛋白抗原取代淋巴细胞脉络丛脑膜炎病毒 (LCMV)抗原用于实验动物感染LCMV检测。方法 合成LCMVsRNA第 1816～ 2 178位核苷酸序列编码的Np第 380～ 5 0 0位氨基酸基因 ,克隆在Hisx6- GSTpET 2 8a载体中 ,表达产物经固定化金属配体亲和层析纯化 ,建立ELISA检测试剂。结果 合成基因表达产物经Ni NTA Agarose纯化后纯度达到 95 %以上。ELISA检测小鼠、地鼠和豚鼠结果与全病毒抗原基本一致 ,而且特异性较强 ,操作简便。结论 用合成基因表达产物取代LCMV抗原检测病毒抗体 ,不仅可以消除病毒传播可能 ,而且特异性强。为实验动物及生物材料质量控制提供安全有效方法
Objective To replace the LCMV antigen with recombinant protein antigen for LCMV infection in laboratory animals. Methods The amino acid sequence of 380th to 500th of Np encoded by nucleotides 1816-278 of LCMVsRNA was synthesized and cloned into Hisx6-GSTpET 2 8a vector. The expressed product was purified by immobilized metal ligand affinity chromatography. Establish ELISA test reagent. Results The purity of the expressed product was over 95% after purification by Ni NTA Agarose. ELISA test mice, hamsters and guinea pig results with the whole virus antigen is basically the same, but the specificity is strong, easy to operate. Conclusion The detection of virus antibodies by replacing the LCMV antigen with synthetic gene expression products not only can eliminate the possibility of virus transmission, but also has strong specificity. Provide a safe and effective method for the quality control of laboratory animals and biological materials