Rapid, sensitive diagnosis of hemolytic anemia using antihemoglobin antibody in hypotonic solution.
We have developed a new and simple flow cytometric method to detect damaged red blood cells (RBCs) using anti-Hb in hypotonic solution. We studied a total of 200 patients, including 62 patients with schistocytosis, 8 postsplenectomy patients, and 108 healthy controls. Peripheral blood (2 microl) was stained with phycoerythrin-conjugated (PE) antihemoglobin antibody (anti-Hb) in 0.6% (w/v) NaCl solution, and analyzed by flow cytometry omitting the washing step. The proportion of RBCs stained by anti-Hb was 0.55% (SD +/-0.23%) in normal controls and was significantly higher in patients with schistocytosis (2.95+/-2.95%, p <0.001). Six of 108 blood samples from normal controls and 60 of 62 samples from schistocytosis patients showed > or =1.01% stained RBCs (ie, values > mean+2SD of normal controls). The number of schistocytes counted by microscopic examination correlated with the proportion of RBCs stained by anti-Hb (r = 0.637, p <0.001). The proportions of stained RBCs in blood samples with malaria, spherocytosis, and elliptocytosis were also significantly higher than in normal controls. However, the results in postsplenectomy and iron-deficiency anemia (IDA) patients were not significantly different from the normal controls; the number of schistocytes in postsplenectomy patients was not related to the proportion of RBCs stained by anti-Hb. Based on these findings, flow cytometry of damaged RBCs using anti-Hb in hypotonic solution is a simple, sensitive, and accurate method to detect active hemolysis.
Lee
Wonbae
W
Department of Clinical Pathology and Pediatrics, Catholic University Medical College, Seoul, Korea.
Kim
Yonggoo
Y
Lim
Jihyang
J
Kim
Myungshin
M
Lee
Eun Jung
EJ
Lee
Ahwon
A
Lee
Kyo Young
KY
Kang
Chang Suk
CS
Kim
So-Young
SY
Han
Kyungja
K
Pai
Soo Hwan
SH
D000743:Anemia, Hemolytic / Q000175:diagnosis*; D018798:Anemia, Iron-Deficiency / Q000097:blood; D000906:Antibodies; D004612:Elliptocytosis, Hereditary / Q000097:blood; D004912:Erythrocytes / Q000502:physiology / Q000648:ultrastructure; D004913:Erythrocytes, Abnormal / Q000502:physiology; D005434:Flow Cytometry; D006454:Hemoglobins / Q000276:immunology*; D006461:Hemolysis; D006801:Humans; D007038:Hypotonic Solutions; D008288:Malaria / Q000097:blood; D009996:Osmotic Fragility; D011184:Postoperative Period; D012016:Reference Values; D012680:Sensitivity and Specificity; D012965:Sodium Chloride; D013156:Splenectomy; D013194:Staining and Labeling; D013997:Time Factors
D000906:Antibodies; D006454:Hemoglobins; D007038:Hypotonic Solutions; D012965:Sodium Chloride