Prenatal Diagnosis of Fetal RhD Status by Molecular Analysis of Maternal Plasma
Y.M. Dennis Lo, M.R.C.P., N. Magnus Hjelm, F.R.C.Path., Carrie Fidler, Ph.D., Ian L. Sargent, Ph.D., Michael F. Murphy, F.R.C.Path., Paul F. Chamberlain, M.D., Priscilla M.K. Poon, Ph.D., Christopher W.G. Redman, F.R.C.P., and James S. Wainscoat, F.R.C.Path.
Background The ability to determine fetal RhD status noninvasivelyis useful in the treatment of RhD-sensitized pregnant womenwhose partners are heterozygous for the RhD gene. The recentdemonstration of fetal DNA in maternal plasma raises the possibilitythat fetal RhD genotyping may be possible with the use of maternalplasma.
Methods We studied 57 RhD-negative pregnant women and theirsingleton fetuses. DNA extracted from maternal plasma was analyzedfor the RhD gene with a fluorescence-based polymerase-chain-reaction(PCR) test sensitive enough to detect the RhD gene in a singlecell. Fetal RhD status was determined directly by serologicanalysis of cord blood or PCR analysis of amniotic fluid.
Results Among the 57 RhD-negative women, 12 were in their firsttrimester of pregnancy, 30 were in their second trimester, and15 were in their third trimester. Thirty-nine fetuses were RhD-positive,and 18 were RhD-negative. In the samples obtained from womenin their second or third trimester of pregnancy, the resultsof RhD PCR analysis of maternal plasma DNA were completely concordantwith the results of serologic analysis. Among the maternal plasmasamples collected in the first trimester, 2 contained no RhDDNA, but the fetuses were RhD-positive; the results in the other10 samples were concordant (7 were RhD-positive, and 3 RhD-negative).
Conclusions Noninvasive fetal RhD genotyping can be performedrapidly and reliably with the use of maternal plasma beginningin the second trimester of pregnancy.
Source Information
From the Department of Chemical Pathology, Chinese University of Hong Kong, Prince of Wales Hospital, Hong Kong, China (Y.M.D.L., N.M.H., P.M.K.P.); and the Department of Hematology (C.F., M.F.M., J.S.W.) and the Nuffield Department of Obstetrics and Gynecology (I.L.S., P.F.C., C.W.G.R.), John Radcliffe Hospital, Oxford, United Kingdom.
Address reprint requests to Dr. Lo at the Department of Chemical Pathology, Rm. 38023, Clinical Sciences Bldg., Prince of Wales Hospital, 30–32 Ngan Shing St., Hong Kong, China.
Papageorgiou, E. A., Fiegler, H., Rakyan, V., Beck, S., Hulten, M., Lamnissou, K., Carter, N. P., Patsalis, P. C.
(2009). Sites of Differential DNA Methylation between Placenta and Peripheral Blood: Molecular Markers for Noninvasive Prenatal Diagnosis of Aneuploidies. Am. J. Pathol.
174: 1609-1618
[Abstract][Full Text]
Wright, C. F., Burton, H.
(2009). The use of cell-free fetal nucleic acids in maternal blood for non-invasive prenatal diagnosis. Hum Reprod Update
15: 139-151
[Abstract][Full Text]
Lun, F. M. F., Chiu, R. W. K., Allen Chan, K. C., Yeung Leung, T., Kin Lau, T., Dennis Lo, Y. M.
(2008). Microfluidics Digital PCR Reveals a Higher than Expected Fraction of Fetal DNA in Maternal Plasma. Clin. Chem.
54: 1664-1672
[Abstract][Full Text]
Dennis Lo, Y. M., Chiu, R. W. K.
(2008). Noninvasive Prenatal Diagnosis of Fetal Chromosomal Aneuploidies by Maternal Plasma Nucleic Acid Analysis. Clin. Chem.
54: 461-466
[Abstract][Full Text]
Chim, S. S.C., Shing, T. K.F., Hung, E. C.W., Leung, T.-y., Lau, T.-k., Chiu, R. W.K., Dennis Lo, Y.M.
(2008). Detection and Characterization of Placental MicroRNAs in Maternal Plasma. Clin. Chem.
