title: The External Quality Assurance Oversight Laboratory (EQAPOL) proficiency program for IFN-gamma enzyme-linked immunospot (IFN-γ ELISpot) assay.
abstract: The interferon-gamma enzyme-linked immunospot (IFN-γ ELISpot) assay has been developed and used as an end-point assay in clinical trials for infectious diseases and cancer to detect the magnitude of antigen-specific immune responses. The ability to compare data generated by different laboratories across organizations is pivotal to understand the relative potency of different therapeutic and vaccine strategies. We developed an external proficiency program for the IFN-γ ELISpot assay that evaluates laboratory performance based on five parameters: timeliness for data reporting; ability to handle cellular samples; detection of background (non-specific) responses; accuracy to consensus of the results; and precision of the measurements. Points are awarded for each criterion, and the sum of the points is used to determine a numeric and adjectival performance rating. Importantly, the evaluation of the accuracy to the consensus mean for the detection of antigen-specific responses using laboratory-specific procedures informs each laboratory and its sponsor on the degree of concordance of its results with those obtained by other laboratories. This study will ultimately provide the scientific community with information on how to organize and implement an external proficiency program to evaluate longitudinally the performance of the participating laboratories and, therefore, fulfill the requirements of the GCLP guidelines for laboratories performing end-point IFN-γ ELISpot assay for clinical trials.
authors.affiliation: Duke Human Vaccine Institute, Duke University Medical Center, Durham, NC, USA.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Sanchez
authors.affiliation: Duke Human Vaccine Institute, Duke University Medical Center, Durham, NC, USA.
authors.forename: Wes
authors.initials: W
authors.lastname: Rountree
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, NC, USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Berrong
authors.affiliation: Duke Human Vaccine Institute, Duke University Medical Center, Durham, NC, USA.
authors.forename: Ambrosia
authors.initials: A
authors.lastname: Garcia
authors.affiliation: Department of Retrovirology, USAMC-AFRIMS, Bangkok, Thailand.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Schuetz
authors.affiliation: International AIDS Vaccine Initiative, New York, New York, USA.
authors.forename: Josephine
authors.initials: J
authors.lastname: Cox
authors.affiliation: Fred Hutchinson Cancer Research Center, Seattle, Washington, USA.
authors.forename: Nicole
authors.initials: N
authors.lastname: Frahm
authors.affiliation: Department of Diagnostics and Monitoring, US Military HIV Research Program (MHRP), HJF, Walter Reed Army Institute of Research, Silver Spring, MD, USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Manak
authors.affiliation: Duke Human Vaccine Institute, Duke University Medical Center, Durham, NC, USA.|Department of Surgery, Duke University Medical Center, Durham, NC, USA.|Department of Immunology, Duke University Medical Center, Durham, NC, USA.|Duke Center for AIDS Research, Duke University Medical Center, Durham, NC, USA.
authors.forename: Marcella
authors.initials: M
authors.lastname: Sarzotti-Kelsoe
authors.affiliation: Vaccine Clinical Research Branch, Division of AIDS, NIAID, Bethesda, MD, USA.
authors.forename: M Patricia
authors.initials: MP
authors.lastname: D'Souza
authors.affiliation: Duke Human Vaccine Institute, Duke University Medical Center, Durham, NC, USA.|Department of Medicine, Duke University Medical Center, Durham, NC, USA.|Duke Global Health Institute; Duke University Medical Center, Durham, NC, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Denny
authors.affiliation: Duke Human Vaccine Institute, Duke University Medical Center, Durham, NC, USA.|Department of Surgery, Duke University Medical Center, Durham, NC, USA.|Duke Center for AIDS Research, Duke University Medical Center, Durham, NC, USA.|Duke Global Health Institute; Duke University Medical Center, Durham, NC, USA.
authors.forename: Guido
authors.initials: G
authors.lastname: Ferrari
mesh_terms: D002986:Clinical Trials as Topic / Q000592:standards*; D032921:Consensus; D003299:Cooperative Behavior; D058501:Enzyme-Linked Immunospot Assay / Q000592:standards*; D019983:Guideline Adherence / Q000592:standards; D006801:Humans; D059425:Interferon-gamma Release Tests / Q000592:standards*; D007391:International Cooperation; D007753:Laboratories / Q000592:standards*; D059021:Laboratory Proficiency Testing / Q000592:standards*; D008137:Longitudinal Studies; D015166:Monitoring, Immunologic / Q000592:standards*; D015588:Observer Variation; D017410:Practice Guidelines as Topic / Q000592:standards; D011237:Predictive Value of Tests; D016730:Program Development; D015397:Program Evaluation; D011786:Quality Control; D019984:Quality Indicators, Health Care / Q000592:standards*; D015203:Reproducibility of Results; D013048:Specimen Handling / Q000592:standards; D013997:Time Factors
keywords: ELISpot; Proficiency testing; Standardization