title: Toward development of a comprehensive external quality assurance program for polyfunctional intracellular cytokine staining assays.
abstract: The External Quality Assurance Program Oversight Laboratory (EQAPOL) Flow Cytometry Program assesses the proficiency of NIH/NIAID/DAIDS-supported and potentially other interested research laboratories in performing Intracellular Cytokine Staining (ICS) assays. The goal of the EQAPOL Flow Cytometry External Quality Assurance Program (EQAP) is to provide proficiency testing and remediation for participating sites. The program is not punitive; rather, EQAPOL aims to help sites identify areas for improvement. EQAPOL utilizes a highly standardized ICS assay to minimize variability and readily identify those sites experiencing technical difficulties with their assays. Here, we report the results of External Proficiency 3 (EP3) where participating sites performed a 7-color ICS assay. On average, sites perform well in the Flow Cytometry EQAP (median score is "Good"). The most common technical issues identified by the program involve protocol adherence and data analysis; these areas have been the focus of site remediation. The EQAPOL Flow Cytometry team is now in the process of expanding the program to 8-color ICS assays. Evaluating polyfunctional ICS responses would align the program with assays currently being performed in support of HIV immune monitoring assays.
authors: Duke Center for AIDS Research, Duke University, Durham, NC, USA; Duke External Quality Assurance Program Oversight Laboratory, Duke University, Durham, NC, USA; Department of Surgery, Duke University Medical Center, Durham, NC, USA. Electronic address: janet.staats@dm.duke.edu.
authors: Janet S
authors: JS
authors: Staats
authors: Duke Center for AIDS Research, Duke University, Durham, NC, USA; Duke External Quality Assurance Program Oversight Laboratory, Duke University, Durham, NC, USA; Department of Surgery, Duke University Medical Center, Durham, NC, USA.
authors: Jennifer H
authors: JH
authors: Enzor
authors: Duke External Quality Assurance Program Oversight Laboratory, Duke University, Durham, NC, USA; Department of Medicine, Duke University Medical Center, Durham, NC, USA; Duke Human Vaccine Institute, Duke University, Durham, NC, USA.
authors: Ana M
authors: AM
authors: Sanchez
authors: Duke External Quality Assurance Program Oversight Laboratory, Duke University, Durham, NC, USA; Department of Medicine, Duke University Medical Center, Durham, NC, USA; Duke Human Vaccine Institute, Duke University, Durham, NC, USA.
authors: Wes
authors: W
authors: Rountree
authors: Duke Center for AIDS Research, Duke University, Durham, NC, USA; Duke External Quality Assurance Program Oversight Laboratory, Duke University, Durham, NC, USA; Department of Biostatistics and Bioinformatics, Duke University Medical Center, Durham, NC, USA.
authors: Cliburn
authors: C
authors: Chan
authors: BD Biosciences, San Jose, CA, USA.
authors: Maria
authors: M
authors: Jaimes
authors: Custom Technology Team, BD Biosciences, San Diego, CA, USA.
authors: Ray Chun-Fai
authors: RC
authors: Chan
authors: Custom Technology Team, BD Biosciences, San Diego, CA, USA.
authors: Amitabh
authors: A
authors: Gaur
authors: Duke External Quality Assurance Program Oversight Laboratory, Duke University, Durham, NC, USA; Department of Medicine, Duke University Medical Center, Durham, NC, USA; Duke Human Vaccine Institute, Duke University, Durham, NC, USA.
authors: Thomas N
authors: TN
authors: Denny
authors: Duke Center for AIDS Research, Duke University, Durham, NC, USA; Duke External Quality Assurance Program Oversight Laboratory, Duke University, Durham, NC, USA; Department of Surgery, Duke University Medical Center, Durham, NC, USA.
authors: Kent J
authors: KJ
authors: Weinhold
mesh_terms: D015415:Biomarkers / Q000032:analysis; D032921:Consensus; D003299:Cooperative Behavior; D016207:Cytokines / Q000032:analysis*; D005434:Flow Cytometry / Q000592:standards*; D019983:Guideline Adherence / Q000592:standards; D015658:HIV Infections / Q000175:diagnosis* / Q000276:immunology / Q000628:therapy; D006801:Humans; D007391:International Cooperation; D007753:Laboratories / Q000592:standards*; D059021:Laboratory Proficiency Testing / Q000592:standards*; D015166:Monitoring, Immunologic / Q000592:standards*; D015337:Multicenter Studies as Topic / 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; D058996:Quality Improvement; D019984:Quality Indicators, Health Care / Q000592:standards*; D015203:Reproducibility of Results; D013048:Specimen Handling / Q000592:standards
chemical_list: D015415:Biomarkers; D016207:Cytokines
keywords: Antigen specific; Flow cytometry; Grading; ICS; Proficiency testing; Site performance