Prenatal determination of fetal RhD status by analysis of peripheral blood of rhesus negative mothers

Lo, Y.M. Dennis; Bowell, P. J.; Selinger, M.; Mackenzie, I. Z.; Chamberlain, P.; Gillmer, M. D. G.; Littlewood, Timothy; Fleming, K. A.; Wainscoat, James S.

doi:10.1016/0140-6736(93)93161-s

Cited by 139 publications

(49 citation statements)

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“…F and R indicate forward and reverse primer, respectively Determined by the DNA-based sequence-specific primers polymerase chain reaction typing method described in the text almost invariably, lack the RHD gene (Daniels 1995). This finding not only explains finally why no postulated Rh d antigen has ever been shown, but also makes it possible to determine the RhD status of fetuses at risk for HDN, using a DNA-based SSP-PCR technique (Lo et al 1993;Denomme et al 1999). Later studies, however, have revealed that RhD-negative individuals may have different genetic backgrounds.…”

Section: Discussionmentioning

confidence: 89%

Genetic polymorphism of RhD-negative associated haplotypes in the Chinese

Lei

Chen

et al. 2000

J Hum Genet

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The Rh blood group is the most polymorphic human blood group system, and is clinically significant in transfusion medicine. Individuals are classified as Rhpositive and Rh-negative depending on the presence or absence of the D antigen on the red cell surface. The RhDnegative trait could be generated by multiple genetic mechanisms, which have been shown to be ethnic groupdependent. In this study, we evaluated the status of seven RHD-specific exons (exons 3, 4, 5, 6, 7, 9, and 10) and RH intron 4 in 119 Chinese blood donors, using the sequencespecific primers polymerase chain reaction (SSP-PCR). Of the 87 individuals who were RhD-negative, 52 with the ce/ ce, ce/cE, or Ce/ce genotype (60%) lacked the above seven RHD exons; 22 with the Ce/Ce or Ce/ce genotype (25%) had all the RHD exons examined; 13 with the Ce/ce genotype (15%) carried at least one RHD exon. Antigen association analysis suggested the existence of a novel class of RhD-negative associated haplotypes in the Chinese, tentatively denoted D(nf)Ce. The D(nf)Ce haplotype consisted of a normal RHCe allele and a nonfunctional RHD gene, which vary depending on the structure of the RHD gene. Among the RhD-negative Chinese, the estimated frequencies of the dce, dCe, and D(nf)Ce haplotypes were 0.7500, 0.0465, and 0.2035, respectively. No statistically significant deviation from Hardy-Weinberg equilibrium was observed using this genetic model.

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Section: Discussionmentioning

confidence: 89%

Genetic polymorphism of RhD-negative associated haplotypes in the Chinese

Lei

Chen

et al. 2000

J Hum Genet

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“…The primers D4 and D5 used in method I, were derived from the PCR system of Le Van Kim et al (1992) and Bennett et al (1993). The D4 forward primer was derived from primer A3 which has been 5 0 -extended with six additional bases to increase the PCR annealing temperature to 58ЊC, and the reverse primer D5 was chosen 99 bases 3 0 downstrean to the initial RhXIII/A4 primer and corresponded to primers RDA3/RD2 of Lo et al (1993). The D5 primer is located in the 3 0 untranslated region specific of the RhD cDNA to avoid undesirable amplification of RHCE sequences.…”

Section: Methodsmentioning

confidence: 99%

“…However, molecular diagnosis of the Rh status is now possible, since the two homologous RH genes (D and CE) have been cloned (for reviews see Cartron & Agre, 1993;Anstee & Tanner, 1993), and it has been demonstrated that the presence or absence of the D gene in the genome determines the genetic basis of the polymorphism associated with the D-positive and D-negative phenotypes, respectively (Colin et al, 1991). Based on these findings, and because Southern techniques (Colin et al, 1991;Hyland et al, 1994a;Huang et al, 1996) are too time-consuming and complex for routine use when other methods are available, a prenatal determination of the fetal RHD type by DNA amplification has been proposed (Bennett et al, 1993;Lo et al, 1993). However, examination of blood donors (Simsek et al, 1994(Simsek et al, , 1995 and speculations on RH gene rearrangements have raised some doubts about the PCR-based method, using primers to amplify a non-coding region of the RHD gene exon 10.…”

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confidence: 99%

Specificity and sensitivity of RHD genotyping methods by PCR‐based DNA amplification

