High-throughput isolation of giant viruses using high-content screening

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作者
Rania Francis
Yusuke Ominami
Jacques Yaacoub Bou Khalil
Bernard La Scola
机构
[1] Institut Hospitalo-Universitaire Méditerranée-Infection,
[2] Aix-Marseille Université,undefined
[3] Institut de Recherche pour le Développement (IRD),undefined
[4] UMR Microbes Evolution Phylogeny and Infections (MEPHI),undefined
[5] Hitachi High-Technologies Corporation,undefined
[6] Nanotechnology Solutions Business Group,undefined
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摘要
The race to discover and isolate giant viruses began 15 years ago. Metagenomics is counterbalancing coculture, with the detection of giant virus genomes becoming faster as sequencing technologies develop. Since the discovery of giant viruses, many efforts have been made to improve methods for coculturing amebas and giant viruses, which remains the key engine of isolation of these microorganisms. However, these techniques still lack the proper tools for high-speed detection. In this paper, we present advances in the isolation of giant viruses. A new strategy was developed using a high-throughput microscope for real-time monitoring of cocultures using optimized algorithms targeting infected amebas. After validating the strategy, we adapted a new tabletop scanning electron microscope for high-speed identification of giant viruses directly from culture. The speed and isolation rate of this strategy has raised the coculture to almost the same level as sequencing techniques in terms of detection speed and sensitivity.
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