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HelpTest Code DHR Dihydrorhodamine Flow Cytometric Test, Blood
Useful For
Evaluation of chronic granulomatous disease (CGD), X-linked and autosomal recessive forms, Rac2 deficiency, complete myeloperoxidase deficiency
Monitoring chimerism and nicotinamide adenine dinucleotide phosphate oxidase (NOX) function post-hematopoietic cell transplantation
Assessing residual NOX activity pretransplant
Identifying female carriers for X-linked CGD
Assessing changes in lyonization with age in female carriers
Reporting Name
DHR Flow, BSpecimen Type
WB Sodium HeparinShipping Instructions
Specimens received on days when testing is not performed or after 5 p.m. Central on Friday will be canceled if specimen is outside of stability when testing is next performed.
Collect and package specimen as close to shipping time as possible. Ship specimen overnight in an Ambient Shipping Box-Critical Specimens Only (T668) following the instructions in the box. It is recommended that specimens arrive within 24 hours of collection.
Necessary Information
Ordering healthcare professional name and phone number are required.
Specimen Required
Two whole-blood sodium heparin specimens are required, one from the testing patient and the other from an unrelated healthy donor as a control.
Supplies: Ambient Shipping Box-Critical Specimens Only (T668)
Patient:
Container/Tube: Green top (sodium heparin)
Specimen Volume: 5 mL
Collection Instructions: Send whole blood specimen in original tube. Do not aliquot.
Normal Control:
Container/Tube: Green top (sodium heparin)
Specimen Volume: 5 mL
Collection Instructions:
1. Collect a control specimen from the unrelated healthy donor within an hour of the patient's specimen collection time.
2. Clearly label as Normal Control on the outermost label.
3. Send the whole blood specimen in the original tube. Do not aliquot.
4. Rubber band patient specimen and control vial together.
Specimen Minimum Volume
1 mL
Specimen Stability Information
| Specimen Type | Temperature | Time | Special Container |
|---|---|---|---|
| WB Sodium Heparin | Ambient | 48 hours | GREEN TOP/HEP |
Reference Values
|
Result name |
Unit |
Cutoff for defining normal |
|
% PMA ox-DHR+ |
% |
≥95% |
|
MFI PMA ox-DHR+ |
MFI |
≥60 |
|
% fMLP ox-DHR+ |
% |
≥10% |
|
MFI fMLP ox-DHR+ |
MFI |
≥2 |
|
Control % PMA ox-DHR+ |
% |
≥95% |
|
Control MFI PMA ox-DHR+ |
MFI |
≥60 |
|
Control % fMLP ox-DHR+ |
% |
≥10% |
|
Control MFI fMLP ox-DHR+ |
MFI |
≥2 |
PMA = phorbol myristate acetate
DHR = dihydrorhodamine
MFI = mean fluorescence intensity
fMLP = N-formyl-methionyl-leucyl-phenylalanine
The appropriate age-related reference values for Absolute Neutrophil Count will be provided on the report.
Interpretation
An interpretive report will be provided, in addition to the quantitative values.
Interpretation of the results of the quantitative dihydrorhodamine (DHR) flow cytometric assay must include both the proportion of positive neutrophils for DHR after phorbol myristate acetate and/or N-formyl-methionyl-leucyl-phenylalanine stimulation and the mean fluorescence intensity. Additionally, visual assessment of the pattern of DHR fluorescence is helpful in discriminating between the various genetic defects associated with chronic granulomatous disease and complete myeloperoxidase deficiency.
