Advertisement

Fetal RhD genotyping in fetal cells flow sorted from maternal blood

      Abstract

      OBJECTIVE: The aim of this study was to determine the accuracy of noninvasive fetal RhD genotyping by fetal cell isolation from maternal blood. STUDY DESIGN: Candidate fetal cells from 18 pregnant women (one twin gestation) were flow-sorted. Polymerase chain reaction amplification of a 261 bp fragment of the RhD gene was performed on sorted fetal cells. The presence of amplified product was considered predictive of the RhD-positive genotype in the fetus. RESULTS: Sixteen of the 19 fetal RhD genotypes were correctly predicted in fetal cells isolated from maternal blood (10 were Rh positive, 6 were Rh negative). In 3 cases no amplification products were detected in RhD-positive fetuses. The association between presence of the fragment and RhD-positive genotype was significant (p = 0.003, Fisher's exact test). CONCLUSIONS: Noninvasive prenatal diagnosis of the fetal RhD genotype is feasible. Absence of amplification products in the reaction requires confirmation that fetal material is present. Improvements in fetal cell purity and yield should increase diagnostic accuracy, although the current protocol has a positive predictive value of 100% and a negative predictive value of 67%. (AM J OBSTET GYNECOL 1996;174:818-22.)

      Keywords

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to American Journal of Obstetrics & Gynecology
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Chavez GF
        • Mulinare J
        • Edmonds LD
        Epidemiology of Rh hemolytic disease of the newborn in the United States.
        JAMA. 1991; 265: 3270-3274
        • Agre P
        • Cartron JP
        Molecular biology of the Rh antigens.
        Blood. 1991; 78: 551-563
        • Chérif-Zahar B
        • Mattei MG
        • Le Van Kim C
        • Bailly P
        • Cartron JP
        • Colin Y
        Localization of the human Rh blood group gene structure to chromosome region 1p34.3-1p36.1 by in situ hybridization.
        Hum Genet. 1991; 86: 398-400
        • Le Van Kim C
        • Mouro I
        • Raynal V
        • et al.
        Molecular cloning and primary structure of the human blood group Rh D polypeptide.
        Proc Natl Acad Sci U S A. 1992; 89: 10925-10929
        • Colin Y
        • Chérif-Zahar B
        • Le Van Kim C
        • Raynal V
        • Van Huffel V
        • Cartron JP
        Genetic basis of the Rh D-positive and Rh D-negative blood group polymorphisms as determined by Southern analysis.
        Blood. 1991; 78: 2747-2752
        • Bennett PR
        • Le Van Kim C
        • Colin Y
        • et al.
        Prenatal determination of fetal RhD type by DNA amplification.
        N Engl J Med. 1993; 329: 607-610
        • Rossiter JP
        • Blakemore KJ
        • Kickler TS
        • et al.
        The use of polymerase chain reaction to determine fetal RhD status.
        AM J OBSTET GYNECOL. 1994; 171: 1047-1051
        • Brambati B
        • Anelli MC
        • Tului L
        • Colombo G.
        Fetal RhD typing by DNA amplification in chorionic villus sample.
        Lancet. 1994; 344: 959-960
        • Van den Veyver IB
        • Chong SS
        • Cota J
        • et al.
        Single cell analysis of the RhD blood type for use in preimplantation diagnosis in the prevention of severe hemolytic disease of the newborn.
        AM J OBSTET GYNECOL. 1995; 172: 533-540
        • Lo Y-MD
        • Bowell PJ
        • Selinger M
        • et al.
        Prenatal determination of fetal Rh D status by analysis of peripheral blood of rhesus negative mothers.
        Lancet. 1993; 341: 1147-1148
        • Lo Y-MD
        • Bowell PJ
        • Selinger M
        • et al.
        Prenatal determination of fetal Rhesus D status by DNA amplification of peripheral blood of Rhesus-negative mothers.
        Ann N Y Acad Sci. 1994; 731: 229-236
        • Bianchi DW
        • Zickwolf GK
        • Yih MC
        • et al.
        Erythroid-specific antibodies enchance detection of fetal nucleated erythrocytes in maternal blood.
        Prenat Diagn. 1993; 13: 293-300
        • Parks DR
        • Herzenberg LA
        Fetal cells from maternal blood: their selection and prospects for use in prenatal diagnosis.
        Methods Cell Biol. 1986; 26: 277-295
        • 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
        • Walsh SP
        • Metzger DA
        • Higuchi R
        Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material.
        Biotechniques. 1991; 10: 506-513
        • Simsek S
        • Bleeker PMM
        • von dem Borne AEGK
        Prenatal determination of fetal RhD type.
        N Engl J Med. 1994; 330: 795
        • Geifman-Holtzman O
        • Holtzman EJ
        • Vadnais TJ
        • Phillips VE
        • Capeless EL
        • Bianchi DW
        Detection of fetal HLA DQ alpha sequences in maternal blood: a gender-independent technique of fetal cell identification.
        Prenat Diagn. 1995; 15: 261-268