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A flow cytometric method for phenotyping recipient red cells following transfusion
G D Griffin1, L E Lippert, N S Dow
1Department of Pathology, Walter Reed Army Medical Center, Washington, D.C.
Transfusion
|March 1, 1994
Summary
A new dual-color flow cytometry method simplifies reticulocyte phenotyping for transfused patients. This technique accurately identifies red blood cell antigens in mixed populations, improving blood transfusion matching.
Area of Science:
- Hematology
- Immunology
- Biotechnology
Background:
- Reticulocyte phenotyping is crucial for transfused patients with red cell antibodies to ensure compatible blood transfusions.
- Current methods are time-consuming, labor-intensive, and require large sample volumes.
Purpose of the Study:
- To develop and validate a simpler, more efficient method for antigen typing of reticulocytes in mixed red cell populations.
Main Methods:
- A dual-color flow cytometry technique using indirect immunofluorescence for antigen labeling and thiazole orange for reticulocyte staining.
- Validation involved analyzing 319 samples with simulated transfusion concentrations (25-75%) of mixed red blood cells.
Main Results:
- The method accurately distinguished antigen-positive and -negative reticulocyte populations across various simulated transfusion levels.
- Successful antigen typing was demonstrated for multiple red blood cell antigens (e.g., c, C, K, s, Fya, Fyb, Jka, Jkb), with anti-S yielding inconsistent results.
- In vitro findings were confirmed in 19 transfused patients and one chimera blood donor.
Conclusions:
- This novel flow cytometry method offers a simpler, safer, and less labor-intensive approach to reticulocyte antigen typing.
- It requires significantly less sample volume compared to existing techniques, making it more practical for clinical use.