54: 482-490
[Abstract][Full Text]
Tong, Y. K., Chiu, R. W.K., Leung, T. Y., Ding, C., Lau, T. K., Leung, T. N., Lo, Y.M. D.
(2007). Detection of Restriction Enzyme Digested Target DNA by PCR Amplification Using a Stem-Loop Primer: Application to the Detection of Hypomethylated Fetal DNA in Maternal Plasma. Clin. Chem.
53: 1906-1914
[Abstract][Full Text]
Lo, Y. M. D., Lun, F. M. F., Chan, K. C. A., Tsui, N. B. Y., Chong, K. C., Lau, T. K., Leung, T. Y., Zee, B. C. Y., Cantor, C. R., Chiu, R. W. K.
(2007). From the Cover: Digital PCR for the molecular detection of fetal chromosomal aneuploidy. Proc. Natl. Acad. Sci. USA
104: 13116-13121
[Abstract][Full Text]
Tong, Y. K., Ding, C., Chiu, R. W.K., Gerovassili, A., Chim, S. S.C., Leung, T. Y., Leung, T. N., Lau, T. K., Nicolaides, K. H., Lo, Y.M. D.
(2006). Noninvasive Prenatal Detection of Fetal Trisomy 18 by Epigenetic Allelic Ratio Analysis in Maternal Plasma: Theoretical and Empirical Considerations. Clin. Chem.
52: 2194-2202
[Abstract][Full Text]
Chan, K.C. A., Ding, C., Gerovassili, A., Yeung, S. W., Chiu, R. W.K., Leung, T. N., Lau, T. K., Chim, S. S.C., Chung, G. T.Y., Nicolaides, K. H., Lo, Y.M. D.
(2006). Hypermethylated RASSF1A in Maternal Plasma: A Universal Fetal DNA Marker that Improves the Reliability of Noninvasive Prenatal Diagnosis. Clin. Chem.
52: 2211-2218
[Abstract][Full Text]
Okazaki, S, Sekizawa, A, Purwosunu, Y, Iwasaki, M, Farina, A, Okai, T
(2006). Measurement of mRNA of trophoblast-specific genes in cellular and plasma components of maternal blood.. J. Med. Genet.
43: e47-e47
[Abstract][Full Text]
Oepkes, D., Seaward, P. G., Vandenbussche, F. P.H.A., Windrim, R., Kingdom, J., Beyene, J., Kanhai, H. H.H., Ohlsson, A., Ryan, G., the DIAMOND Study Group,
(2006). Doppler ultrasonography versus amniocentesis to predict fetal anemia.. NEJM
355: 156-164
[Abstract][Full Text]
Chan, K.C. A., Hui, A. B.Y., Wong, N., Lau, T. K., Leung, T. N., Lo, K.-W., Lo, Y.M. D.
(2005). Investigation of the Genomic Representation of Plasma DNA in Pregnant Women by Comparative Genomic Hybridization Analysis: A Feasibility Study. Clin. Chem.
51: 2398-2401
[Full Text]
Chim, S. S. C., Tong, Y. K., Chiu, R. W. K., Lau, T. K., Leung, T. N., Chan, L. Y. S., Oudejans, C. B. M., Ding, C., Lo, Y. M. D.
(2005). Detection of the placental epigenetic signature of the maspin gene in maternal plasma. Proc. Natl. Acad. Sci. USA
102: 14753-14758
[Abstract][Full Text]
Zimmermann, B., El-Sheikhah, A., Nicolaides, K., Holzgreve, W., Hahn, S.
(2005). Optimized Real-Time Quantitative PCR Measurement of Male Fetal DNA in Maternal Plasma. Clin. Chem.
51: 1598-1604
[Abstract][Full Text]
Chung, G. T.Y., Chiu, R. W.K., Chan, K.C. A., Lau, T. K., Leung, T. N., Chan, L. W., Lo, Y.M. D.
(2005). Detrimental Effect of Formaldehyde on Plasma RNA Detection. Clin. Chem.
51: 1074-1076
[Full Text]
Lo, Y.M. D.
(2005). Recent Advances in Fetal Nucleic Acids in Maternal Plasma. J. Histochem. Cytochem.
