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Amplicon Sequencing-Based Noninvasive Fetal Genotyping for RHD-Positive D Antigen-Negative Alleles

Published on Oct 1, 2019in Clinical Chemistry6.891
· DOI :10.1373/clinchem.2019.307074
Ken Takahashi1
Estimated H-index: 1
(Jikei University School of Medicine),
Ohsuke Migita8
Estimated H-index: 8
(St. Marianna University School of Medicine)
+ 9 AuthorsKenichiro Hata32
Estimated H-index: 32
Abstract
Background: To avoid hemolytic disease of the fetus and newborn resulting from maternal alloantibodies against fetal Rh antigens, anti-D immunoglobulin is routinely administered to RhD-negative pregnant women in Japan. Fetal RHD genotyping using cell-free DNA may prevent unnecessary antibody administration; however, current PCR-based methods, which detect RHD deletion, do not address the higher rates of RHD-positive D antigen-negative alleles in nonwhite populations without additional inspections. Methods: We developed an amplicon-sequencing method that could estimate the type of paternally inherited fetal RHD allele from 4 major RHD alleles in the Japanese population: the D antigen-positive allele (RHD*01, 92.9%) and 3 D antigen-negative alleles (RHD*01N.01, 6.6%; RHD*01EL.01, 0.3%; RHD* 01N.04, 0.1%) using cell-free DNA obtained from the blood plasma of pregnant women. Results: The method correctly determined the fetal RhD type even when RhD-negative pregnant women possessed an RHD-positive D antigen-negative allele: RHD*01EL.01 or RHD*01N.04. Conclusions: This method is a reliable noninvasive fetal RHD genotyping method for Japanese and other East Asian populations. The genotyping principle of amplifying 2 different regions using the same primer pair and distinguishing them by their sequence difference during the subsequent mapping procedure is also theoretically applicable to RHD-positive D antigen-negative alleles prevalent in Africans. Therefore, this method offers an opportunity to consider targeted administration of anti-D immunoglobulin to RhD-negative pregnant women in East Asian and African countries and to increase the specificity of the fetal RHD genotyping implemented nationwide in several European countries.
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References40
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#1Yuqin Luo (ZJU: Zhejiang University)H-Index: 1
#2Bei Jia (Southern Medical University)H-Index: 1
Last. Min-Yue Dong (ZJU: Zhejiang University)H-Index: 12
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#1Huiqin Yang (University of Exeter)H-Index: 13
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Background High-throughput non-invasive prenatal testing (NIPT) for fetal Rhesus D (RhD) status could avoid unnecessary treatment with anti-D immunoglobulin for RhD-negative women found to be carrying an RhD-negative fetus. We aimed to assess the diagnostic accuracy of high-throughput NIPT for fetal RhD status in RhD-negative women not known to be sensitized to the RhD antigen, by performing a systematic review and meta-analysis.
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#2S. Shahrukh Hashmi (University of Texas Health Science Center at Houston)H-Index: 13
Last. Michael Bebbington (University of Texas Health Science Center at San Antonio)H-Index: 46
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The determination of foetal Rhesus D (RHD) status allows appropriate use of IgRh prophylaxis by restricting its use to cases of RHD feto-maternal incompatibilities. There is a degree of uncertainty about the cost-effectiveness of foetal RHD determination, yet screening programs are being introduced into clinical practice in many countries. This paper evaluates the impact of non-invasive foetal Rhesus D (RHD) status determination on the costs of managing RHD-negative pregnant women and on the app...
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#1Silvia ManfroiH-Index: 4
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Background Foetal RHD genotyping can be predicted by real-time polymerase chain reaction (qPCR) using cell-free foetal DNA extracted from maternal plasma. The object of this study was to determine the diagnostic accuracy and feasibility of non-invasive RHD foetal genotyping, using a commercial multiple-exon assay, as a guide to appropriate administration of targeted antenatal immunoprophylaxis.
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Purpose of reviewIn this review, we analyzed the current literature on noninvasive fetal RHD typing to answer the question whether the administration of RhIg to prevent D-alloimmunization during pregnancy can be safely guided by fetal RHD typing.Recent findingsRecently the first centers that impleme
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#1Fiona L. Mackie (University of Birmingham)H-Index: 5
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Background Cell-free fetal DNA (cffDNA) non-invasive prenatal testing (NIPT) is rapidly expanding, and is being introduced at varying rates depending on country and condition. Objectives Determine accuracy of cffDNA-based NIPT for all conditions. Evaluate influence of other factors on test performance. Search strategy Medline, Embase, CINAHL, Cochrane Library, from 1997 to April 2015. Selection criteria Cohort studies reporting cffDNA-based NIPT performance in singleton pregnancies. Data collect...
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Objective To determine the accuracy of non-invasive fetal testing for the RHD gene in week 27 of pregnancy as part of an antenatal screening programme to restrict anti-D immunoglobulin use to women carrying a child positive for RHD. Design Prospectively monitoring of fetal RHD testing accuracy compared with serological cord blood typing on introduction of the test. Fetal RHD testing was performed with a duplex real time quantitative polymerase chain reaction, with cell-free fetal DNA isolated fr...
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Last. C. Ellen van der Schoot (UvA: University of Amsterdam)H-Index: 64
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To guide anti-D prophylaxis, Dutch D- pregnant women are offered a quantitative fetal-RHD-genotyping assay to determine the RHD status of their fetus. This allowed us to determine the frequency of different maternal RHD variants in 37 782 serologically D- pregnant women. A variant allele is present in at least 0.96% of Dutch D- pregnant women The D- serology could be confirmed after further serological testing in only 54% of these women, which emphasizes the potential relevance of genotyping of ...
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