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[粪便标本中轮状病毒分析的免疫荧光反应]

[Immunofluorescence reaction for rotavirus analysis in fecal specimens].

作者信息

Arista S, Di Stefano R, Giovannelli L, Sinatra A

出版信息

Quad Sclavo Diagn. 1983 Dec;19(4):499-506.

PMID:6330786
Abstract

Testing for rotavirus detection in faeces from infants with acute diarrhea can be carried out, with high sensitivity, by immunofluorescence on rhesus monkey kidney cell lines (MA-104). Preincubation of viral inoculum with trypsin enhances further the sensitivity. In the present study we tried to assess the optimal conditions for detection of viral antigens, by analyzing the expression of virus-specific proteins at different times of infection and the effects of trypsin presence during virus adsorption and replication. Maximal production of viral antigens was obtained by infecting the cells 4 hours after cell-seeding and incubating them for 18 hours afterwards. In experiments with simian rotavirus SA-11, pretreated with trypsin and then with a trypsin inhibitor, a marked reduction in the percentage of fluorescent cells was observed thus indicating a role of the enzyme both on the virus and on the cell layer.

摘要

通过在恒河猴肾细胞系(MA - 104)上进行免疫荧光检测,可以高灵敏度地对患有急性腹泻的婴儿粪便中的轮状病毒进行检测。病毒接种物与胰蛋白酶的预孵育可进一步提高灵敏度。在本研究中,我们试图通过分析感染不同时间病毒特异性蛋白的表达以及病毒吸附和复制过程中胰蛋白酶存在的影响,来评估检测病毒抗原的最佳条件。在细胞接种后4小时感染细胞并随后孵育18小时,可获得病毒抗原的最大产量。在用胰蛋白酶预处理然后用胰蛋白酶抑制剂处理的猿猴轮状病毒SA - 11实验中,观察到荧光细胞百分比显著降低,这表明该酶在病毒和细胞层上均起作用。

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