Enrichment of fetal trophoblast cells from the maternal peripheral blood followed by detection of fetal deoxyribonucleic acid with a nested X/Y polymerase chain reaction


      OBJECTIVE: Fetal cells circulate in the maternal blood during early pregnancy. Because these cells are rare, noninvasive prenatal diagnosis from fetal cells can be achieved only after efficient enrichment procedures. Our aim was to develop a two-step enrichment procedure to isolate trophoblast cells from 20 ml of peripheral blood. STUDY DESIGN: Blood was obtained from pregnant women between 6 and 15 weeks of gestation, before invasive procedures were performed. After enrichment, the success of isolating fetal cells was determined by amplification of Y chromosome sequences. RESULTS: A highly specific X/Y polymerase chain reaction was established, sensitive enough to detect X and Y chromosome - specific sequences in one single cell and in one male cell among 100,000 female cells. Sex determination by polymerase chain reaction was compared with results from conventional karyotyping. The success rate was 91.7%. CONCLUSION: Enrichment of trophoblast cells from maternal blood as described here might be useful for early noninvasive prenatal diagnosis. (AM J OBSTET GYNECOL 1996;174:871-6.)


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        • Simpson JL
        • Elias S.
        Isolating fetal cells from maternal blood: advances in prenatal diagnosis through molecular technology.
        JAMA. 1993; 270: 2357-2361
        • Simpson JL
        • Elias S.
        Isolating fetal cells in maternal circulation for prenatal diagnosis.
        Prenat Diagn. 1994; 14: 1229-1242
        • Chueh J
        • Golbus MS.
        Prenatal diagnosis using fetal cells from the maternal circulation.
        West J Med. 1993; 159: 308-311
        • Senyei AE
        • Wassman ER.
        Fetal cells in the maternal circulation: technical considerations for practical application to prenatal diagnosis.
        Obstet Gynecol Clin North Am. 1993; 20: 583-598
        • Holzgreve W
        • Garritsen HSP
        • Ganshirt-Ahlert D.
        Fetal cells in the maternal circulation.
        J Reprod Med. 1992; 37: 410-418
        • Bianchi DW
        • Sylvester S
        • Zickwolf GK
        • DeMaria MA
        • Weil GJ
        • Geifman OH.
        Fetal stem cells persist in maternal blood for decades post-partum.
        Am J Hum Genet. 1993; 53: 251
        • Mueller UW
        • Hawes CS
        • Wright AE
        • et al.
        Isolation of fetal trophoblast cells from peripheral blood of pregnant women.
        Lancet. 1990; 336: 197-200
        • Hawes CS
        • Suskin HA
        • Petropoulos A
        • Latham SE
        • Mueller UW.
        A morphologic study of trophoblast isolated from peripheral blood of pregnant women.
        AM J OBSTET GYNECOL. 1994; 170: 1297-1300
        • Cacheux V
        • Milesi-Fluet C
        • Tachdjian G
        • et al.
        Detection of 47,XYY trophoblast fetal cells in maternal blood by fluorescence in situ hybridization after using immunomagnetic lymphocyte depletion and flow cytometry sorting.
        Fetal Diagn Ther. 1992; 7: 190-194
        • Price JO
        • Elias S
        • Wachtel SS
        • et al.
        Prenatal diagnosis with fetal cells isolated from maternal blood by multiparameter flow cytometry.
        AM J OBSTET GYNECOL. 1991; 165: 1731-1737
        • Bianchi DW
        • Mahr A
        • Zickwolf GK
        • Houseal TW
        • Flint AF
        • Klinger KW
        Detection of fetal cells with 47,XY,+21 karyotype in maternal peripheral blood.
        Hum Genet. 1992; 90: 368-370
        • Zheng Y
        • Carter NP
        • Price CM
        • et al.
        Prenatal diagnosis from maternal blood: simultaneous immunophenotyping and FISH of fetal nucleated erythrocytes isolated by negative magnetic cell sorting.
        J Med Genet. 1993; 30: 1051-1056
        • Gänshirt-Ahlert D
        • Börjesson-Stoll R
        • Burschyk M
        • et al.
        Detection of fetal trisomies 21 and 18 from maternal blood using triple gradient and magnetic cell sorting.
        Am J Reprod Immunol. 1993; 30: 194-201
        • Büsch J
        • Huber P
        • Pflüger E
        • Miltenyi S
        • Holtz J
        • Radbruch A
        Enrichment of fetal cells from maternal blood by high gradient magnetic cell sorting (double MACS) for PCR-based genetic analysis.
        Prenat Diagn. 1994; 14: 1129-1140
        • Cui K
        • Warnes GM
        • Jeffrey R
        • Matthews CD.
        Sex determination of preimplantation embryos by human testis-determining-gene amplification.
        Lancet. 1994; 343: 79-82
        • Douglas GC
        • King BF.
        Isolation of pure villous cytotrophoblast from term human placenta using immunomagnetic microspheres.
        J Immunol Methods. 1989; 119: 259-268
        • Lo Y-MD
        • Patel P
        • Baigent CN
        • et al.
        Prenatal sex determination from maternal peripheral blood using the polymerase chain reaction.
        Hum Genet. 1993; 90: 483-488
        • Adkison LR
        • Andrews RH
        • Vowell NL
        • Koontz WL.
        Improved detection of fetal cells from maternal blood with polymerase chain reaction.
        AM J OBSTET GYNECOL. 1994; 170: 952-955
        • Hamada H
        • Arinami T
        • Kubo T
        • Hamaguchi H
        • Iwasaki H.
        Fetal nucleated cells in maternal peripheral blood: frequency and relationship to gestational age.
        Hum Genet. 1993; 91: 427-432
        • Suzumori K
        • Adachi R
        • Okada S
        • Narukawa T
        • Yagami Y
        • Sonta S.
        Fetal cells in the maternal circulation: detection of Y sequence by gene amplification.
        Obstet Gynecol. 1992; 80: 150-154
        • Nakagome Y
        • Nagafuchi S
        • Nakahori Y.
        Prenatal sex determination.
        Lancet. 1990; 335: 291
        • Goodfellow PN
        • Lovell-Badge R.
        SRY and sex determination in mammals.
        Annu Rev Genet. 1993; 27: 71-92
        • Bianchi DW
        • Shuber AP
        • DeMaria MA
        • Fougner AC
        • Klinger KW.
        Fetal cells in maternal blood: determination of purity and yield by quantitative polymerase chain reaction.
        AM J OBSTET GYNECOL. 1994; 171: 922-926
        • van Wijk IJ
        • van Vugt JMG
        • Könst AAM
        • Mulders MAM
        • Nieuwint AWM
        • Oudejans CBM
        Multiparameter in situ analysis of trophoblast cells in mixed cell populations by combined DNA and RNA in situ hybridization.
        J Histochem Cytochem. 1995; 43: 709-714