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Noninvasive Determination of Fetal RhD Status Using Fetal DNA in Maternal Serum and PCR

Department of Obstetrics and Gynecology, Baylor College of Medicne, 6550 Fannin Street, Suite 708B, Houston, TX 77030 bischoff{at}bcm.tmc.edu

Dianne D. Nguyen, BS

Deborah Marquéz-Do, BS

Kenneth J. Moise, Jr, MD

Joe Leigh Simpson, MD

Sherman Elias, MD

Departments of Obstetrics and Gynecology and Molecular and Human Genetics, Baylor College of Medicine, Houston, Texts

Objective: Because prenatal testing of fetal RhD status by amniocentesis carries small yet finite risks to the fetus and mother, this study sought to determine whether fetal DNA in maternal serum could be used to detect fetal RhD status by polymerase chain reaction (PCR).

Methods: A retrospective analysis was made of frozen serum specimens from 20 sensitized RhD-negative pregnant women (ranging from 15.0 to 36.0 weeks' gestation) who were confirmed by serology at birth to have been carrying RhD-positive fetuses. Eleven serum specimens from RhD-negative individuals served as controls. DNA was isolated from serum and used in two PCR-based methods to detect a 99 base pair (bp) DNA fragment specific for the RhD gene and a 113 bp fragment specific for the RhCE gene as control.

Results: Overall, in 14 (70%) of 20 RhD-positive fetuses the 99 base pair RhD-specific PCR product was detected. There was no false positive detection among the 11 control serum specimens.

Conclusion: The results illustrate the ability to detect fetal RhD sequences in maternal serum of sensitized women. Moreover, the findings demonstrate that fetal single-gene disorders can be detected prenatally by using DNA isolated only from maternal serum.

Key Words: Fetal RhD • noninvasive prenatal diagnosis • maternal serum DNA • polymerase chain reaction

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