Related Experiment Video
Updated: May 1, 2026
Optimized Interferon-gamma ELISpot Assay to Measure T Cell Responses in the Guinea Pig Model after Vaccination
Published on: January 20, 2019
The External Quality Assurance Oversight Laboratory (EQAPOL) proficiency program for IFN-gamma enzyme-linked
Ana M Sanchez1, Wes Rountree1, Mark Berrong2
1Duke Human Vaccine Institute, Duke University Medical Center, Durham, NC, USA.
Insights
An external proficiency program was developed to assess laboratory performance in interferon-gamma enzyme-linked immunospot (IFN-γ ELISpot) assays. This program ensures reliable data for infectious disease and cancer clinical trials by evaluating accuracy and precision.
Area of Science:
- Immunology
- Clinical Diagnostics
- Biotechnology
Background:
- The interferon-gamma enzyme-linked immunospot (IFN-γ ELISpot) assay is crucial for measuring antigen-specific immune responses in clinical trials.
- Standardized data comparison across laboratories is essential for evaluating therapeutic and vaccine strategies.
Purpose of the Study:
- To develop and implement an external proficiency program for the IFN-γ ELISpot assay.
- To establish a standardized method for evaluating laboratory performance and data concordance.
Main Methods:
- The program assesses laboratories on five parameters: timeliness, sample handling, background response detection, accuracy to consensus, and precision.
- A scoring system awards points for each criterion, resulting in a numeric and adjectival performance rating.
Main Results:
- The program provides laboratories and sponsors with information on the concordance of their results with other participating labs.
- Performance evaluation is based on accuracy to the consensus mean for detecting antigen-specific responses.
Conclusions:
- This external proficiency program offers a framework for organizing and implementing longitudinal performance evaluation of laboratories.
- It aims to meet Good Clinical Laboratory Practice (GCLP) guidelines for laboratories conducting end-point IFN-γ ELISpot assays in clinical trials.
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.
More Related Videos
06:13Enzyme-linked Immunospot Assay ELISPOT: Quantification of Th-1 Cellular Immune Responses Against Microbial Antigens
Published on: November 23, 2010
15:57Application of Long-term cultured Interferon-γ Enzyme-linked Immunospot Assay for Assessing Effector and Memory T Cell Responses in Cattle
Published on: July 11, 2015
