title: A decade of improvements in Mimiviridae and Marseilleviridae isolation from amoeba.
abstract: Since the isolation of the first giant virus, the Mimivirus, by T.J. Rowbotham in a cooling tower in Bradford, UK, and after its characterisation by our group in 2003, we have continued to develop novel strategies to isolate additional strains. By first focusing on cooling towers using our original time-consuming procedure, we were able to isolate a new lineage of giant virus called Marseillevirus and a new Mimivirus strain called Mamavirus. In the following years, we have accumulated the world's largest unique collection of giant viruses by improving the use of antibiotic combinations to avoid bacterial contamination of amoeba, developing strategies of preliminary screening of samples by molecular methods, and using a high-throughput isolation method developed by our group. Based on the inoculation of nearly 7,000 samples, our collection currently contains 43 strains of Mimiviridae (14 in lineage A, 6 in lineage B, and 23 in lineage C) and 17 strains of Marseilleviridae isolated from various environments, including 3 of human origin. This study details the procedures used to build this collection and paves the way for the high-throughput isolation of new isolates to improve the record of giant virus distribution in the environment and the determination of their pangenome.
authors.affiliation: Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes (URMITE), Aix-Marseille Université, Marseille, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Pagnier
authors.forename: Dorine-Gaelle Ikanga
authors.initials: DG
authors.lastname: Reteno
authors.forename: Hanene
authors.initials: H
authors.lastname: Saadi
authors.forename: Mondher
authors.initials: M
authors.lastname: Boughalmi
authors.forename: Morgan
authors.initials: M
authors.lastname: Gaia
authors.forename: Meriem
authors.initials: M
authors.lastname: Slimani
authors.forename: Tatsiana
authors.initials: T
authors.lastname: Ngounga
authors.forename: Meriem
authors.initials: M
authors.lastname: Bekliz
authors.forename: Philippe
authors.initials: P
authors.lastname: Colson
authors.forename: Didier
authors.initials: D
authors.lastname: Raoult
authors.forename: Bernard
authors.initials: B
authors.lastname: La Scola
mesh_terms: D000656:Amoeba / Q000821:virology*; D004267:DNA Viruses / Q000145:classification* / Q000302:isolation & purification*; D057166:High-Throughput Screening Assays / Q000379:methods; D013048:Specimen Handling / Q000379:methods; D014773:Virology / Q000379:methods*