Lessons learned from the implementation of non-invasive fetalRHDscreening

Clausen, Frederik Banch

doi:10.1080/14737159.2018.1461562

Cited by 17 publications

(25 citation statements)

References 108 publications

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“…Blood group genotyping is used in clinical practice [13][14][15][16]. However, ABO genotyping is complicated, and despite successful development of assays for fetal RHD genotyping [17][18][19], or more recently for KEL and other non-RhD antigen targets [20][21][22], less work has been done attempting a noninvasive fetal ABO prediction [23]. Due to the huge genetic variation in the ABO locus [24,25], it is difficult to develop a simple assay that will reach 100% accuracy in different ethnic populations.…”

Section: Introductionmentioning

confidence: 99%

Next Generation Sequencing-Based Fetal ABO Blood Group Prediction by Analysis of Cell-Free DNA from Maternal Plasma

Rieneck

Hother

Clausen

et al. 2020

Transfus Med Hemother

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Introduction: ABO blood group incompatibility between a pregnant woman and her fetus as a cause of morbidity or mortality of the fetus or newborn remains an important, albeit rare, risk. When a pregnant woman has a high level of anti-A or anti-B IgG antibodies, the child may be at risk for hemolytic disease of the fetus and newborn (HDFN). Performing a direct prenatal determination of the fetal ABO blood group can provide valuable clinical information. Objective: Here, we report a next generation sequencing (NGS)based assay for predicting the prenatal ABO blood group. Materials and Methods: A total of 26 plasma samples from 26 pregnant women were tested from gestational weeks 12 to 35. Of these samples, 20 were clinical samples and 6 were test samples. Extracted cell-free DNA was PCR-amplified using 2 primer sets followed by NGS. NGS data were analyzed by 2 different methods, FASTQ analysis and a grep search, to ensure robust results. The fetal ABO prediction was compared with the known serological infant ABO type, which was available for 19 samples. Results: There was concordance for 19 of 19 predictable samples where the phenotype information was available and when the analysis was done by the 2 methods. For immunized pregnant women (n = 20), the risk of HDFN was predicted for 12 fetuses, and no risk was predicted for 7 fetuses; one result of the clinical samples was indeterminable. Cloning and sequencing revealed a novel variant harboring the same single nucleotide variations as ABO*O.01.24 with an additional c.220C>T substitution. An additional indeterminable result was found among the 6 test samples and was caused by maternal heterozygosity. The 2 indeterminable samples demonstrated limitations to the assay due to hybrid ABO genes or maternal heterozygosity. Conclusions: We pioneered an NGS-based fetal ABO prediction assay based on a cell-free DNA analysis from maternal plasma and demonstrated its application in a small number of samples. Based on the calculations of variant frequencies and ABO*O.01/ABO*O.02 heterozygote frequency, we estimate that we can assign a reliable fetal ABO type in approximately 95% of the forthcoming clinical samples of type O pregnant women. Despite the vast genetic variations underlying the ABO blood groups, many variants are rare, and prenatal ABO prediction is possible and adds valuable early information for the prevention of ABO HDFN.

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Section: Introductionmentioning

confidence: 99%

Next Generation Sequencing-Based Fetal ABO Blood Group Prediction by Analysis of Cell-Free DNA from Maternal Plasma

Rieneck

Hother

Clausen

et al. 2020

Transfus Med Hemother

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“…As part of a targeted RhIg prophylaxis program for non-immunized RhD-negative women, knowledge of the fetal RhD type helps restrict prophylaxis to those women only who carry an RhD-positive fetus [12,13]. This restriction avoids superfluous exposure to prophylaxis in women carrying an RhD-negative fetus and reduces the overall use of RhIg, which is a limited resource [14,15].…”

Section: Antenatal Rhd Screeningmentioning

confidence: 99%

Laboratory Monitoring of Mother, Fetus, and Newborn in Hemolytic Disease of Fetus and Newborn

