Huang J L, Huang J H, Shyu R H, Teng C W, Lin Y L, Kuo M D, Yao C W, Shaio M F
Division of Virology, Institute of Preventive Medicine, National Defense Medical Center, P.O. Box 90048-700, SanHsia, Taipei, Taiwan, Republic of China.
J Med Virol. 2001 Nov;65(3):553-60.
The prevalence of NS1 Ab response in patients with dengue viral infection and the potential of using recombinant NS1 protein as a diagnostic antigen for dengue viral infection were investigated. In this study, the full-length and C-terminal half of NS1 proteins (rNS1, rNS1-C) were highly expressed (10-30 mg/l) and further purified and refolded. The good antigenicity of the full-length rNS1 protein was confirmed by interaction with 19 dengue NS1-specific monoclonal antibodies (MAbs) in ELISA; however, the antigenicity of rNS1-C was relatively lower. The full-length rNS1 antigen also differentiated reliably between sera from dengue virus-infected patients and sera from normal controls. When rNS1 was used as an antigen to detect human anti-NS1 IgM and IgG Ab, the anti-NS1 Ab response was found in 15 of 17 patients (88%) with primary dengue infection and all 16 patients (100%) with secondary dengue infection. These results indicated that using the full-length rNS1 whose antigenicity is restored as ELISA antigen, a high anti-NS1 antibody prevalence could be detected in patients with either primary or secondary dengue infection. This finding suggested that the anti-NS1 antibody appeared not only in secondary and severe dengue virus infection and might not correlate the pathogenesis of dengue hemorrhagic fever. The study also verified that our purified rNS1 protein showed similar immunological properties as native dengue viral proteins. Genetic engineering production of recombinant NS1 antigen could provide a safe and valuable resource for dengue virus serodiagnosis.
研究了登革病毒感染患者中NS1抗体反应的流行情况以及使用重组NS1蛋白作为登革病毒感染诊断抗原的潜力。在本研究中，NS1蛋白的全长和C端半段（rNS1、rNS1-C）得到了高效表达（10 - 30mg/l），并进一步进行了纯化和复性。通过ELISA中与19种登革NS1特异性单克隆抗体（MAbs）的相互作用，证实了全长rNS1蛋白具有良好的抗原性；然而，rNS1-C的抗原性相对较低。全长rNS1抗原也能可靠地区分登革病毒感染患者的血清和正常对照的血清。当rNS1用作抗原检测人抗NS1 IgM和IgG抗体时，在17例原发性登革感染患者中有15例（88%）以及16例继发性登革感染患者全部（100%）检测到了抗NS1抗体反应。这些结果表明，将抗原性恢复的全长rNS1用作ELISA抗原，可以在原发性或继发性登革感染患者中检测到较高的抗NS1抗体流行率。这一发现表明，抗NS1抗体不仅出现在继发性和严重登革病毒感染中，可能与登革出血热的发病机制无关。该研究还证实，我们纯化的rNS1蛋白显示出与天然登革病毒蛋白相似的免疫特性。重组NS1抗原的基因工程生产可为登革病毒血清学诊断提供一种安全且有价值的资源。
