scholarly journals Noninvasive prenatal diagnosis of hemophilia A by a haplotype-based approach using cell-free fetal DNA

BioTechniques ◽  
2020 ◽  
Vol 68 (3) ◽  
pp. 117-121
Author(s):  
Chao Chen ◽  
Jun Sun ◽  
Yun Yang ◽  
Lu Jiang ◽  
Fengyu Guo ◽  
...  
Keyword(s):  
Prenatal Diagnosis ◽  
Hemophilia A ◽  
Single Gene ◽  
Fetal Dna ◽  
Pathogenic Variants ◽  
Noninvasive Prenatal Diagnosis ◽  
Large Patient ◽  
Single Gene Disorders ◽  
Cell Free Fetal Dna ◽  
Consistency Rate

Aim: We aimed to demonstrate noninvasive prenatal diagnosis (NIPD) of hemophilia A (HA) using a haplotype-based approach. Methods: Two families at risk for HA were recruited for this study. First, maternal haplotypes associated with pathogenic variants were constructed using the genotypes of the mothers and probands. Then, fetal haplotypes were deduced using a maternal haplotype-assisted hidden Markov model. Finally, the NIPD results were further confirmed by invasive prenatal diagnosis. Results: Two fetal genotypes were successfully inferred, with one normal fetus and one carrier fetus. The NIPD results were confirmed by invasive prenatal diagnosis, with a 100% consistency rate. Conclusion: Our test has been shown to be accurate and reliable. With further validation in a large patient cohort, this haplotype-based approach could be feasible for the NIPD of HA and other X-linked single-gene disorders.

scholarly journals Applications of Cell-Free Fetal DNA in Maternal Serum

2012 ◽  
Vol 3 (2) ◽  
pp. 33-39 ◽  
Author(s):  
Saeid Ghorbian
Keyword(s):  
Prenatal Diagnosis ◽  
Single Gene ◽  
Late Pregnancy ◽  
Maternal Serum ◽  
Maternal Circulation ◽  
Fetal Dna ◽  
Noninvasive Prenatal Diagnosis ◽  
Cell Free Fetal Dna ◽  
Paternity Determination ◽  
Free Circulating Dna

ABSTRACT Cell-free fetal DNA (cffDNA) is available in the maternal circulation throughout pregnancy and can be used for noninvasive prenatal diagnosis including, determination of fetal sex, identification of specific single gene disorders, typing of fetal blood groups (RhD), paternity determination and potentially routine use for Down's syndrome (DS) testing of all pregnancies. I searched published literature on the PubMed and databases on Scopus interface systematically using keyword's cffDNA, noninvasive diagnosis, fetal DNA in the maternal serum. Reference lists from the papers were also searched. cffDNA representing only 3% of the total cell-free circulating DNA in early and rising to 12% in late pregnancy, clinical investigations has already demonstrated the potential advantage, such as improving safety, earlier diagnosis and comparative ease of testing using cffDNA technology. The discovery of cffDNA circulating in the maternal serum has opened the door to noninvasive prenatal diagnosis testing with novel clinical implications. How to cite this article Ghorbian S. Applications of Cell-Free Fetal DNA in Maternal Serum. Int J Infertility Fetal Med 2012;3(2):33-39.

scholarly journals Noninvasive prenatal diagnosis for Hemophilia A by haplotype-based approach using cell-free fetal DNA in maternal plasma

2019 ◽  
Author(s):  
Zhihui He ◽  
Chao Chen ◽  
Fengyu Guo ◽  
Lu Jiang ◽  
Yaping Zhu ◽  
...  
Keyword(s):  
Prenatal Diagnosis ◽  
Hemophilia A ◽  
Pathogenic Mutation ◽  
Maternal Plasma ◽  
Long Distance ◽  
High Coverage ◽  
Fetal Dna ◽  
Noninvasive Prenatal Diagnosis ◽  
Cell Free Fetal Dna ◽  
Haplotype A

AbstractObjectivesThis study was used to demonstrate the feasibility of haplotype-based noninvasive prenatal diagnosis for Hemophilia A.MethodsTwo families affected by Hemophilia A participated in our study. Maternal haplotypes associated with pathogenic mutation was built using targeted region genotypes of mother and the proband. Combined with maternal pathogenic haplotype, a Hidden Markov Model was constructed to deduce fetal haplotype using high-coverage targeted sequencing of maternal plasma. The presence of pathogenic haplotype in male indicated affected fetus, in female indicated carrier. Prenatal diagnosis was confirmed with amniocentesis by long distance PCR.ResultsThe haplotype-based noninvasive prenatal diagnosis was successfully performed in Hemophilia A. One fetus was identified to be normal, another fetus was identified as carrier, and the results were confirmed by amniocentesis .ConclusionOur research demonstrated the feasibility of noninvasive prenatal diagnosis for Hemophilia A by haplotype-based approach.

