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登革病毒包膜蛋白(E)肽的设计和异源表达及其在血清学诊断中的应用。

Design and heterologous expression of dengue virus envelope protein (E) peptides and their use for serological diagnosis.

机构信息

Research Center for Tropical Medicine - CEPEM, Porto Velho (RO), Brazil.

出版信息

J Virol Methods. 2012 Dec;186(1-2):55-61. doi: 10.1016/j.jviromet.2012.08.006. Epub 2012 Sep 7.

DOI:10.1016/j.jviromet.2012.08.006
PMID:22981980
Abstract

Viruses belonging to the Flaviviridae family are found and distributed in most of the tropical and sub-tropical regions of the world. The genus has more than 56 members, most of which cause clinical symptoms in humans. The clinical diagnosis of dengue requires laboratory confirmation because of the similarity of symptoms with a series of other acute fevers and the primary use antibodies or antigens for detection. In this work, peptides E(1) and E(2) of the envelope protein (E) of the dengue virus were mapped using bioinformatics methods. These peptides were then expressed in a prokaryotic system and purified. An indirect ELISA for antibodies IgG and IgM from laboratory samples previously characterised was then used with the peptides to detect anti-dengue antibodies. For IgG using the peptide E(1), the sensitivity of the indirect ELISA was 88.3% and the specificity was 56%; using the peptide E(2), the sensitivity was 90% and the specificity was 59%; and using a combination of both peptides, the sensitivity was 93.3% and the specificity was 78%. For IgM using the peptide E(1), the sensitivity was 88% and the specificity was 66%; using the peptide E(2), the sensitivity was 88% and the specificity was 69%; and when used in combination, the peptides E(1)/E(2) demonstrated a sensitivity of 90% and a specificity of 86%. These results indicate that the use of the E(1) and E(2) peptides of the E protein are an alternative for serological diagnosis of dengue fever.

摘要

属于黄病毒科的病毒在世界上大多数热带和亚热带地区都有发现和分布。该属有超过 56 个成员,其中大多数会在人类中引起临床症状。由于症状与一系列其他急性发热相似,且主要使用抗体或抗原进行检测,因此登革热的临床诊断需要实验室确认。在这项工作中,使用生物信息学方法对登革热病毒包膜蛋白(E)的 E1 和 E2 肽进行了定位。然后在原核系统中表达这些肽并进行纯化。然后使用间接 ELISA 检测先前特征化的实验室样本中的 IgG 和 IgM 抗体与肽的结合情况,以检测抗登革热抗体。对于使用肽 E1 的 IgG,间接 ELISA 的灵敏度为 88.3%,特异性为 56%;使用肽 E2 的灵敏度为 90%,特异性为 59%;使用两种肽的组合,灵敏度为 93.3%,特异性为 78%。对于使用肽 E1 的 IgM,灵敏度为 88%,特异性为 66%;使用肽 E2 的灵敏度为 88%,特异性为 69%;当两种肽联合使用时,E1/E2 肽的灵敏度为 90%,特异性为 86%。这些结果表明,E 蛋白的 E1 和 E2 肽可用于登革热的血清学诊断。

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