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Fetal Gender Determination and BclI Polymorphism Using Nucleated Erythrocytes in Maternal Blood

Jin Choe, Doyeong Hwang, Ki-Chul Kim and Young-Min Choi

Hamchoon Women's Clinic (JC,DH,K-CK) and Department of Obstetrics and Gynecology, College of Medicine (Y-MC), Seoul National University, Seoul, Korea

Correspondence to: Jin Choe, MD, 1621-7 Hamchoon Women's Clinic, Seocho-1-dong Seocho-ku, Seoul, Korea 137-878. E-mail: jchoe{at}hamchoon.com

This study demonstrated determination of fetal gender from nucleated red blood cells (NRBCs) in maternal blood and attempted to apply prenatal diagnosis of hemophilia A using BclI DNA polymorphism. Venous blood was drawn from 20 pregnant women, and NRBCs were recovered by magnetic activated cell sorting and anti-GPA (glycophorin A) immunostaining. After microdissector isolation of the NRBCs, primer extension preamplification (PEP) and nested PCR of the amelogenin gene were performed to determine fetal gender. We also performed PEP and nested PCR of BclI polymorphism to verify the validity of prenatal diagnosis of hemophilia A. DNA amplification was achieved in 107 cells (51.9%) and fetal gender determined with 65.0% accuracy. Unfortunately, we could not verify the validity within the scope of this study. However, in a larger number of cases that are informative in BclI polymorphism, we will be able to identify patients affected by hemophilia A using fetal NRBCs in maternal blood. (J Histochem Cytochem 53:323–327, 2005)

Key Words: nucleated erythrocyte • fetal gender • hemophilia A • micromanipulation • primer extension • preamplification • polymerase chain reaction

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