53: 293-296
[Abstract][Full Text]
Hromadnikova, I., Vechetova, L., Vesela, K., Benesova, B., Doucha, J., Vlk, R.
(2005). Non-invasive Fetal RHD and RHCE Genotyping Using Real-time PCR Testing of Maternal Plasma in RhD-negative Pregnancies. J. Histochem. Cytochem.
53: 301-305
[Abstract][Full Text]
Gonzalez-Gonzalez, C., Garcia-Hoyos, M., Trujillo-Tiebas, M. J., Lorda-Sanchez, I., de Alba, M. R., Infantes, F., Gallego, J., Diaz-Recasens, J., Ayuso, C., Ramos, C.
(2005). Application of Fetal DNA Detection in Maternal Plasma: A Prenatal Diagnosis Unit Experience. J. Histochem. Cytochem.
53: 307-314
[Abstract][Full Text]
Chung, G. T.Y., Chiu, R. W.K., Chan, K.C. A., Lau, T. K., Leung, T. N., Lo, Y.M. D.
(2005). Lack of Dramatic Enrichment of Fetal DNA in Maternal Plasma by Formaldehyde Treatment. Clin. Chem.
51: 655-658
[Full Text]
Gingeras, T. R., Higuchi, R., Kricka, L. J., Lo, Y.M. D., Wittwer, C. T.
(2005). Fifty Years of Molecular (DNA/RNA) Diagnostics. Clin. Chem.
51: 661-671
[Full Text]
Li, Y., Di Naro, E., Vitucci, A., Zimmermann, B., Holzgreve, W., Hahn, S.
(2005). Detection of Paternally Inherited Fetal Point Mutations for {beta}-Thalassemia Using Size-Fractionated Cell-Free DNA in Maternal Plasma. JAMA
293: 843-849
[Abstract][Full Text]
Boddy, J. L., Gal, S., Malone, P. R., Harris, A. L., Wainscoat, J. S.
(2005). Prospective Study of Quantitation of Plasma DNA Levels in the Diagnosis of Malignant versus Benign Prostate Disease. Clin. Cancer Res.
11: 1394-1399
[Abstract][Full Text]
Bischoff, F. Z., Lewis, D. E., Simpson, J. L.
(2005). Cell-free fetal DNA in maternal blood: kinetics, source and structure. Hum Reprod Update
11: 59-67
[Abstract][Full Text]
Tsui, N B Y, Chim, S S C, Chiu, R W K, Lau, T K, Ng, E K O, Leung, T N, Tong, Y K, Chan, K C A, Lo, Y M D
(2004). Systematic micro-array based identification of placental mRNA in maternal plasma: towards non-invasive prenatal gene expression profiling. J. Med. Genet.
41: 461-467
[Full Text]
Masuzaki, H, Miura, K, Yoshiura, K-i, Yoshimura, S, Niikawa, N, Ishimaru, T
(2004). Detection of cell free placental DNA in maternal plasma: direct evidence from three cases of confined placental mosaicism. J. Med. Genet.
41: 289-292
[Full Text]
Simpson, J. L., Bischoff, F.
(2004). Cell-Free Fetal DNA in Maternal Blood: Evolving Clinical Applications. JAMA
291: 1135-1137
[Full Text]
Rijnders, R. J.P., Christiaens, G. C.M.L., Soussan, A. A., van der Schoot, C. E., Invernizzi, P., Simoni, G., Podda, M.
(2004). Cell-Free Fetal DNA Is Not Present in Plasma of Nonpregnant Mothers * Authors of the article cited above respond:. Clin. Chem.
50: 679-681
[Full Text]
Chan, K.C. A., Zhang, J., Hui, A. B.Y., Wong, N., Lau, T. K., Leung, T. N., Lo, K.-W., Huang, D. W.S., Lo, Y.M. D.
(2004). Size Distributions of Maternal and Fetal DNA in Maternal Plasma. Clin. Chem.
50: 88-92
[Abstract][Full Text]
Lam, N. Y.L., Rainer, T. H., Chiu, R. W.K., Joynt, G. M., Lo, Y.M. D.
(2004). Plasma Mitochondrial DNA Concentrations after Trauma. Clin. Chem.
