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采用酶联凝集素检测法(ELLA)检测和滴定流感 A 病毒神经氨酸酶抑制(NAI)抗体。

Detection and Titration of Influenza A Virus Neuraminidase Inhibiting (NAI) Antibodies Using an Enzyme-Linked Lectin Assay (ELLA).

机构信息

Virus and Prion Research Unit, National Animal Disease Center, United States Department of Agriculture, Agriculture Research Service, Ames, IA, USA.

出版信息

Methods Mol Biol. 2020;2123:335-344. doi: 10.1007/978-1-0716-0346-8_24.

Abstract

The neuraminidase (NA) of influenza A viruses (IAV) is a structurally and antigenically important envelope glycoprotein. There are eleven known subtypes of NA of which two, N1 and N2, circulate in swine. The sialidase activity of NA is required for the release of nascent virus particles from infected cell membranes and inhibition of NA enzymatic activity can significantly reduce virus titers and duration of infection. Efforts to improve IAV vaccine technology in humans have focused on the generation of neuraminidase inhibiting (NAI) antibodies and should be considered in swine as well. The enzyme-linked lectin assay (ELLA) conducted in 96-well plates has enabled high-throughput analysis of serum samples for NAI antibody titers. Through the use of reverse genetics, custom antigen panels and antisera can be generated to encompass the antigenically diverse population of NA that circulate in swine. The ELLA is a robust method to assess NAI antibody titers and characterize the antigenic difference between NA antigens.

摘要

流感病毒 A(IAV)的神经氨酸酶(NA)是一种结构和抗原上都很重要的包膜糖蛋白。有 11 种已知的 NA 亚型,其中两种,N1 和 N2,在猪中流行。NA 的唾液酸酶活性对于从受感染的细胞膜中释放新生病毒颗粒是必需的,并且抑制 NA 的酶活性可以显著降低病毒滴度和感染持续时间。为了改进人类的流感病毒疫苗技术,人们主要集中于产生神经氨酸酶抑制(NAI)抗体,在猪中也应该考虑这一点。在 96 孔板中进行的酶联凝集素测定(ELLA)已经能够实现高通量分析血清样本中的 NAI 抗体滴度。通过使用反向遗传学,可以生成定制的抗原面板和抗血清,以涵盖在猪中流行的具有不同抗原性的 NA 抗原群体。ELLA 是一种评估 NAI 抗体滴度和表征 NA 抗原之间抗原差异的强大方法。

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