Prenatal diagnosis with use of fetal cells isolated from maternal blood: Five-color fluorescent in situ hybridization analysis on flow-sorted cells for chromosomes X, Y, 13, 18, and 21

• Published in: American journal of obstetrics and gynecology Vol. 179; no. 1; pp. 203 - 209
• Main Authors: Bischoff, Farideh Z., Lewis, Dorothy E., Nguyen, Dianne D., Murrell, Sarah, Schober, Wendy, Scott b, Jeffrey, Simpson, Joe Leigh, Elias, Sherman
• Format: Journal Article
• Language: English
• Published: Philadelphia, PA Mosby, Inc 01-07-1998; Elsevier
• Subjects: Biological and medical sciences; Chromosome Aberrations - diagnosis; Chromosome Disorders; Chromosomes, Human, Pair 13; Chromosomes, Human, Pair 18; Chromosomes, Human, Pair 21; Down Syndrome - diagnosis; Down Syndrome - genetics; Female; fetal cells in maternal blood; Fetus - cytology; Flow Cytometry; Fluorescence in situ hybridization; Gynecology. Andrology. Obstetrics; Humans; In Situ Hybridization, Fluorescence - methods; Male; Management. Prenatal diagnosis; Maternal-Fetal Exchange - physiology; Medical sciences; Predictive Value of Tests; Pregnancy; Pregnancy. Fetus. Placenta; Prenatal Diagnosis - methods; X Chromosome; Y Chromosome; fetal cells in maternal blood; flow cytometry; Fluorescence in situ hybridization; Chromosomal aberration; Human; Flow cytometry; Molecular hybridization; In situ; Chromosome D13; Chromosome G21; Red blood cell; Aneuploidy; Y-Chromosome; Blood; Prenatal; Fetal diseases; Chromosome E18; Mother; Isolated cell; Genetics; Fetus; X-Chromosome; Diagnosis; Molecular biology
• ISSN: 0002-9378; 1097-6868
• DOI: 10.1016/S0002-9378(98)70273-X

OBJECTIVE: Currently, prenatal diagnosis of chromosome abnormalities requires invasive techniques such as amniocentesis and chorionic villus sampling that carry small but finite risks of fetal loss. A noninvasive approach is to isolate fetal cells from maternal blood by flow sorting followed by genetic interphase analysis with fluorescence in situ hybridization. Because the ratio of fetal to maternal cells is relatively low after flow sorting and to detect 90% to 95% of fetal aneuploidies associated with serious birth defects, a 5-color fluorescent in situ hybridization strategy is necessary for simultaneous detection of chromosomes X, Y, 13, 18, and 21 in all flow-sorted nuclei recovered from a specimen.
STUDY DESIGN: Fetal nucleated red blood cells were isolated from maternal blood in 40 cases (10.4 to 27.0 weeks’ gestation) by flow cytometry on the basis of positive selection of CD71
+ (transferrin receptor), CD45
–, and LDS751 staining. Each case was evaluated for 5-color fluorescent in situ hybridization efficiency by determining the percentage of flow-sorted nuclei containing 8 hybridization signals for chromosomes X, Y, 13, 18, and 21.
RESULTS: A total of 42,312 flow-sorted nuclei from maternal blood samples were analyzed. In 5 of 16 (31%) cases with a male fetus, 0.16% of nuclei scored were identified as fetal by the presence of 1 signal each for chromosomes X and Y. Fetal trisomy 21 nuclei were accurately detected in 2 cases with a female fetus, each of which was subsequently confirmed.
CONCLUSIONS: Five-color interphase fluorescent in situ hybridization analysis can be used to effectively analyze rare fetal aneuploid nuclei in enriched flow-sorted cells isolated from maternal blood. (Am J Obstet Gynecol 1998;179:203-9.)