TY - JOUR T1 - Quality-controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials. JF - PLoS One Y1 - 2009 A1 - Merabishvili, Maya A1 - Pirnay, Jean-Paul A1 - Verbeken, Gilbert A1 - Chanishvili, Nina A1 - Tediashvili, Marina A1 - Lashkhi, Nino A1 - Glonti, Thea A1 - Krylov, Victor A1 - Jan Mast A1 - Van Parys, Luc A1 - Lavigne, Rob A1 - Volckaert, Guido A1 - Wesley Mattheus A1 - Verween, Gunther A1 - De Corte, Peter A1 - Rose, Thomas A1 - Jennes, Serge A1 - Zizi, Martin A1 - De Vos, Daniel A1 - Vaneechoutte, Mario KW - Bacteriophages KW - Burns KW - Clinical Trials as Topic KW - Genome, Viral KW - Humans KW - Proteome KW - Pseudomonas aeruginosa KW - Pseudomonas Infections KW - Staphylococcal Infections KW - Staphylococcus aureus KW - wound infection AB -

We describe the small-scale, laboratory-based, production and quality control of a cocktail, consisting of exclusively lytic bacteriophages, designed for the treatment of Pseudomonas aeruginosa and Staphylococcus aureus infections in burn wound patients. Based on successive selection rounds three bacteriophages were retained from an initial pool of 82 P. aeruginosa and 8 S. aureus bacteriophages, specific for prevalent P. aeruginosa and S. aureus strains in the Burn Centre of the Queen Astrid Military Hospital in Brussels, Belgium. This cocktail, consisting of P. aeruginosa phages 14/1 (Myoviridae) and PNM (Podoviridae) and S. aureus phage ISP (Myoviridae) was produced and purified of endotoxin. Quality control included Stability (shelf life), determination of pyrogenicity, sterility and cytotoxicity, confirmation of the absence of temperate bacteriophages and transmission electron microscopy-based confirmation of the presence of the expected virion morphologic particles as well as of their specific interaction with the target bacteria. Bacteriophage genome and proteome analysis confirmed the lytic nature of the bacteriophages, the absence of toxin-coding genes and showed that the selected phages 14/1, PNM and ISP are close relatives of respectively F8, phiKMV and phage G1. The bacteriophage cocktail is currently being evaluated in a pilot clinical study cleared by a leading Medical Ethical Committee.

VL - 4 CP - 3 M3 - 10.1371/journal.pone.0004944 ER -