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Flow Cytometry as a Diagnostic Tool in Primary and Secondary Immune Deficiencies
Jocelyn R Farmer1, Michelle DeLelys2
1Division of Rheumatology, Allergy and Immunology, Massachusetts General Hospital, COX 201, MGH, 55 Fruit Street, Boston, MA 02114, USA; Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, USA.
Insights
Flow cytometry is a powerful diagnostic tool for immune deficiencies. Proper assay design is crucial for accurate results in evaluating primary and secondary immune disorders.
Area of Science:
- Clinical immunology
- Diagnostic laboratory science
Background:
- Flow cytometry is essential for diagnosing immune deficiencies.
- Accurate diagnostic assays require careful methodological considerations.
Purpose of the Study:
- To outline the methodological considerations for flow cytometry assay design.
- To highlight the clinical applications of flow cytometry in immune deficiency evaluation.
Main Methods:
- Specimen collection, marker/fluorochrome selection, antibody titration.
- Instrumentation, compensation, gating, and reference range development.
- Cross-validation of flow cytometry assays for lymphocyte subsets.
Main Results:
- Commonly used analyses include lymphocyte and T-cell subsets, naive/memory cells, double-negative T-cells, and plasmablasts.
- Flow cytometry is directly applicable to severe primary immune deficiencies.
- It aids in diagnosis and therapeutic monitoring of secondary immune deficiencies.
Conclusions:
- Flow cytometry is a versatile and powerful tool for immune deficiency diagnostics.
- Standardized assay design and validation are critical for reliable results.
- Clinical utility spans both primary and secondary immune disorders.
Abstract:
Flow cytometry is an incredibly powerful diagnostic tool in the evaluation of primary and secondary immune deficiencies. Assay design and setup involves a methodological consideration of specimen collection, marker and fluorochrome selection, antibody titration, instrumentation, compensation, gating, reference range development, and cross validation. Commonly used analyses for lymphocytes are the lymphocyte subset, T-cell subset, B-cell and T-cell naive/memory, double-negative T-cell, and plasmablast panels. Flow cytometry has direct clinical applicability to the workup of severe forms of primary immune deficiency disorders and is used diagnostically and for therapeutic monitoring in the context of secondary immune deficiency disorders.