Clinical Reference
1. Kang EM, Marciano BE, DeRavin SS, Zarember KA, Holland SM, Malech HL. Chronic granulomatous disease: Overview and hematopoietic stem cell transplantation. J Allergy Clin Immunol. 2011;127(6):1319-1328
2. Segal BH, DeCarlo ES, Kwon-Chung KJ, Malech HL, Gallin JI, Holland SM. Aspergillus nidulans infection in chronic granulomatous disease. Medicine (Baltimore). 1998;77(5):345-354
3. Arnadottir GA, Norddahl GL, Gudmundsdottir S, et al. A homozygous loss-of-function mutation leading to CYBC1 deficiency causes chronic granulomatous disease. Nat Commun. 2018;9(1):4447
4. van de Geer A, Nieto-Patlan A, Kuhns DB, et al. Inherited p40phox deficiency differs from classic chronic granulomatous disease. J Clin Invest. 2018;128(9):3957-3975. doi:10.1172/JCI97116
5. Kuhns DB, Alvord WG, Heller T, et al. Residual NADPH oxidase and survival in chronic granulomatous disease. N Engl J Med. 2010;363(27):2600-2610
6. Vowells SJ, Fleisher TA, Sekhsaria S, Alling DW, Maguire TE, Malech HL. Genotype-dependent variability in flow cytometric evaluation of reduced nicotinamide adenine dinucleotide phosphate oxidase function in patients with chronic granulomatous disease. J Pediatr. 1996;128(1):104-107
7. Vowells SJ, Sekhsaria S, Malech HL, Shalit M, Fleisher TA. Flow cytometric analysis of the granulocyte respiratory burst: a comparison study of fluorescent probes. J Immunol Methods. 1995;178(1):89-97
8. Mauch L, Lun A, O'Gorman MR, et al. Chronic granulomatous disease (CGD) and complete myeloperoxidase deficiency both yield strongly reduced dihydrorhodamine 123 test signals but can be easily discerned in routine testing for CGD. Clin Chem. 2007;53(5):890-896
9. Ambruso DR, Knall C, Abell AN, et al. Human neutrophil immunodeficiency syndrome is associated with an inhibitory Rac2 mutation. Proc Natl Acad Sci U S A. 2000;97(9):4654-4659
10. Accetta D, Syverson G, Bonacci B, et al. Human phagocyte defect caused by a Rac2 mutation detected by means of neonatal screening for T-cell lymphopenia. J Allergy Clin Immunol. 2011;127(2):535-538
11. Donko A, Sharapova SO, Kabat J, et al. Clinical and functional spectrum of RAC2-related immunodeficiency. Blood. 2024;143(15):1476-1487
12. Hsu AP, Donko A, Arrington ME, et al. Dominant activating RAC2 mutation with lymphopenia, immunodeficiency, and cytoskeletal defects. Blood. 2019;133(18):1977-1988
13. Roesler J. Carriers of X-linked chronic granulomatous disease at risk. Clin Immunol. 2009;130(2):233-234. doi:10.1016/j.clim.2008.09.013
14. Rosen-Wolff A, Soldan W, Heyne K, Bickhardt J, Gahr M, Roesler J. Increased susceptibility of a carrier of X-linked chronic granulomatous disease (CGD) to Aspergillus fumigatus infection associated with age-related skewing of lyonization. Ann Hematol. 2001;80(2):113-115
15. Yamada M, Okura Y, Suzuki Y, et al. Somatic mosaicism in two unrelated patients with X-linked chronic granulomatous disease characterized by the presence of a small population of normal cells. Gene. 2012;497(1):110-115
16. de Boer M, Bakker E, Van Lierde S, Roos D. Somatic triple mosaicism in a carrier of X-linked chronic granulomatous disease. Blood. 1998;91(1):252-257
17. Noack D, Heyworth PG, Kyono W, Cross AR. A second case of somatic triple mosaicism in the CYBB gene causing chronic granulomatous disease. Hum Genet. 2001;109(2):234-238
18. Wolach B, Scharf Y, Gavrieli R, de Boer M, Roos D. Unusual late presentation of X-linked chronic granulomatous disease in an adult female with a somatic mosaic for a novel mutation in CYBB. Blood. 2005;105(1):61-66
19. Siler U, Romao S, Tejera E, et al. Severe glucose-6-phosphate dehydrogenase deficiency leads to susceptibility to infection and absent NETosis. J Allergy Clin Immunol. 2017;139(1):212-219
20. Kuhns DB. Diagnostic testing for chronic granulomatous disease. Methods Mol Biol. 2019;1982:543-571
21. Delmonte OM, Fleisher TA. Flow cytometry: Surface markers and beyond. J Allergy Clin Immunol. 2019;143(2):528-537
22. Knight V, Heimall JR, Chong H, et al. A toolkit and framework for optimal laboratory evaluation of individuals with suspected primary immunodeficiency. J Allergy Clin Immunol Pract. 2021;9(9):3293-3307.e6
Day(s) Performed
Monday through Friday
Report Available
3 to 4 daysPerforming Laboratory
Mayo Clinic Laboratories in Rochester
CPT Code Information
86352 x2
LOINC Code Information
| Test ID | Test Order Name | Order LOINC Value |
|---|---|---|
| DHR | DHR Flow, B | 98122-5 |
| Result ID | Test Result Name | Result LOINC Value |
|---|---|---|
| ANC | Absolute Neutrophil Count | 751-8 |
| PMAP | % PMA ox-DHR+ | 85376-2 |
| PMAM | MFI PMA ox-DHR+ | 85374-7 |
| FMPPP | % FMLP ox-DHR+ | 85373-9 |
| FMPM | MFI fMLP ox-DHR+ | 85370-5 |
| ANCC | Control Absolute Neutrophil Count | 85369-7 |
| PMAPC | Control % PMA ox-DHR+ | 85377-0 |
| PMAMC | Control MFI PMA ox-DHR+ | 85375-4 |
| FMPPC | Control % fMLP ox-DHR+ | 85372-1 |
| FMPMC | Control MFI fMLP ox-DHR+ | 85371-3 |
| DHRI | Interpretation | 69052-9 |
Method Name
Flow Cytometry