Detection of circulating fetal cells utilizing automated microscopy: potential for noninvasive prenatal diagnosis of chromosomal aneuploidies

Objective As fetal cells can be indisputably identified through detection of Y FISH signals, we utilized all automated microscopy system developed to identify and enumerate cells bearing X and Y FISH signals. We further investipted the potential offetal licnioglobin expression as a gender independent marker for automated identification of fetal cells. Method For FISH-based scanning, verified fetal cells were identified based oil the presence of a single X-signal and individual signals for each of the two Y FISH probes. For cell identification based on fetal hemoglobin expression, Putative fetal cells were verified based on the presence of signals for anti-gamma or anti-E globin antibody, and FISH signals for the X- and Y-chromosomes. Results Fetal cells were identified, by FISH-based scanning, in 28 of the 29 maternal samples from pregnancies with male fetuses. Simple density gradient centrifugation achieved a 3- to 5-fold increase in the number of fetal cells detected. Conclusion Automated microscopy identified fetal cells in both first and second trimester maternal blood samples. Although We Were unable to detect fetal erythroblasts ill numbers sufficient for clinical diagnosis, C the ability to reliably detect fetal cells by FISH-based scanning opens the possibility for prenatal detection of chromosomal aberrations utilizing circulating fetal cells. Copyright (c) 2008 John Wiley & Sons. Ltd.