Analysis of HLA-DQ alpha sequences for prenatal diagnosis in single fetal cells from maternal blood.
We have extended a previously developed method that allows prenatal DNA diagnosis of female fetuses through the isolation of single nucleated erythrocytes from maternal blood by developing a method that can distinguish between maternal and fetal nucleated erythrocytes. Nucleated erythrocytes were separated by a density-gradient method and then collected by micromanipulation. Sex was determined after primer extension preamplification (PEP) of the entire genome of a single cell, and human leukocyte antigen (HLA)-DQ alpha type was determined after further amplification of this gene. The HLA-DQ alpha genotype of fetal erythrocytes in maternal blood samples and their corresponding paternal and maternal lymphocytes were successfully determined in all cases. The accuracy of the method was determined by using single nucleated erythrocytes from umbilical cord blood from five normal deliveries. This is the first demonstration that the fetal HLA-DQ alpha gene sequences can be identified in a small aliquot of a single nucleated erythrocyte in maternal blood. We believe that this method ushers in a new era in which the reliability and accuracy of noninvasive prenatal DNA diagnosis from maternal blood is markedly improved.
Sekizawa
A
A
Department of Obstetrics and Gynecology, School of Medicine, Showa University, Shinagawa, Tokyo, Japan.
Taguchi
A
A
Watanabe
A
A
Kimura
T
T
Saito
H
H
Yanaihara
T
T
Sato
T
T
D000483:Alleles; D004900:Erythroblasts / Q000737:chemistry*; D005260:Female; D005333:Fetus; D005838:Genotype; D006683:HLA-DQ Antigens / Q000097:blood* / Q000235:genetics*; D059848:HLA-DQ alpha-Chains; D006650:Histocompatibility Testing; D006801:Humans; D008297:Male; D008431:Maternal-Fetal Exchange / Q000235:genetics; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*
D006683:HLA-DQ Antigens; D059848:HLA-DQ alpha-Chains; C069050:HLA-DQA1 antigen