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根据汉坦病毒Ｈ８２０５株ＮＰ基因的序列，设计一对引物，扩增ＮＰ前２９２个氨基酸多肽基因片段，克隆于表达载体ｐＧＥＸ３Ｘ，与载体中２６ｋＤ的谷胱苷肽巯基转移酶（ＧＳＴ）融合表达。ＳＤＳＰＡＧＥ显示表达产物（ＧＳＴＮＰ）主要以包涵体形式存在。Ｗｅｓｔｅｒｎｂｌｏｔｉｎｇ表明此融合蛋白有抗原性。包涵体经分离、洗涤、溶解后，Ｓｅｐｈａｒｏｓｅ６Ｂ层析纯化，用此融合蛋白作抗原，进行ＥＬＩＳＡ法检测临床ＨＦＲＳ病人标本的ＩｇＧ和ＩｇＭ，有很好的特异性和敏感性。有生物活性的汉坦病毒Ｈ８２０５ＮＰ的体外表达成功，为汉坦病毒基因工程抗原的大量制备奠定了基础，也为汉坦病毒的临床检测和流行病学调查提供了一种廉价、安全、可靠的抗原。
According to the sequence of NP gene of Hantavirus H8205 strain, a pair of primers was designed to amplify the 292 amino acid polypeptide fragments of NP before cloning in expression vector pGEX3X and glutathione S-transferase (GST) of 26 kD in vector. Fusion expression. SDS PAGE showed that the expression product (GST NP) mainly in the form of inclusion bodies. Western blotting showed that the fusion protein has antigenicity. Inclusion bodies were isolated, washed, lysed and purified by Sepharose 6B chromatography. The fusion protein was used as antigen to detect IgG and IgM in clinical samples of HFRS patients with good specificity and sensitivity. The successful expression of Hantaan virus H8205NP with biological activity laid the foundation for the preparation of genetically engineered antigen of Hantavirus and provided a cheap, safe and reliable method for the clinical detection and epidemiological investigation of Hantavirus. antigen.