CAJ | 학술논문

　　目的分段表达并纯化肾综合征出血热病毒核蛋白，应用array-ELISA技术评价重组表达的核蛋白片段的诊断价值。方法构建肾综合征出血热病毒核蛋白表达质粒pET-32a（＋）/Pn、pET-32a（＋）/Pc，在大肠杆菌中诱导表达并用亲和层析法纯化，运用array-ELISA方法鉴定重组蛋白的检测特异性，检测结果与商品化胶体金试剂盒进行比较。结果在大肠杆菌中正确表达了肾综合征出血热病毒的核蛋白，亲和纯化得到较高纯度的蛋白质；应用array-ELISA技术从16份临床疑似血清样本中检出13份阳性血清，与商品化胶体金试剂盒一致性达到94％。结论获得的重组蛋白His-Pn可作为肾综合征出血热特异性诊断抗原之一；array-ELISA方法可以作为检测病毒抗体的有效手段。
　　목적분단표체병순화신종합정출혈열병독핵단백，응용array-ELISA기술평개중조표체적핵단백편단적진단개치。방법구건신종합정출혈열병독핵단백표체질립pET-32a（＋）/Pn、pET-32a（＋）/Pc，재대장간균중유도표체병용친화층석법순화，운용array-ELISA방법감정중조단백적검측특이성，검측결과여상품화효체금시제합진행비교。결과재대장간균중정학표체료신종합정출혈열병독적핵단백，친화순화득도교고순도적단백질；응용array-ELISA기술종16빈림상의사혈청양본중검출13빈양성혈청，여상품화효체금시제합일치성체도94％。결론획득적중조단백His-Pn가작위신종합정출혈열특이성진단항원지일；array-ELISA방법가이작위검측병독항체적유효수단。
Objective To express and purify the recombinant nucleoprotein fragments of hemor-rhagic fever with renal syndrome ( HFRS) virus and to evaluate their diagnostic efficacy by using array-ELISA technology .Methods The target genes encoding nucleoprotein fragments of HFRS virus were amplified by PCR, and then inserted into prokaryotic expression vectors to construct the recombinant plasmids of pET -32a (+)/Pn and pET-32a(+)/Pc.The plasmids were transformed into E.coli BL21 ( DE3) to induce the ex-pression of nucleoprotein fragments by IPTG and the expressed products were purified by affinity chromatog -raphy using Ni-NTA agarose.The specificity and sensitivity of the recombinant antigens were evaluated by the assay of array-ELISA using commercial colloidal gold assay kit as a comparison .Results The recombi-nant nucleoprotein fragments of HFRS virus were correctly expressed in E.coli and highly purified by affinity chromatography .Array-ELISA showed that 13 of 16 suspected serum samples were positive by using the His-Pn protein as diagnostic antigen , consistency with the commercial colloidal gold assay kit reaching 94%. Conclusion The recombinant His-Pn protein expressed in E.coli cells could be used for specific serodiag-nosis of HFRS virus as its high antigenicity and sensitivity .The array-ELISA is an effective assay for the de-tection of virus at protein level .