50: 213-216
[Full Text]
Ng, E. K.O., Leung, T. N., Tsui, N. B.Y., Lau, T. K., Panesar, N. S., Chiu, R. W.K., Lo, Y.M. D.
(2003). The Concentration of Circulating Corticotropin-releasing Hormone mRNA in Maternal Plasma Is Increased in Preeclampsia. Clin. Chem.
49: 727-731
[Abstract][Full Text]
Chan, M. H.M., Chow, K. M., Chan, A. T.C., Leung, C. B., Chan, L. Y.S., Chow, K. C.K., Lam, C. W., Lo, Y.M. D.
(2003). Quantitative Analysis of Pleural Fluid Cell-free DNA as a Tool for the Classification of Pleural Effusions. Clin. Chem.
49: 740-745
[Abstract][Full Text]
Ng, E. K. O., Tsui, N. B. Y., Lau, T. K., Leung, T. N., Chiu, R. W. K., Panesar, N. S., Lit, L. C. W., Chan, K.-W., Lo, Y. M. D.
(2003). From the Cover: mRNA of placental origin is readily detectable in maternal plasma. Proc. Natl. Acad. Sci. USA
100: 4748-4753
[Abstract][Full Text]
Rainer, T. H., Wong, L. K.S., Lam, W., Yuen, E., Lam, N. Y.L., Metreweli, C., Lo, Y.M. D.
(2003). Prognostic Use of Circulating Plasma Nucleic Acid Concentrations in Patients with Acute Stroke. Clin. Chem.
49: 562-569
[Abstract][Full Text]
Chan, L. Y.S., Leung, T. N., Chan, K.C. A., Tai, H.-L., Lau, T. K., Wong, E. M.C., Lo, Y.M. D.
(2003). Serial Analysis of Fetal DNA Concentrations in Maternal Plasma in Late Pregnancy. Clin. Chem.
49: 678-680
[Full Text]
Chui, D. H. K., Fucharoen, S., Chan, V.
(2003). Hemoglobin H disease: not necessarily a benign disorder. Blood
101: 791-800
[Full Text]
Costa, J.-M., Benachi, A., Gautier, E.
(2002). New Strategy for Prenatal Diagnosis of X-Linked Disorders. NEJM
346: 1502-1502
[Full Text]
Bianchi, D. W.
(2002). Prenatal Exclusion of Recessively Inherited Disorders: Should Maternal Plasma Analysis Precede Invasive Techniques?. Clin. Chem.
48: 689-690
[Full Text]
Chiu, R. W.K., Lau, T. K., Cheung, P. T., Gong, Z. Q., Leung, T. N., Lo, Y.M. D.
(2002). Noninvasive Prenatal Exclusion of Congenital Adrenal Hyperplasia by Maternal Plasma Analysis: A Feasibility Study. Clin. Chem.
48: 778-780
[Full Text]
Costa, J.-M., Ernault, P.
(2002). Automated Assay for Fetal DNA Analysis in Maternal Serum. Clin. Chem.
48: 679-680
[Full Text]
Poon, L. L.M., Leung, T. N., Lau, T. K., Chow, K. C.K., Lo, Y.M. D.
(2002). Differential DNA Methylation between Fetus and Mother as a Strategy for Detecting Fetal DNA in Maternal Plasma. Clin. Chem.
48: 35-41
[Abstract][Full Text]
Sekizawa, A., Kondo, T., Iwasaki, M., Watanabe, A., Jimbo, M., Saito, H., Okai, T.
(2001). Accuracy of Fetal Gender Determination by Analysis of DNA in Maternal Plasma. Clin. Chem.
47: 1856-1858
[Full Text]
Chiu, R. W. K., Poon, L. L. M., Lau, T. K., Leung, T. N., Wong, E. M. C., Lo, Y. M. D.
(2001). Effects of Blood-Processing Protocols on Fetal and Total DNA Quantification in Maternal Plasma. Clin. Chem.
47: 1607-1613
[Abstract][Full Text]
Kruskall, M. S., Lee, T.-H., Assmann, S. F., Laycock, M., Kalish, L. A., Lederman, M. M., Busch, M. P.
(2001). Survival of transfused donor white blood cells in HIV-infected recipients. Blood
98: 272-279
[Abstract][Full Text]
Chiu, R. W.K., Murphy, M. F., Fidler, C., Zee, B. C.Y., Wainscoat, J. S., Lo, Y.M. D.
