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Implementing CLIA for QuantiFERON®-CMV testing: Optimizing cellular immune response against cytomegalovirus
Juliana Esperalba1, Maria Arnedo-Muñoz2, Ariadna Rando-Segura3
1Microbiology Department, Vall d'Hebron Hospital Universitari, Vall d'Hebron Institut de Recerca (VHIR), Vall d'Hebron Barcelona Hospital Campus, Barcelona, Spain; Genetics and Microbiology Department, Universitat Autònoma de Barcelona, Bellaterra, Spain; Centro de Investigación Biomédica en Red de Enfermedades Infecciosas (CIBERINFEC), Instituto de Salud Carlos III, Madrid, Spain.
Insights
Chemiluminescence immunoassay (CLIA) offers a reliable alternative to ELISA for Cytomegalovirus (CMV) interferon-gamma release assays in transplant recipients. This method demonstrates excellent agreement and may improve laboratory efficiency.
Area of Science:
- Immunology
- Transplant Medicine
- Infectious Diseases
Background:
- Cytomegalovirus (CMV) infections are a major concern for solid organ transplant recipients.
- Interferon-gamma (IFN-γ) release assays, like QuantiFERON®-CMV (QTF-CMV), are crucial for managing CMV cellular immunity.
- Chemiluminescence immunoassay (CLIA) presents potential technical advantages over the standard ELISA method for QTF-CMV.
Purpose of the Study:
- To evaluate the performance of a CLIA-based assay (LIAISON® QuantiFERON®-TB Gold Plus) for quantifying IFN-γ in QTF-CMV testing.
- To compare the CLIA method directly against the established ELISA-based QuantiFERON®-CMV assay.
Main Methods:
- Retrospective analysis of 169 plasma samples from 85 lung transplant recipients.
- Quantification of IFN-γ using both ELISA (Qiagen®) and CLIA (DiaSorin®) platforms.
- Statistical comparison included correlation coefficients, Bland-Altman analysis, ROC curves, Kappa index, sensitivity, and specificity.
Main Results:
- CLIA produced significantly higher IFN-γ levels compared to ELISA for both specific (CMV-NIL) and nonspecific (MIT-NIL) responses.
- High correlation (r and ρc >0.93) and diagnostic agreement (95.86%, Kappa=0.913) were observed between the two methods.
- CLIA showed fewer indeterminate results (3.55% vs 4.73%) and achieved 100% sensitivity and specificity with an optimal cut-off of 0.2754 IU/mL.
Conclusions:
- The CLIA method demonstrates excellent agreement with ELISA for QTF-CMV testing, establishing it as a reliable alternative.
- CLIA offers potential for improved laboratory workflows due to its automatable nature.
- Adjusting the cut-off value is recommended for optimal diagnostic accuracy when using CLIA for QTF-CMV.
Introduction:
Cytomegalovirus (CMV) remains a significant infectious complication in solid organ transplant recipients. Assessing cellular immunity through interferon-gamma (IFN-γ) release assays, such as QuantiFERON®-CMV (QTF-CMV), is useful for guiding prophylactic and therapeutic decisions. Although ELISA is the validated method for QTF-CMV, chemiluminescence immunoassay (CLIA) may offer technical advantages. This study aimed to evaluate the performance of the CLIA-based LIAISON® QuantiFERON®-TB Gold Plus assay in quantifying IFN-γ in QTF-CMV testing, compared to the ELISA-based QuantiFERON®-CMV assay.
Methods:
A retrospective study was conducted on 169 stored plasma samples from 85 lung transplant recipients. IFN-γ was quantified using both ELISA (Qiagen®) and CLIA (DiaSorin®). Specific (CMV-NIL) and nonspecific (MIT-NIL) responses were compared using Pearson and Lin's concordance correlation coefficients, Bland-Altman analysis, and ROC curves. Diagnostic agreement was assessed using the Kappa index, sensitivity, specificity, and agreement rates.
Results:
CLIA yielded significantly higher IFN-γ values than ELISA for CMV-NIL (+0.41IU/mL) and MIT-NIL (+0.33IU/mL). Strong correlation and concordance were observed (r and ρc >0.93). Diagnostic agreement reached 95.86% (Kappa=0.913), with fewer indeterminate results in CLIA (3.55% vs 4.73%). The optimal CLIA cut-off was 0.2754IU/mL, achieving 100% sensitivity and specificity compared to ELISA.
Conclusions:
CLIA shows excellent agreement with ELISA in QTF-CMV testing and is a reliable and automatable alternative. Its implementation may improve laboratory workflows. Adjusting the cut-off is recommended for optimal diagnostic accuracy.
