Premium
Fetal erythroblasts from maternal blood identified with 2,3‐bisphosphoglycerate (BPG) and in situ hybridization (ISH) using Y‐specific probes
Author(s) -
Von Koskull Harriet,
Gahmberg Nina
Publication year - 1995
Publication title -
prenatal diagnosis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.956
H-Index - 97
eISSN - 1097-0223
pISSN - 0197-3851
DOI - 10.1002/pd.1970150207
Subject(s) - in situ hybridization , fetus , biology , obstetrics , andrology , medicine , genetics , pregnancy , gene , gene expression
Different types of fetal nucleated cells can be found in maternal blood, providing the possibility of non‐invasive prenatal diagnosis. For this purpose, we have studied fetal erythroblasts. We discovered that haemoglobin‐containing cells treated with 2,3‐bisphosphoglycerate (BPG) can be visualized by a peroxidase reaction, which at the same time visualizes an in situ hybridization (ISH) signal, specific for the X, Y or 21 chromosome. In order to prove that the BPG‐positive cells were erythroid, an anti‐glycophorin A (GPA) antiserum combined with a staphylococcal rosette technique was used. To enrich for erythroblasts, leukocytes were depleted from maternal blood by treatment with anti‐CD45 monoclonal antibody and passage over an anti‐mouse IgG‐coated glass bead column. To evaluate the potential of the method for clinical use, we studied maternal blood samples from 18 women referred to us for prenatal diagnosis between 6 and 19 weeks of gestation. Erythroblasts were found in 13 out of 14 normal pregnancies. Erythroblasts with a Y‐signal were found as early as 9 weeks of gestation, but at 6 weeks the Y‐signal was seen in BPG‐negative cells only. These cells showed an epithelioid morphology indicating that they were cytotrophoblasts. The BPG‐ISH method provides a simple technique for identifying erythroblasts and simultaneously visualizing a desired probe.