Toward development of a comprehensive external quality assurance program for polyfunctional intracellular cytokine staining assays

• Published in: Journal of immunological methods 2014-07, Vol.409, p.44-53
• Main Authors: Staats, Janet S., Enzor, Jennifer H., Sanchez, Ana M., Rountree, Wes, Chan, Cliburn, Jaimes, Maria, Chan, Ray Chun-Fai, Gaur, Amitabh, Denny, Thomas N., Weinhold, Kent J.
• Format: Article
• Language: English
• Subjects: Antigen specific; Biomarkers - analysis; Consensus; Cooperative Behavior; cytokines; Cytokines - analysis; Flow cytometry; Flow Cytometry - standards; Grading; Guideline Adherence - standards; HIV Infections - diagnosis; HIV Infections - immunology; HIV Infections - therapy; Human immunodeficiency virus; Humans; ICS; International Cooperation; Laboratories - standards; Laboratory Proficiency Testing - standards; monitoring; Monitoring, Immunologic - standards; Multicenter Studies as Topic - standards; Observer Variation; Practice Guidelines as Topic - standards; Predictive Value of Tests; Proficiency testing; Program Development; Program Evaluation; Quality Control; Quality Improvement; Quality Indicators, Health Care - standards; remediation; Reproducibility of Results; Site performance; Specimen Handling - standards; staining
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/j.jim.2014.05.021

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.