摘要
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。
Objective To use recombinant protein of lymphocytic choriomeningitis virus (LCMV) as alternative antigen to whole virus. Methods\ LCMV sRNA site 1816-2178 nucleotides coding Np site 380-500 amino acid (LN380-500) was synthesized. A recombinant plasmid pET LN380-500 was constructed and expressed in E.coli JM109 (DE3). The expressed LN380-500 protein was purified by Ni NTA Agarose and established high specificity in ELISA for the detection of antibody to LCMV. Results\ The purified 38×10 3 protein and whole virus were applied in ELISA for the detection of LCMV antibody in mice, golden hamsters and guinea pigs. The results showed no different in general, and recombinant protein ELISA without the preparation of control antigen was higher specificity and more simple. Conclusion\ The ELISA for detection of antibody to LCMV with synthetic gene expression as alternative antigen for lymphocytic choriomeningitis virus not only excluded possible spread of virus,but also enhanced the specificity.
出处
《中国实验动物学报》
CAS
CSCD
2001年第4期209-212,共4页
Acta Laboratorium Animalis Scientia Sinica
基金
卫生部科学研究基金 ( 98 2 0 49)
上海市科技发展基金 ( 9949190 31)
