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目的:原核表达柯萨奇病毒A组5型(coxsackievirus A5，CV-A5)VP1蛋白，并制备抗VP1多克隆抗体(多抗)，为CV-A5相关定性定量研究制备试剂。方法:逆转录PCR扩增N端截短的CV-A5 n VP1-ΔN56后，克隆至原核表达载体pGEX-6P-1，获得pGEX-6P-1-VP1-ΔN56。将其转化大肠埃希菌BL21(DE3)，表达重组蛋白并纯化。用纯化的谷胱甘肽巯基转移酶-VP1-ΔN56融合蛋白经背部皮下免疫日本大耳白兔，制备多抗。n 结果:重组表达载体构建成功，融合蛋白以不可溶包涵体存在。ELISA、蛋白质印迹法检测表明，获得的兔多抗效价为10n 7，可特异性识别重组和天然CV-A5 VP1蛋白。n 结论:成功制备重组CV-A5 VP1蛋白及特异性多抗。为中和抗原表征研究及VP1定性定量分析奠定了基础。“,”Objective:To prokaryotically express coxsackievirus A5 (CV-A5) VP1 protein and prepare anti-VP1 polyclonal antibody as reagents for qualitative and quantitative CV-A5 study.Methods:An N-terminally-truncated fragment, CV-A5 n VP1-ΔN56, was amplified by reverse transcription PCR and cloned into prokaryotic expression vector pGEX-6P-1. The plasmid pGEX-6P-1-VP1-ΔN56 was transformed inton E. n coli BL21(DE3), and the recombinant protein with an N-terminally-tagged glutathione S-transferase (GST) was induced. The Japanese white rabbit was immunized with purified GST-VP1-ΔN56 fusion protein through subcutaneous injection on the back.n Results:The recombinant expression vector was successfully constructed, and the fusion protein was mainly insoluble in inclusion body. The specificity of the rabbit anti-GST-VP1-ΔN56 polyclonal antibody was characterized using recombinant VP1 and natural particles by ELISA and Western blot, and the antibody titer was 10n 7.n Conclusions:The recombinant protein and rabbit anti-GST-VP1-ΔN56 polyclonal antibody specifically recognizing both recombinant and natural VP1 are produced successfully. The prepared antibody could be used for characterization of the CV-A5 neutralizing antigen, as well as for the qualitative and quantitative analysis of VP1.