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Updated: Jun 30, 2025

Enzyme-linked Immunospot Assay ELISPOT: Quantification of Th-1 Cellular Immune Responses Against Microbial Antigens
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Important Considerations for ELISpot Validation

Sylvia Janetzki1

  • 1ZellNet Consulting, Inc., Fort Lee, NJ, USA. sylvia@zellnet.com.

Methods in Molecular Biology (Clifton, N.J.)
|March 19, 2024
PubMed

The ELISpot assay has a solid place in the immune monitoring field for over 40 years. It is an assay that can assess the function of single immune cells in a straightforward and easy-to-learn approach. Its use in basic research, translational, and clinical work has been documented in countless publications. Harmonization guidelines and invaluable tools for optimal assay performance and evaluation exist. However, the validation of an established ELISpot protocol has been left to diverse opinions about how to interpret and tackle typical validation parameters. This chapter addresses important considerations for ELISpot validation, including the interpretations of validation parameters for a meaningful description of assay performance.

Keywords:
AccuracyELISpotELISpot validationLimit of DetectionPrecisionRobustnessSpecificity

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