Flow cytometric detection of viruses

被引:203
作者
Brussaard, CPD
Marie, D
Bratbak, G
机构
[1] Univ Bergen, Dept Microbiol, N-5020 Bergen, Norway
[2] CNRS, Biol Stn, INSU, F-29682 Roscoff, France
[3] Univ Paris 06, F-29682 Roscoff, France
关键词
flow cytometry; nucleic acid specific green fluorescent dyes; SYBR Green I; viral genome sizes; viruses;
D O I
10.1016/S0166-0934(99)00167-6
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Representatives from several different virus families (Baculoviridae, Herpesviridae, Myoviridae, Phycodnaviridae, Picornaviridae, Podoviridae, Retroviridae, and Siphoviridae) were stained using a variety of highly fluorescent nucleic acid specific dyes (SYBR Green I, SYBR Green IT, OliGreen, PicoGreen) and examined using a standard flow cytometer equipped with a standard 15 mW argon-ion laser. The highest green fluorescence intensities were obtained using SYBR Green I. DNA viruses with genome sizes between 48.5 and 300 kb could easily be detected. The fluorescence signals of the small genome-sized RNA viruses (7.4-14.5 kb) were found at the limit of detection. No significant linear relationship could be found between genome size and the green fluorescence intensity of the SYBR Green I stained virus preparations. To our knowledge, this is the first report of detecting and discriminating between a wide range of different viruses directly using flow cytometry. This rapid and precise assay represents a new and promising tool in the field of virology. (C) 2000 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:175 / 182
页数:8
相关论文
共 19 条