Poly-FISH: a technique of repeated hybridizations that improves cytogenetic analysis of fetal cells in maternal blood

Zhen, Dong Kai; Wang, J. Y.; Falco, Vincent M.; Weber, William; Delli-Bovi, Laurent C.; Bianchi, Diana W.

doi:10.1002/(sici)1097-0223(199811)18:11<1181::aid-pd410>3.0.co;2-k

Cited by 37 publications

(17 citation statements)

References 17 publications

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“…Mononuclear cells were separated by 1.090 mg/ml density of Percoll solution (Pharmacia Biotech, Piscataway, NJ, USA) following protocols previously described (Zhen et al, 1998). Subsequent separation of fetal nucleated erythrocytes (NRBCs) was performed using a phycoerythrin (PE)-conjugated antibody to the gamma chain of fetal haemoglobin (Cortex Biochem, San Leandro, CA, USA) in protocols previously described for¯uorescence activated cell sorting (FACS) (Zhen et al, 1998;DeMaria et al, 1996). Gamma chain positive cells were separated by FACS and sorted onto microscope slides with a positive charge (Fisher Scienti®c,.…”

Section: Fetal Cell Separation and Identi®cationmentioning

confidence: 99%

“…The dry slides containing the candidate fetal cells were incubated in freshly prepared methanol : acetic acid (3 : 1) for 30 min at RT to ®x cells to slides. FISH analysis was performed using chromosomes X, Y, and/or 21-, and 18-speci®c probes in protocols previously described (Zhen et al, 1998). Chromosome-speci®c FISH signals on fetal NRBCs were visualized under a¯uorescence microscope (Zeiss Axioskop, Germany) and photographed using a CCD camera (Photometrics).…”

Section: Fetal Cell Separation and Identi®cationmentioning

confidence: 99%

“…Fetal cells in maternal blood are a source of fetal material for non-invasive prenatal diagnosis of fetal chromosome as well as DNA abnormalities (Simpson et al, 1995;Sekizawa et al, 1996;DeMaria et al, 1996;Cheung et al, 1996;Zhen et al, 1998;Bianchi et al, 1999). We have previously reported that, on average, approximately 19 nucleated fetal cells are detectable in 16 ml of maternal blood (Bianchi et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

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Apoptosis in fetal nucleated erythrocytes circulating in maternal blood

et al. 2000

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Section: Fetal Cell Separation and Identi®cationmentioning

confidence: 99%

Section: Fetal Cell Separation and Identi®cationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Apoptosis in fetal nucleated erythrocytes circulating in maternal blood

et al. 2000

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“…Fetal gender can be predicted by FISH analysis using X-and Y-specific probes. Their group also developed the 'poly-FISH' analysis, [34] which can be repeated at least 9 times on the same interphase nucleus and simultaneously uses at least 3 different probes. Theoretically, all 24 chromosomes can be analyzed on a single interphase fetal cell isolated from maternal blood.…”

Section: Identification Of Fetal Cells and Application For Prenatal Dmentioning

confidence: 76%

Prenatal Screening of Single-Gene Disorders from Maternal Blood

Sekizawa

Saitô

2001

American Journal of PharmacoGenomics

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Fetal cells and cell-free fetal DNA can be found circulating in maternal blood. Fetal cells recovered from maternal blood provide the only source of pure fetal DNA for noninvasive prenatal DNA diagnosis. Fetal nucleated erythrocytes (NRBCs) are considered the most suitable maternally-circulating fetal cells for this purpose, because they are not commonly found in the peripheral blood of healthy adults and are most abundant in the fetus during early gestation. Because fetal cells in maternal blood are extremely rare, a definitive separation method has not yet been established. Fetal NRBCs can be enriched from maternal blood via fluorescence- or magnetic-activated cell sorting, density gradients, immuno-magnetic beads or micromanipulation. Fetal cells are identified by Giemsa staining, hybridization with Y-chromosome specific probes, PCR-detection of a specific paternal allele, or immunostaining for fetal cell antigens. Amplification of fetal DNA sequences by primer extension preamplification and PCR has allowed prenatal screening for Duchenne muscular dystrophy and the fetal RhD blood type. Sequence-specific hybridization has been used to detect sickle cell anemia and beta-thalassemia prenatally in heterozygous carriers of these disorders. The use of cell-free fetal DNA in maternal plasma for the diagnosis of single-gene disorders is limited to disorders caused by a paternally inherited gene or a mutation that can be distinguished from the maternally inherited counterpart. At present, fetal gender can be determined from maternal plasma. When a pregnant woman is a heterzygous carrier of an X-linked disorder, the determination of fetal gender is clinically very informative for first-step screening to avoid invasive amniocentesis. The non-invasive prenatal diagnosis of genetic disorders should be applied to pregnant women with a definite risk for a specific single-gene disorder.

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“…La FISH con sondas específicas para los cromosomas X, Y, 21, 13 y 18 permite determinar el origen fetal de los eritroblastos aislados (en caso de feto varón) e identificar las aneuploidías más frecuentes 49,50 . Krabchi et al 51 .…”

Section: Aislamiento De Eritroblastos Fetales En Sangre Maternaunclassified