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，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 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，制备多抗体。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10⁷，可特异性识别重组和天然CV-A5 VP1蛋白。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定性定量抗体分析奠定了基础。

Objective  To prokaryotically express coxsackievirus A5 (CV-A5) VP1 protein and prepare polyclonal antibody as quantitative reagents for CV-A5 study . Methods  An N-terminally-truncated fragment, CV-A5-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 into E. 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. 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 recombinant VP1 prokaryotic expressed and natural particles by ELISA and Western blot,and the antibody titer was 10⁷ . Conclusions The recombinant protein and rabbint 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 qualitative and quantitative analysis of VP1.