Abstract
To determine the frequency of fetal nucleated cells in maternal peripheral blood during different stages of pregnancy, 50 primigravidas were investigated by determining the frequency of cells with the Y chromosome using fluorescence in situ hybridization (FISH) of Y-specific repetitive sequences of the DYZ1 family. Polymerase chain reaction (PCR) amplifying the same part of the DYZ1 used as the probe in FISH and a single-copy Y-specific fragment was also carried out for genomic DNA from the same samples. Cells with the hybridization signal were detected by FISH at and after 15 weeks of pregnancy in all pregnant women who gave birth to boys. The ratio of cells with the signal to those without the signal ranged from 1 in 144,000 to 1 in 4,000 with a tendency to increase as the pregnancy advanced. The frequency of fetal cells estimated by the PCR experiments was significantly and positively correlated with that found by FISH. The present study suggests that fetal nucleated cells increase in maternal peripheral blood with advancing gestation, from less than 1 in 100,000 nucleated cells in the first trimester to around 1 in 10,000 at term. These frequencies were much lower than those reported by cytological methods.
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Adinolfi M, Camporese C, Carr T (1989) Gene amplification to detect fetal nucleated cells in pregnant women. Lancet II:328–329
Bianchi DW, Flint AF, Pizzimenti MF, Knoll JHM, Latt SA (1990) Isolation of fetal DNA from nucleated erythrocytes in maternal blood. Proc Natl Acad Sci USA 87:3279–3283
Ciaranfi A, Curchod A, Odartchenko N (1977) Survie de lymphocytes foetaux dans le sang maternel post-partum. Schweiz Med Wochenschr 107:134–138
Gänshirt-Ahlert D, Pohlschmidt M, Gal A, Miny P, Horst J, Holzgreve W (1990) Ratio of fetal to maternal DNA is less than 1 in 5000 at different gestational ages in maternal blood. Clin Genet 38:38–43
Grosset L, Barrelet V, Odartchenko N (1974) Antenatal fetal sex determination from maternal blood during early pregnancy. Am J Obstet Gynecol 120:60–63
Grouchy J de, Trebuchet C (1971) Transfusion foeto-maternelle de lymphocytes sanguins et detection du sexe du foetus. Ann Genet 14:133–137
Herzenberg LA, Bianchi DW, Schröder J, Cann HM, Iverson GM (1979) Fetal cells in the blood of pregnant women: detection and enrichment by fluorescence-activated cell sorting. Proc Natl Acad Sci USA 76:1453–1455
Iverson GM, Bianchi DW, Cann HM, Herzenberg LA (1981) Detection and isolation of fetal cells from maternal blood using the fluorescence-activated cell sorter (FACS). Prenat Diagn 1:61–73
Kao SM, Tang GC, Hsieh TT, Young KC, Wang HC, Pao CC (1992) Analysis of peripheral blood of pregnant women for the presence of fetal Y chromosome-specific ZFY gene deoxyribonucleic acid sequences. Am J Obstet Gynecol 166:1013–1019
Kirsch-Volders M, Lissens-van Assche E, Susanne C (1980) Increase in the amount of fetal lymphocytes in maternal blood during pregnancy. J Med Genet 17:267–272
Kogan SC, Doherty M, Gitschier J (1987) An improved method for prenatal diagnosis of genetic diseases by analysis of amplified DNA sequences: application to hemophilia A. N Engl J Med 317:985–990
Kulozik A, Pawlowitzki IH (1982) Fetal cells in the maternal circulation: detection by direct AFP-immunofluorescence. Hum Genet 62:221–224
Kwok S, Higuchi R (1989) Avoiding false positives with PCR. Nature 339:237–238
Lawler M, McCann SR, Conneally E, Humphries P (1989) Chimaerism following allogeneic bone marrow transplantation: detection of residual host cells using the polymerase chain reaction. Br J Haematol 73:205–210
Lo YMD, Patel P, Wainscoat JS, Sampietro M, Gillmer MDG, Fleming KA (1989) Prenatal sex determination by DNA amplification from maternal peripheral blood. Lancet II: 1363–1365
Lo YMD, Patel P, Sampietro M, Gillmer MDG, Fleming KA, Wainscoat JS (1990) Detection of single-copy fetal DNA sequence from maternal blood. Lancet I: 1463–1464
Nakagome Y, Nagafuchi S, Nakahori Y (1990) Prenatal sex determination. Lancet 1:291
Nakagome Y, Seki S, Nagafuchi S, Nakahori Y, Sato K (1991) Absence of fetal cells in maternal circulation at a level of 1 in 25,000. Am J Med Genet 40:506–508
Parks DR, Herzenberg LA (1982) Fetal cells from maternal blood: their selection and prospects for use in prenatal diagnosis. Methods Cell Biol 26:277–295
Pinkel D, Straume T, Gray JW (1986) Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization. Proc Natl Acad Sci USA 83:2934–2938
Price JO, Elias S, Wachtel SS, Klinger K, Dockter M, Tharapel A, Shulman LP, et al (1991) Prenatal diagnosis with fetal cells isolated from maternal blood by multiparameter flow cytometry. Am J Obstet Gynecol 165:1731–1737
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Schröder J (1975) Transplacental passage of blood cells. J Med Genet 12:230–242
Schröder J, Chapelle A de la (1972) Fetal lymphocytes in the maternal blood. Blood 39:153–162
Schröder J, Tiilikainen A, Chapelle A de la (1974) Fetal leukocytes in the maternal circulation after delivery. Transplantation 17:346–354
Schwinger E, Hillers M, Vosberg HP (1989) No identification of Y-chromosomal DNA in blood from pregnant women bearing a male fetus? Am J Hum Genet 45:A268
Siebers JW, Knauf I, Hillemanns HG, Vogel W (1975) Antenatal sex determination in blood from pregnant women. Humangenetik 28:273–280
Walknowska J, Conte FA, Grumbach MM (1969) Practical and theoretical implications of fetal/maternal lymphocyte transfer. Lancet I:1119–1122
Zilliacus R, Chapelle A de la, Schröder J, Tiilikainen A, Kohne E, Kleihauer E (1975) Transplacental passage of foetal blood cells. Scand J Haematol 15:333–338
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Hamada, H., Arinami, T., Kubo, T. et al. Fetal nucleated cells in maternal peripheral blood: frequency and relationship to gestational age. Hum Genet 91, 427–432 (1993). https://doi.org/10.1007/BF00217766
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DOI: https://doi.org/10.1007/BF00217766