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Development and evaluation of baculovirus-expressed Chikungunya virus E1 envelope proteins for serodiagnosis of Chikungunya infection

机译:杆状病毒表达的基孔肯雅病毒E1包膜蛋白的开发和评估用于基孔肯雅感染的血清诊断

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Population-based serosurveillance studies provide critical estimates on community-level immunity and the potential for future outbreaks. Currently, serological assays, such as IgG enzyme-linked immunosorbent assays (ELISAs) and indirect immunofluorescence tests (IIFT) based on the inactivated whole virus are used to determine past Chikungunya virus (CHIKV) infection. However, these commercially available tests have variable sensitivities. To develop and evaluate recombinant based CHIKV-specific IgG antibody capture ELISAs (GAC-ELISAs), baculoviruses carrying wild-type (E1-A226, named WT) or mutant (E1-A226V, named MUT) E1 envelope protein genes of CHIKV were generated. The seroreactivity of recombinant CHIKV WT and MUT envelope proteins were determined using residual blood, collected from CHIKV-confirmed patients. The sensitivities of both recombinant CHIKV envelope proteins were 83.0% as measured by GAC-ELISAs. The specificities of both recombinant proteins were 87.8%. These GAC-ELISAs were also able to detect the persistence of anti-CHIKV IgG antibodies up to 6 months after the disease onset, together with rise in sensitivities with increasing time. These results suggest that the baculovirus purified recombinant CHIKV envelope proteins react with anti-CHIKV IgG antibodies and may be useful in population-based seroprevalence surveys. In addition, these GAC-ELISAs offer good diagnostic value to determine the recent/past CHIKV infection status in non-endemic populations
机译:基于人群的血清监测研究提供了有关社区一级免疫力和未来爆发可能性的重要估计。当前,基于灭活的全病毒的血清学测定,例如IgG酶联免疫吸附测定(ELISA)和间接免疫荧光测定(IIFT),被用于确定过去的基孔肯雅病毒(CHIKV)感染。但是,这些市售的测试灵敏度各不相同。为了开发和评估基于重组的CHIKV特异性IgG抗体捕获ELISA(GAC-ELISAs),产生了带有野生型(E1-A226,称为WT)或突变体(E1-A226V,称为MUT)的杆状病毒CH1的E1包膜蛋白基因。重组CHIKV WT和MUT包膜蛋白的血清反应性是使用从CHIKV确诊的患者中收集的残留血液确定的。通过GAC-ELISA测定,两种重组CHIKV包膜蛋白的敏感性为83.0%。两种重组蛋白的特异性均为87.8%。这些GAC-ELISAs还能够在疾病发作后长达6个月的时间内检测出抗CHIKV IgG抗体的持久性,以及随着时间的延长敏感性提高。这些结果表明,杆状病毒纯化的重组CHIKV包膜蛋白与抗CHIKV IgG抗体反应,可能在基于人群的血清阳性率调查中有用。此外,这些GAC-ELISAs对确定非流行人群的近期/过去CHIKV感染状况具有良好的诊断价值。

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