Jt¹,

Kim

Mouro

et al. 1997

Br J Haematol

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Summary.We have compared the sensitivity and specificity of four PCR methods of RHD gene detection using different sets of primers located in the regions of highest divergence between the RHD and RHCE genes, notably exon 10 (method I), exon 7 (method II), exon 4 (method III) and intron 4 (method IV). Methods I-III were the most sensitive and gave a detectable signal with D-pos/D-neg mixtures containing only 0 . 001% D-positive cells. Moreover, method II could detect the equivalent DNA amount present in only three nucleated cells in the assay without hybridization of PCR products, whereas the sensitivity of the other methods was 10-50 times less. Investigation of D variants indicated that false-negative results were obtained with method II (D IVb variant), method III (D VI and DFR variants) and method IV (D VI variants), but not method I. Weak D (D u ) was correctly detected as D-positive by all methods, but most cases of Rh null appeared as false-positives, as they carry normal RH genes that are not phenotypically expressed. Some false-positive results were obtained with method I in a few Caucasian DNA samples serotyped as RhD-neg but carrying a C-or E-allele, whereas a high incidence of false-positives was found among non-Caucasian Rh-negative samples by all methods. In the Caucasian population, however, we found a full correlation between the predicted genotype and observed phenotype at birth of 92 infants. Although we routinely use the four methods for RHD genotyping, a PCR strategy based on at least two methods is recommended.

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“…3,4 These initial studies were followed by multiple reports of successful prenatal diagnosis of fetal RhD type and other fetal conditions by using fetal DNA obtained from peripheral maternal blood. [5][6][7] Fetal DNA enters the maternal circulation either as free DNA form or as fetal-derived cells.…”

Section: Introductionmentioning

confidence: 99%

Noninvasive fetal RhCE genotyping from maternal blood

Geifman-Holtzman

Grotegut

Gaughan

et al. 2008

BJOG

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Background The successful prevention of RhD disease has brought attention to other red blood cells' antigens causing alloimmunisation including RhC/c and RhE/e. Prenatal diagnosis of fetal Rh genotype from maternal blood is in clinical use in Europe but not in the USA.Objective To estimate the collective reported diagnostic accuracy of fetal RhCE genotyping from peripheral maternal blood and compare the results of genotyping when fetal cells and free fetal DNA (FfDNA) are used.Search strategy English-written literature describing fetal RhCE determination from maternal blood using fetal cells or FfDNA was performed using medical subject headings and text words. The sources included Pubmed (1966Pubmed ( -2007, Ovid , CINAHL, The Cochrane Library, ACP Journal Club and OCLC. Key words were prenatal diagnosis, fetal RhCE, fetal DNA in maternal blood and alloimmunisation.Selection criteria A study was considered eligible if it described fetal RhCE type determination using maternal peripheral blood reported in the English literature. Abstracts were excluded.Data collection and analysis From each study, we determined the number of samples tested, fetal RhCE genotype, the source of the fetal DNA, gestational age, presence of alloimmunisation and confirmation of fetal RhCE type. Exclusions and inclusions were noted. We calculated composite estimates of accuracy using a weighted random effects model. We assessed the papers against an international quality, STARD checklist which is standards for reporting studies of diagnostic accuracy.Main results We identified 20 protocols in six English-written publications reporting fetal RhC/c (seven protocols) and/or E/e (13 protocols) genotyping using DNA obtained from maternal blood for a total of 369 samples. For RhC/c, 176 samples were tested and for RhE/e, 193 samples were tested. Accuracy was determined for each study and for all studies. The combined accuracy of fetal genotype was 96.3% for RhC/c and 98.2% for RhE/e. Only a few samples of the sorted cells were found to be a source for accurate diagnosis, but plasma was consistently the best source of fetal RhCE genotyping in 147/147 (100%) for RhC/c and 168/168 (100%) for RhE/e.Conclusions The combined accuracy of noninvasive fetal RhC/c or RhE/e determination using maternal peripheral blood is 96.3% and 98.2%, respectively. FfDNA in maternal plasma is a better source for genotyping reported to be 100% correct for both RHCE genotypes. Further studies and reports of accuracy from laboratories performing the tests are required before prenatal determination of fetal RhC/c or RhE/e genotypes from maternal blood can safely replace the current methods used in the management of the RhC/c or RhE alloimmunised pregnancies.

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Prenatal determination of fetal RhD status by analysis of peripheral blood of rhesus negative mothers

Cited by 139 publications

References 5 publications

Genetic polymorphism of RhD-negative associated haplotypes in the Chinese

Genetic polymorphism of RhD-negative associated haplotypes in the Chinese

Specificity and sensitivity of RHD genotyping methods by PCR‐based DNA amplification

Noninvasive fetal RhCE genotyping from maternal blood

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