Dziegiel¹,

Krog²,

Hansen³

et al. 2021

Transfus Med Hemother

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Background: Laboratory monitoring of mother, fetus, and newborn in hemolytic disease of fetus and newborn (HDFN) aims to guide clinicians and the immunized women to focus on the most serious problems of alloimmunization and thus minimize the consequences of HDFN in general and of anti-D in particular. Here, we present the current approach of laboratory screening and testing for prevention and monitoring of HDFN at the Copenhagen University Hospital in Denmark. Summary: All pregnant women are typed and screened in the 1st trimester. This serves to identify the RhD-negative pregnant women who at gestational age (GA) of 25 weeks are offered a second screen test and a non-invasive fetal RhD prediction. At GA 29 weeks, and again after delivery, non-immunized RhD-negative women carrying an RhD-positive fetus are offered Rh immunoglobulin. If the 1st trimester screen reveals an alloantibody, antenatal investigation is initiated. This also includes RhD-positive women with alloantibodies. Specificity and titer are determined, the fetal phenotype is predicted by non-invasive genotyping based on cell-free DNA (RhD, K, Rhc, RhC, RhE, ABO), and serial monitoring of titer commences. Based on titers and specificity, monitoring with serial peak systolic velocity measurements in the fetal middle cerebral artery to detect anemia will take place. Intrauterine transfusion is given when fetal anemia is suspected. Monitoring of the newborn by titer and survival of fetal red blood cells by flow cytometry will help predict the length of the recovery of the newborn.

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“…2,5 Fetal RHDscreening is now a routine in many countries, resulting in that the fetal RHD status is known with high accuracy prenatally, with a sensitivity of the analysis above 99,9%. 6 To add anti-D prophylaxis in gestational week 38, to ensure a protective concentration of anti-D at term and post term in RhD negative women at risk of immunisation, seems relevant and logistically possible in most programs. Anti-D prophylaxis in gestational week 38 may replace postpartum prophylaxis, and thus make the strategy cost effective.…”

Section: Main Findingsmentioning

confidence: 99%

“…5 During the last decade fetal RHD -typing combined with targeted RAADP only to those carrying an RHD positive fetus, has been proven effective and is implemented in many countries. 6 The strategies of doses and timing of administration of RAADP vary between countries, one single dose of anti-D; 1000-1500 IU in gestational week 28-30 or two doses of 500-625 IU at 28 and 34 weeks are the most common routines. 7,8,9 A second or third dose anti-D is given postdelivery, usually a dose of 1000-1500 IU.…”

Section: Introductionmentioning

confidence: 99%

“…Today, when fetal RHD typing is widely implemented and theRHD type of the fetus is known with high accuracy 6 an alternative strategy could be to administer two RAADP doses, at gestational week 28 and 38, the latter to cover term and post term of the pregnancy. If the postdelivery anti-D dose could be excluded, this strategy could be cost effective and have the potential of high compliance.…”

Section: Introductionmentioning

confidence: 99%

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Improved protection at term with an alternative strategy of prophylactic anti-D administration in pregnancy - a prospective interventional study

Wikman

rtberg

Jalkesten

et al. 2020

Preprint

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Objective: To analyse the proportion of RhD negative women where routine antenatal anti-D prophylaxis (RAADP) in gestational week (gw) 28 is not detectable at delivery and to investigate if a strategy with RAADP administered in gw 28 and 38 result in efficient protection at term, postterm and postdelivery. Design: A retrospective database analysis and a prospective interventional study. Setting: Antenatal centers in the Stockholm region. Sample: RhD negative women carrying an RHD positive fetus, 4280 cases evaluated retrospectively, 39 cases included prospectively. Methods: In a retrospective analysis, RhD negative women with a negative antibody screen at delivery was determined. In 39 pregnancies, quantification of anti-D was analysed before a second dose anti-D was administered in gw 38, and then weekly up to 43 weeks post gestation. Main outcome measures: The proportion of women with non-detectable anti-D at term. The concentration of anti-D measured weekly at term, postterm and postdelivery. Results: In 20,5% (856/4280, retrospective analysis) and 44% (17/39, prospective study) RAADP in gw 28 was below screening detection level, 10 IU/L at term and in 18% (7/39, prospective study) below 1 IU/L in the quantification assay. Anti-D prophylaxis administered in gw 38 showed stable protective levels of anti-D up to 30 days postdelivery, with concentration at delivery 60+34 IU/L (mean+SD). Conclusion: At least one third of the RhD negative women have non-detectable levels of RAADP given at gw 28, at term and postterm. A second dose of RAADP at gw 38 improves the protection.

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Lessons learned from the implementation of non-invasive fetalRHDscreening

Cited by 17 publications

References 108 publications

Next Generation Sequencing-Based Fetal ABO Blood Group Prediction by Analysis of Cell-Free DNA from Maternal Plasma

Next Generation Sequencing-Based Fetal ABO Blood Group Prediction by Analysis of Cell-Free DNA from Maternal Plasma

Laboratory Monitoring of Mother, Fetus, and Newborn in Hemolytic Disease of Fetus and Newborn

Improved protection at term with an alternative strategy of prophylactic anti-D administration in pregnancy - a prospective interventional study

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