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猪血清中甲型流感病毒的微量中和试验。

Microneutralization assay for swine influenza virus in swine serum.

作者信息

Kitikoon Pravina, Vincent Amy L

机构信息

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

出版信息

Methods Mol Biol. 2014;1161:325-35. doi: 10.1007/978-1-4939-0758-8_27.

Abstract

The microneutralization (MN) assay is a modification of the serum virus neutralization assay and is a serological test to detect the presence of functional systemic antibodies that prevent infectivity of virus. When infectious virus is mixed with serum antibody, the virus infectivity can be "neutralized" if the antibodies bind to blocking epitopes on the virus. The neutralization effect can be demonstrated by inoculation of susceptible cells or organisms with the antibody-virus mixture, such as cells in culture, embryonated eggs, or susceptible hosts. The results of the MN assay described here are measured based on cell culture in a microtiter plate format and a color change detected by an automated plate reader. The test is performed with a constant amount of virus and serial dilutions of serum samples to an end point where virus neutralization is no longer detected. The neutralizing antibody titer is thus the reciprocal number of the last dilution of serum with neutralizing activity. The MN assay can be used to detect antibody from pigs with natural exposure or vaccination and can potentially be used to predict cross-protection between strains of influenza A virus.

摘要

微量中和(MN)试验是血清病毒中和试验的一种改良方法,是一种血清学检测,用于检测能够阻止病毒感染性的功能性全身抗体的存在。当感染性病毒与血清抗体混合时,如果抗体与病毒上的阻断表位结合,病毒的感染性就可以被“中和”。通过用抗体 - 病毒混合物接种易感细胞或生物体,如培养中的细胞、鸡胚或易感宿主,可以证明中和效果。这里描述的MN试验结果是基于微量滴定板形式的细胞培养以及通过自动酶标仪检测到的颜色变化来测量的。该试验使用恒定数量的病毒和血清样本的系列稀释液进行,直至检测不到病毒中和的终点。因此,中和抗体滴度是具有中和活性的血清最后一次稀释的倒数。MN试验可用于检测自然感染或接种疫苗的猪的抗体,并有可能用于预测甲型流感病毒毒株之间的交叉保护作用。

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