title: Comparison of commercial kits to measure cytokine responses to Plasmodium falciparum by multiplex microsphere suspension array technology.
abstract: BACKGROUND
  Multiplex cytokine profiling systems are useful tools for investigating correlates of protective immunity. Several Luminex and flow cytometry methods are commercially available but there is limited information on the relative performance of different kits. A series of comparison experiments were carried out to determine the most appropriate method for our subsequent studies.
  
  
  METHODS
  Two Luminex methods were compared, the Bio-Rad human 17-plex panel and the Invitrogen (formerly BioSource) human cytokine 10-plex kit, and two flow cytometry methods, the Becton Dickinson Human Th1/Th2 Cytokine Kit (CBA) and the Bender MedSystems Human Th1/Th2 11plex FlowCytomix Multiplex Kit. All kits were tested for the measurement of cytokines in supernatants collected from human leukocytes stimulated with viable Plasmodium falciparum infected red blood cells (iRBC) or P. falciparum schizont lysates.
  
  
  RESULTS
  Data indicated that the kits differed in sensitivity and reproducibility depending on the cytokine, and detected different quantities of some cytokines. The Bio-Rad 17-plex kit was able to detect more positive responses than the Invitrogen 10-plex kit. However, only when detecting IL-1, IL-6 or TNF did the two Luminex based methods correlate with one another. In this study, the flow cytometry based techniques were less variable and correlated better with one another. The two flow cytometry based kits showed significant correlation when detecting IFN-γ, IL-2, TNF, IL-10 and IL-6, but overall the BD kit detected more positive responses than the Bender MedSystems kit.
  
  
  CONCLUSIONS
  The microsphere suspension array technologies tested differed in reproducibility and the absolute quantity of cytokine detected. Sample volume, the number of cytokines measured, and the time and cost of the assays also differed. These data provide an accurate assessment of the four techniques, which will allow individual researchers to select the tool most suited for their study population.
authors.affiliation: Barcelona Centre for International Health Research, Hospital Clínic, Universitat de Barcelona, Barcelona, Spain.
authors.forename: Tamara K
authors.initials: TK
authors.lastname: Berthoud
authors.forename: Maria Nelia
authors.initials: MN
authors.lastname: Manaca
authors.forename: Diana
authors.initials: D
authors.lastname: Quelhas
authors.forename: Ruth
authors.initials: R
authors.lastname: Aguilar
authors.forename: Caterina
authors.initials: C
authors.lastname: Guinovart
authors.forename: Laura
authors.initials: L
authors.lastname: Puyol
authors.forename: Arnoldo
authors.initials: A
authors.lastname: Barbosa
authors.forename: Pedro L
authors.initials: PL
authors.lastname: Alonso
authors.forename: Carlota
authors.initials: C
authors.lastname: Dobaño
mesh_terms: D000328:Adult; D002478:Cells, Cultured; D019411:Clinical Laboratory Techniques / Q000379:methods*; D016207:Cytokines / Q000032:analysis*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology; D046228:Microarray Analysis / Q000379:methods*; D008863:Microspheres; D010963:Plasmodium falciparum / Q000276:immunology*; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D016207:Cytokines; D011933:Reagent Kits, Diagnostic