title: Propagation capacity of bacterial contaminants in platelet concentrates using a luciferase reporter system.
abstract: BACKGROUND
  Currently the use of molecular tools and techniques of Genetic Engineering in the study of microbial behavior in blood components has replaced the employment of classical methods of microbiology. This work focuses on the use of a novel lux reporter system for monitoring the contaminating propagation capacity of bacteria present in platelet concentrates under standard storage conditions in the blood bank.
  
  
  METHODS
  A miniTn5 promotor probe carrying the lux operon from Photorhabdus luminiscens (pUTminiTn5luxCDABEKm2) was used to construct four bacterial bioluminescent mutants: Escherichia coli, Salmonella typhi, Proteus mirabilis and Pseudomonas aeruginosa. Luminescent mutants were used for contamination tests with 20 CFU in platelet concentrates bags and were stored under standard storage conditions in the blood bank (100 rpm at 22 °C). The measurements of luminous activity and optical density were used to monitor bacterial proliferation during 7 days (168 h).
  
  
  RESULTS
  During the exponential growth phase (log) of bacterial strains, a lineal correlation between luminous activity vs biomass was observed (R(2) = 0.985, 0.976, 0.981) for E. coli::Tn5luxCDABEKm2, P. mirabilis::Tn5luxCDABEKm2 and P. auriginosa::Tn5luxCDABEKm2, respectively. The above indicates that metabolic activity (production of ATP) is directly related to biomass in this phase of microbial growth. While conducting experiments, the inability to propagate S. typhi::Tn5luxCDABEKm2 was detected. We can speculate that platelet concentrates contain specific components that prevent the propagation of S. typhi.
  
  
  CONCLUSIONS
  The use of luxCDABE system for the quantification of luminous activity is a rapid and sensitive alternative to study the propagation and auto-sterilization of bacterial contaminants in platelet concentrates.
authors: Departamento de Investigación, Desarrollo y Control de Calidad, Centro Nacional de la Transfusión Sanguínea, Av. Othón de Mendizábal 195, Col. Zacatenco, México D.F. C.P. 07360, Mexique.
authors: Juan Manuel
authors: JM
authors: Bello-López
authors: Departamento de Investigación, Desarrollo y Control de Calidad, Centro Nacional de la Transfusión Sanguínea, Av. Othón de Mendizábal 195, Col. Zacatenco, México D.F. C.P. 07360, Mexique.
authors: Gabriela
authors: G
authors: Ibáñez-Cervantes
authors: Departamento de Investigación, Desarrollo y Control de Calidad, Centro Nacional de la Transfusión Sanguínea, Av. Othón de Mendizábal 195, Col. Zacatenco, México D.F. C.P. 07360, Mexique.
authors: Verónica
authors: V
authors: Fernández-Sánchez
authors: Departamento de Investigación, Desarrollo y Control de Calidad, Centro Nacional de la Transfusión Sanguínea, Av. Othón de Mendizábal 195, Col. Zacatenco, México D.F. C.P. 07360, Mexique.
authors: José Antonio
authors: JA
authors: Arroyo-Pérez
authors: Departamento de Investigación, Desarrollo y Control de Calidad, Centro Nacional de la Transfusión Sanguínea, Av. Othón de Mendizábal 195, Col. Zacatenco, México D.F. C.P. 07360, Mexique. Electronic address: julieta.rojo@salud.gob.mx.
authors: Julieta
authors: J
authors: Rojo-Medina
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D001771:Blood Banks; D001792:Blood Platelets / Q000382:microbiology*; D001793:Blood Preservation / Q000379:methods*; D004251:DNA Transposable Elements; D004926:Escherichia coli; D017930:Genes, Reporter; D005818:Genetic Engineering / Q000379:methods; D006801:Humans; D008156:Luciferases / Q000235:genetics; D049449:Luminescence; D008163:Luminescent Measurements; D009154:Mutation; D020637:Photorhabdus; D017713:Platelet Transfusion / Q000295:instrumentation*; D016133:Polymerase Chain Reaction; D011513:Proteus mirabilis; D011550:Pseudomonas aeruginosa; D012485:Salmonella typhi
chemical_list: D004251:DNA Transposable Elements; D000255:Adenosine Triphosphate; D008156:Luciferases
keywords: Autosterilization; Contaminants propagation; Luciferase; Platelet concentrates