Isolation of fetal nucleated red blood cell from maternal blood using immunomagnetic beads for prenatal diagnosis.
The immunomagnetic beads method for isolation of fetal nucleated red blood cells (FNRBCs) from peripheral blood of 78 pregnant women for prenatal diagnosis was developed. The study subjects were classified into 8-10 and 11-14 weeks of gestation (n = 39 each). Peripheral blood cells were divided into two for the FNRBCs isolation using two protocols, one with anti-CD45 depletion followed by anti-CD71 and anti-GPA monoclonal antibodies and another without CD45 depletion. The use of CD45 depletion gave a slightly higher number of sorted cells but not significantly different (p > 0.05). The percentage of CD71+ and GPA+ cells obtained from 8-10 weeks and 11-14 weeks of gestation was not different (p > 0.05). The sensitivity in determining the sorted FNRBCs for male fetal sex by PCR using 8-10 and 11-14 weeks of gestation was generally 50 and 69%, respectively. The method so developed is simple and cost effective and may thus be applied for prenatal diagnosis.
Pongsritasana
T
T
Department of Obstetric and Gynecology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
Wongratanacheewin
S
S
Prasertcharoensuk
V
V
Sermswan
R W
RW
D015703:Antigens, CD / Q000378:metabolism; D004912:Erythrocytes; D005260:Female; D005333:Fetus; D005434:Flow Cytometry; D006021:Glycophorins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D018189:Immunomagnetic Separation; D017493:Leukocyte Common Antigens / Q000378:metabolism; D047589:Leukocyte Reduction Procedures; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D011990:Receptors, Transferrin / Q000378:metabolism; D012680:Sensitivity and Specificity; D012732:Sex Determination Analysis / Q000379:methods*
D015703:Antigens, CD; C068126:CD71 antigen; C409972:GYPA protein, human; D006021:Glycophorins; D011990:Receptors, Transferrin; D017493:Leukocyte Common Antigens