Two Reliable Methodical Approaches for Non-Invasive RHD Genotyping of a Fetus from Maternal Plasma

Böhmová, J; Ľubušký, Marek; Holusková, I; Studničková, M; Kratochvilova, Romana; Krejcirikova, Eva; Durdova, V.; Kratochvílová, Tereza; Dušek, Ladislav; Procházka, Martin; Vodička, Radek

doi:10.3390/diagnostics10080564

Cited by 4 publications

(4 citation statements)

References 61 publications

(68 reference statements)

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“…Most laboratories that provide noninvasive cffDNA RHD genotyping perform a real-time PCR test for the presence or deletion of the RHD gene. Our previous study confirmed that for the Czech population RhD negativity is almost exclusively associated with the deletion of the entire RHD gene [26].…”

Section: Discussionsupporting

confidence: 85%

“…RHD tests using real-time PCR and RHCE and KEL using minisequencing have already been introduced at our workplace [ 26 , 27 , 28 ]. One of the aims of the study was to methodically unify both procedures and, in the case of RHCE and KEL , to simplify and speed up the examination.…”

Section: Discussionmentioning

confidence: 99%

“…All of the subjects enrolled in the study signed an informed consent form approved by the Ethics Committee of the University Hospital Olomouc (approval code: 150/10). The samples that had already been tested by validated TaqMan real-time PCR and minisequencing methods [ 26 , 27 , 28 ] were used to validate and assess the ddPCR methodology. The correlation between fetal genotype and phenotype was further verified in the newborn using an immunoassay.…”

Section: Methodsmentioning

confidence: 99%

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Risk Minimization of Hemolytic Disease of the Fetus and Newborn Using Droplet Digital PCR Method for Accurate Fetal Genotype Assessment of RHD, KEL, and RHCE from Cell-Free Fetal DNA of Maternal Plasma

et al. 2021

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The molecular pathology of hemolytic disease of the fetus and newborn (HDFN) is determined by different RHD, RHCE, and KEL genotypes and by blood group incompatibility between the mother and fetus that is caused by erythrocyte antigen presence/absence on the cell surface. In the Czech Republic, clinically significant antierythrocyte alloantibodies include anti-D, anti-K, anti C/c, and anti-E. Deletion of the RHD gene and then three single nucleotide polymorphisms in the RHCE and KEL genes (rs676785, rs609320, and rs8176058) are the most common. The aim of this study is to develop effective and precise monitoring of fetal genotypes from maternal plasma of these polymorphisms using droplet digital (dd)PCR. Fifty-three plasma DNA samples (from 10 to 18 weeks of gestation) were analyzed (10 RHD, 33 RHCE, and 10 KEL). The ddPCR methodology was validated on the basis of the already elaborated and established method of minisequencing and real-time PCR and with newborn phenotype confirmation. The results of ddPCR were in 100% agreement with minisequencing and real-time PCR and also with newborn phenotype. ddPCR can fully replace the reliable but more time-consuming method of minisequencing and real-time PCR RHD examination. Accurate and rapid noninvasive fetal genotyping minimizes the possibility of HDFN developing.

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Section: Discussionsupporting

confidence: 85%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Risk Minimization of Hemolytic Disease of the Fetus and Newborn Using Droplet Digital PCR Method for Accurate Fetal Genotype Assessment of RHD, KEL, and RHCE from Cell-Free Fetal DNA of Maternal Plasma

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“…As noted previously, maternal anti-RhD IgG antibodies (anti-D) that cross the placenta and destroy fetal red blood cells cause anemia in the infant (RBCs). This is referred to as (HDN) [63].…”

Section: Real-time Pcr-applied Biosystemsmentioning

confidence: 99%

RhD detection in transfusion medicine

Ćutuk,

Hromić- Jahjefendić

2022

Bioengineering Studies

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Transfusion medicine represents one of the emerging medicine branches of the last century. With the discovery of isoagglutinin, the main proof has been made that blood group inheritance is encoded in genes which established the theory of four blood groups. Moreover, the presence of certain antibodies, later known as Rh factors, directly reflects the ability to accept or reject of blood group. Several methods of detection of Rh factor have been used in transfusion medicine which helps in diagnostic procedures. This is of great importance, especially during pregnancies or during blood donation processes. Molecular approaches enable the identification and analysis of blood that is negative for specific antigens against which a patient may have generated antibodies, the detection and analysis of antibodies, and the identification of donors who are negative for common antigens.

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RhD-induced immunogenetic disparity between mother and fetus: medical importance and economic effect of using molecular genetic technology

Коноплянников

Тороповский

Viktorov

et al. 2021

Akušerstvo, ginekologiâ i reprodukciâ

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Introduction. The article discusses the problem of timely diagnosis and prevention of Rh-immunity of pregnant women and hemolytic disease of the fetus, which remains relevant today, despite the existence of proven methods of diagnosis, treatment and prevention.Aim: to assess the medico-economic efficiency of non-invasive prenatal diagnostics of the Rh factor (rhesus D antigen, RhD) of the fetus in the mother's blood – RhD-genotyping of the fetus.Materials and Methods. In a retrospective observational study, the results were analyzed of determining the Rh-factor of the fetus in the blood of 4109 Rh-negative pregnant women observed in 41 medical organizations of the Ulyanovsk region in the period 2018–2020. At a gestational age of ≥ 10 weeks, the RhD state of the fetus was determined by polymerase chain reaction. To assess the medical effectiveness of the test, the sensitivity, specificity, predictive value of positive and negative results, and diagnostic accuracy were determined. The data collected during the study were compared with the information obtained after childbirth. To assess the economic efficiency, the difference between the cost of immunization and the cost of determining the D antigen of the Rhesus system (Rh factor) of the fetus was determined.Results. A positive Rh-factor of the fetus was detected in 67.26 % of cases (n = 2793), negative – in 32.74 % (n = 1316). Diagnostic accuracy of the test system "Test-RhD" was 99.40 %, the sensitivity – 99.84 %, the specificity – 97.51 %, the prognostic value of a positive result – 99.43 %, the predictive value of a negative result – 99.28 % with low rates of false positive and false negative results. It has been shown that this study avoids unnecessary immunization costs for all Rh-negative pregnant women.Conclusion. Analysis of the diagnostic characteristics and cost-effectiveness of the RhD test indicate the high medical significance of the method and make it possible to recommend its wider use.

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Two Reliable Methodical Approaches for Non-Invasive RHD Genotyping of a Fetus from Maternal Plasma

Cited by 4 publications

References 61 publications

Risk Minimization of Hemolytic Disease of the Fetus and Newborn Using Droplet Digital PCR Method for Accurate Fetal Genotype Assessment of RHD, KEL, and RHCE from Cell-Free Fetal DNA of Maternal Plasma

Risk Minimization of Hemolytic Disease of the Fetus and Newborn Using Droplet Digital PCR Method for Accurate Fetal Genotype Assessment of RHD, KEL, and RHCE from Cell-Free Fetal DNA of Maternal Plasma

RhD detection in transfusion medicine

RhD-induced immunogenetic disparity between mother and fetus: medical importance and economic effect of using molecular genetic technology

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