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通过补体结合试验和免疫扩散法对与传染性胃肠炎冠状病毒相关的主要可溶性抗原进行部分特性鉴定。

Partial characterization of the principal soluble antigens associated with the coronavirus of transmissible gastroenteritis by complement fixation and immunodiffusion.

作者信息

Stone S S, Kemeny L J, Jensen M T

出版信息

Infect Immun. 1976 Feb;13(2):521-6. doi: 10.1128/iai.13.2.521-526.1976.

Abstract

A microtiter complement fixation (CF) test to detect transmissible gastroenteritis (TGE) viral antigen was developed, using TGE hyperimmune pig serum as an antibody source. Sera from TGE covalescent pigs did not fix complement by this test. Maximal virus and soluble antigen (SA) titers were obtained 36 to 48 h after inoculation of swine testes cells. Cell-associated virus and SA titers were higher than those in the culture fluid, which had to be concentrated 20X before use as antigen in agar immunodiffusion tests (ID). By sucrose density-gradient centrifugation, the SA had a buoyant density of 1.10 g/ml and could be separated from the virus that banded in the 1.19-g/ml region. Virus and SA from three different isolates of TGE had the same buoyant densities. Heating and proteolytic enzyme digestion established the protein nature of the SA. As assayed by CF and ID, there were stability differences between crude and purified preparations of SA. Antibody prepared in rabbits against the SA neutralized the TGE virus.

摘要

利用传染性胃肠炎(TGE)超免疫猪血清作为抗体来源,开发了一种用于检测TGE病毒抗原的微量滴定补体结合(CF)试验。TGE康复猪的血清通过该试验不结合补体。在接种猪睾丸细胞36至48小时后获得最大病毒和可溶性抗原(SA)滴度。细胞相关病毒和SA滴度高于培养液中的滴度,培养液在用作琼脂免疫扩散试验(ID)的抗原之前必须浓缩20倍。通过蔗糖密度梯度离心,SA的浮力密度为1.10 g/ml,并且可以与在1.19-g/ml区域形成条带的病毒分离。来自三种不同TGE分离株的病毒和SA具有相同的浮力密度。加热和蛋白酶消化确定了SA的蛋白质性质。通过CF和ID测定,SA的粗制品和纯化制品之间存在稳定性差异。用兔制备的针对SA的抗体中和了TGE病毒。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d194/420642/c2de1d553665/iai00218-0221-a.jpg

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