(2001). Determination of RhD Zygosity: Comparison of a Double Amplification Refractory Mutation System Approach and a Multiplex Real-Time Quantitative PCR Approach. Clin. Chem.
47: 667-672
[Abstract][Full Text]
Honda, H., Miharu, N., Ohashi, Y., Ohama, K.
(2001). Successful Diagnosis of Fetal Gender Using Conventional PCR Analysis of Maternal Serum. Clin. Chem.
47: 41-46
[Abstract][Full Text]
Leung, T. N., Zhang, J., Lau, T. K., Chan, L. Y.S., Lo, Y.M. D.
(2001). Increased Maternal Plasma Fetal DNA Concentrations in Women Who Eventually Develop Preeclampsia.. Clin. Chem.
47: 137-139
[Full Text]
Lo, Y.M. D.
(2000). Fetal DNA in Maternal Plasma: Biology and Diagnostic Applications. Clin. Chem.
46: 1903-1906
[Abstract][Full Text]
Poon, L. L.M., Leung, T. N., Lau, T. K., Lo, Y.M. D.
(2000). Presence of Fetal RNA in Maternal Plasma. Clin. Chem.
46: 1832-1834
[Full Text]
Lo, Y.M. D.
(2000). Molecular Testing of Urine: Catching DNA on the Way Out. Clin. Chem.
46: 1039-1040
[Full Text]
Botezatu, I., Serdyuk, O.'g., Potapova, G., Shelepov, V., Alechina, R., Molyaka, Y., Anan'ev, V., Bazin, I., Garin, A., Narimanov, M., Knysh, V., Melkonyan, H., Umansky, S., Lichtenstein, A.
(2000). Genetic Analysis of DNA Excreted in Urine: A New Approach for Detecting Specific Genomic DNA Sequences from Cells Dying in an Organism. Clin. Chem.
46: 1078-1084
[Abstract][Full Text]
van Wijk, I. J., de Hoon, A. C., Jurhawan, R., Tjoa, M. L., Griffioen, S., Mulders, M. A.M., van Vugt, J. M.G., Oudejans, C. B.M.
(2000). Detection of Apoptotic Fetal Cells in Plasma of Pregnant Women. Clin. Chem.
46: 729-731
[Full Text]
Avent, N. D., Reid, M. E.
(2000). The Rh blood group system: a review. Blood
95: 375-387
[Abstract][Full Text]
Saade, G. R.
(2000). Noninvasive Testing for Fetal Anemia. NEJM
342: 52-53
[Full Text]
Singleton, B. K., Green, C. A., Avent, N. D., Martin, P. G., Smart, E., Daka, A., Narter-Olaga, E. G., Hawthorne, L. M., Daniels, G.
(2000). The presence of an RHD pseudogene containing a 37 base pair duplication and a nonsense mutation in Africans with the Rh D-negative blood group phenotype. Blood
95: 12-18
[Abstract][Full Text]
Tang, N. L.S., Leung, T. N., Zhang, J., Lau, T. K., Lo, Y.M. D.
(1999). Detection of Fetal-derived Paternally Inherited X-Chromosome Polymorphisms in Maternal Plasma. Clin. Chem.
45: 2033-2035
[Full Text]
Lo, Y.M. D., Lau, T. K., Zhang, J., Leung, T. N., Chang, A. M.Z., Hjelm, N. M., Elmes, R. S., Bianchi, D. W.
(1999). Increased Fetal DNA Concentrations in the Plasma of Pregnant Women Carrying Fetuses with Trisomy 21. Clin. Chem.
45: 1747-1751
[Abstract][Full Text]
Silva, J. M., Dominguez, G., Garcia, J. M., Gonzalez, R., Villanueva, M. J., Navarro, F., Provencio, M., San Martin, S., Espana, P., Bonilla, F.
(1999). Presence of Tumor DNA in Plasma of Breast Cancer Patients: Clinicopathological Correlations. Cancer Res.
59: 3251-3256
[Abstract][Full Text]
Bowman, J. M.
(1998). RhD Hemolytic Disease of the Newborn. NEJM
339: 1775-1777
[Full Text]
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