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Chronic Granulomatous Disease: A Multicentre Study of DHR Flow Cytometry Results and Clinical Correlations in
Nahla Mohammed Saif1, Zarina Thasneem Zainudeen1, Intan Juliana Abd Hamid1
1Primary Immunodeficiency Diseases Group, Department of Clinical Medicine, Advanced Medical and Dental Institute, Universiti Sains Malaysia, Pulau Pinang, Malaysia.
Insights
Developing internal reference ranges for the Dihydrorhodamine (DHR) test is crucial for accurately diagnosing Chronic Granulomatous Disease (CGD). This study highlights inter-center variations, emphasizing the need for standardized DHR testing ranges.
Area of Science:
- Immunology
- Medical Diagnostics
- Genetics
Background:
- Chronic Granulomatous Disease (CGD) is an inherited immunodeficiency stemming from defects in NADPH oxidase genes.
- Neutrophil NADPH oxidase function is assessed via the Dihydrorhodamine (DHR) test for CGD diagnosis.
- Current DHR testing lacks standardized internal reference ranges, impacting diagnostic accuracy.
Purpose of the Study:
- To establish internal reference ranges for DHR testing by determining optimal cut-off values for oxidising cell percentages and Mean Fluorescence Intensity (MFI).
- To improve the accuracy and interpretation of DHR test results for diagnosing CGD.
- To investigate potential differences in DHR test parameters between two diagnostic centers.
Main Methods:
- Retrospective analysis of DHR test data from 107 individuals referred for testing at Institut Perubatan dan Pergigian Termaju (IPPT) and Hospital Universiti Sains Malaysia (HUSM).
- Logistic regression analysis to explore relationships between patient groups and DHR test parameters.
- Receiver Operating Characteristic (ROC) curve analysis to assess the diagnostic precision of the DHR test parameters.
Main Results:
- Optimal cut-off points for differentiating CGD patients from healthy individuals were determined for both centers.
- IPPT: fMLP% 0.26%, FMLP MFI 9.15, PMA% 58.70%, PMA MFI 50.00.
- HUSM: fMLP% 3.86%, FMLP MFI 999.50, PMA% 45.69%, PMA MFI 1130.50.
- All tested DHR parameters demonstrated significant analytical ability with high Area Under the Curve (AUC) values.
Conclusions:
- The study confirms the critical importance of establishing internal reference ranges for accurate CGD diagnosis using the DHR test.
- Significant differences in DHR test results and optimal cut-off points were observed between the two participating centers.
- These findings underscore the necessity for standardized reference ranges in laboratory diagnostics to ensure consistent and reliable CGD testing.
Background:
Chronic granulomatous disease (CGD) is an inherited primary immunodeficiency disease that results from a defect in one of the respiratory burst oxidases (NADPH oxidase) genes that form its components. The diagnostic laboratory assessment for CGD includes evaluation of NADPH oxidase function in neutrophils, using DHR test. The main objective of this project is to develop an internal reference range by determining the relative proportion of oxidising cells percentages and the mean fluorescence intensity (MFI) for complete interpretation of the test results and more accurate results.
Methods:
A retrospective analysis was conducted on 107 individuals referred for DHR testing from Institut Perubatan dan Pergigian Termaju (IPPT) and Hospital Universiti Sains Malaysia (HUSM). Purposive sampling was employed. Logistic regression was used to explore relationships between groups and test parameters, and receiver operating characteristic curve (ROC) analysis assessed test precision.
Results:
From our findings, the optimum cut-point for IPPT to differentiate CGD patients from healthy group with fMLP% was 0.26%, and FMLP MFI was 9.15, PMA% was 58.70%, and PMA MFI was 50.00. For the HUSM data, the best possible cut-point for fMLP% was 3.86%, FMLP MFI was 999.50, PMA% was 45.69%, and PMA MFI was 1130.50. The parameters showed a good analytical ability since all the area under the curve (AUC) values were significant (P > 0.5).
Conclusion:
This study confirmed the importance of developing an internal reference range for accurate diagnosis of CGD. This data showed a difference between two centres in terms of test results and cut-points, highlighting the need for standardised reference ranges in diagnostic testing.
