(บทความวิจัย) การกำหนดช่วงอ้างอิงของการทดสอบ Dihydrorhodamine 123 โดยใช้เครื่อง BD FACSLyric ในประชากรไทย สำหรับการวินิจฉัยโรค Chronic Granulomatous Disease

Nichakarn Narkpradit; Wuttikrai Kongsankum; Jarupa Soongsathitanon

Authors

Nichakarn Narkpradit Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand
Wuttikrai Kongsankum Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand
Jarupa Soongsathitanon Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand

Keywords:

Dihydrorhodamine 123, Chronic granulomatous disease, DHR assay

Abstract

Chronic granulomatous disease (CGD) is a primary immunodeficiency caused by defects in the NADPH oxidase complex. This impairment inhibits the generation of reactive oxygen species (ROS) during the oxidative burst, a process essential for eliminating bacterial and fungal pathogens. The main clinical features of the disease are recurrent infections and granulomatous formation in various organs. The flow cytometric dihydrorhodamine 123 (DHR) assay is a standard method for evaluating oxidative burst activity and is widely utilized to support CGD diagnosis. This method is rapid, simple to perform, and highly sensitive. In the Cellular and Molecular Biology Laboratory, Department of Immunology, Faculty of Medicine Siriraj Hospital, the DHR assay has been used as a diagnostic test for CGD for decade, however, reference values have not yet been established or reported. This study aimed to establish reference values for DHR assay using the BD FACSLyric flow cytometry to support accurate laboratory interpretation. Data were collected from 285 healthy individuals between 2019 to 2023. The reference interval (2.5^th-97.5^th percentiles) for the stimulation index (SI) ranged from 105 to 304. The median fluorescence intensity (MFI) of unstimulated samples ranged from 12 to 39, while PMA-stimulated samples ranged from 2,631 to 6,664. The SI cut-off value was defined as 105, corresponding to the 2.5^th percentile. Using this cut-off, the DHR assay demonstrated a sensitivity of 100% and a specificity of 97.5%. Furthermore, abnormal DHR patterns have been observed in the patients, which effectively support the diagnosis of CGD.

Author Biographies

Wuttikrai Kongsankum, Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand

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Jarupa Soongsathitanon, Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand

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