Evaluation of red cell membrane cytoskeletal disorders using a flow cytometric method in South iran.
OBJECTIVE
The diagnosis of hereditary red blood cell (RBC) membrane disorders, and in particular hereditary spherocytosis (HS) and Southeast Asian ovalocytosis (SAO), is based on clinical history, RBC morphology, and other conventional tests such as osmotic fragility. However, there are some milder cases of these disorders that are difficult to diagnose. The application of eosin-5'-maleimide (EMA) was evaluated for screening of RBC membrane defects along with some other anemias. We used EMA dye, which binds mostly to band 3 protein and to a lesser extent some other membrane proteins, for screening of some membrane defects such as HS.


METHODS
Fresh RBCs from hematologically normal controls and patients with HS, SAO, hereditary elliptocytosis, hereditary spherocytosis with pincered cells, severe iron deficiency, thalassemia minor, and autoimmune hemolytic anemia were stained with EMA dye and analyzed for mean fluorescent intensity (MFI) using a flow cytometer.


RESULTS
RBCs from patients with HS and iron deficiency showed a significant reduction in MFI compared to those from normal controls (p<0.0001 and p<0.001, respectively), while macrocytic RBCs showed a significant increase in MFI (p<0.01). A significant correlation was shown between mean corpuscular volume and MFI, with the exceptions of HS and thalassemia minor.


CONCLUSIONS
Our results showed that the flow cytometric method could be a reliable diagnostic method for screening and confirmation, with higher sensitivity and specificity (95% and 93%, respectively) than conventional routine tests for HS patients prior to further specific membrane protein molecular tests.
Diagnostic Laboratory, Sciences and Research Technology Center, Shiraz University of Medical Sciences, Shiraz, Iran ; School of Para Medical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Habib Alah
HA
Golafshan
Diagnostic Laboratory, Sciences and Research Technology Center, Shiraz University of Medical Sciences, Shiraz, Iran ; School of Para Medical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Reza
R
Ranjbaran
Diagnostic Laboratory, Sciences and Research Technology Center, Shiraz University of Medical Sciences, Shiraz, Iran ; School of Para Medical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Tahereh
T
Kalantari
Diagnostic Laboratory, Sciences and Research Technology Center, Shiraz University of Medical Sciences, Shiraz, Iran ; School of Para Medical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Leili
L
Moezzi
Hematology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Mehran
M
Karimi
Diagnostic Laboratory, Sciences and Research Technology Center, Shiraz University of Medical Sciences, Shiraz, Iran ; School of Para Medical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Abbas
A
Behzad-Behbahani
Diagnostic Laboratory, Sciences and Research Technology Center, Shiraz University of Medical Sciences, Shiraz, Iran ; School of Para Medical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Farzaneh
F
Aboualizadeh
Diagnostic Laboratory, Sciences and Research Technology Center, Shiraz University of Medical Sciences, Shiraz, Iran ; School of Para Medical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Sedigheh
S
Sharifzadeh
Band 3; Flow cytometry; Membrane disorders; RBC