Maternal Plasma, as a Source for Fetal DNA, is Better than Fetal Cells for Noninvasive Prenatal Diagnosis of Single Gene Disorders

2012 ◽  
Vol 9 (1) ◽  
pp. S95
Author(s):  
Laura Hart ◽  
Janet Ober ◽  
Bhattacharya Anand ◽  
Yali Xiong ◽  
Owen Montgomery ◽  
...  
Keyword(s):  
Prenatal Diagnosis ◽  
Single Gene ◽  
Maternal Plasma ◽  
Fetal Cells ◽  
Fetal Dna ◽  
Noninvasive Prenatal Diagnosis ◽  
Single Gene Disorders ◽  
Better Than

scholarly journals Microsystem for Isolation of Fetal DNA from Maternal Plasma by Preparative Size Separation

2009 ◽  
Vol 55 (12) ◽  
pp. 2144-2152 ◽  
Author(s):  
Thomas Hahn ◽  
Klaus S Drese ◽  
Ciara K O'Sullivan
Keyword(s):  
Prenatal Diagnosis ◽  
High Volume ◽  
Maternal Plasma ◽  
Chromosomal Anomalies ◽  
Size Separation ◽  
Large Samples ◽  
Fetal Dna ◽  
Noninvasive Prenatal Diagnosis ◽  
Cell Free Fetal Dna ◽  
Invasive Techniques

Abstract Background: Routine prenatal diagnosis of chromosomal anomalies is based on invasive procedures, which carry a risk of approximately 1%–2% for loss of pregnancy. An alternative to these inherently invasive techniques is to isolate fetal DNA circulating in the pregnant mother’s plasma. Free fetal DNA circulates in maternal plasma primarily as fragments of lengths <500 bp, with a majority being <300 bp. Separating these fragments by size facilitates an increase in the ratio of fetal to maternal DNA. Methods: We describe our development of a microsystem for the enrichment and isolation of cell-free fetal DNA from maternal plasma. The first step involves a high-volume extraction from large samples of maternal plasma. The resulting 80-μL eluate is introduced into a polymeric microsystem within which DNA is trapped and preconcentrated. This step is followed by a transient isotachophoresis step in which the sample stacks within a neighboring channel for subsequent size separation and is recovered via an outlet at the end of the channel. Results: Recovered fractions of fetal DNA were concentrated 4–8 times over those in preconcentration samples. With plasma samples from pregnant women, we detected the fetal SRY gene (sex determining region Y) exclusively in the fragment fraction of <500 bp, whereas a LEP gene (leptin) fragment was detected in both the shorter and longer recovery fractions. Conclusions: The microdevice we have described has the potential to open new perspectives in noninvasive prenatal diagnosis by facilitating the isolation of fetal DNA from maternal plasma in an integrated, inexpensive, and easy-to-use microsystem.

scholarly journals Noninvasive Prenatal Diagnosis of Single-Gene Disorders by Use of Droplet Digital PCR

2018 ◽  
Vol 64 (2) ◽  
pp. 336-345 ◽  
Author(s):  
Joan Camunas-Soler ◽  
Hojae Lee ◽  
Louanne Hudgins ◽  
Susan R Hintz ◽  
Yair J Blumenfeld ◽  
...  
Keyword(s):  
At Risk ◽  
Prenatal Diagnosis ◽  
Metabolic Disorders ◽  
Single Gene ◽  
Digital Pcr ◽  
Maternal Plasma ◽  
Single Nucleotide ◽  
Noninvasive Prenatal Diagnosis ◽  
Single Gene Disorders ◽  
Fetal Fraction

Abstract BACKGROUND Prenatal diagnosis in pregnancies at risk of single-gene disorders is currently performed using invasive methods such as chorionic villus sampling and amniocentesis. This is in contrast with screening for common aneuploidies, for which noninvasive methods with a single maternal blood sample have become standard clinical practice. METHODS We developed a protocol for noninvasive prenatal diagnosis of inherited single-gene disorders using droplet digital PCR from circulating cell-free DNA (cfDNA) in maternal plasma. First, the amount of cfDNA and fetal fraction is determined using a panel of TaqMan assays targeting high-variability single-nucleotide polymorphisms. Second, the ratio of healthy and diseased alleles in maternal plasma is quantified using TaqMan assays targeting the mutations carried by the parents. Two validation approaches of the mutation assay are presented. RESULTS We collected blood samples from 9 pregnancies at risk for different single-gene disorders, including common conditions and rare metabolic disorders. We measured cases at risk of hemophilia, ornithine transcarbamylase deficiency, cystic fibrosis, β-thalassemia, mevalonate kinase deficiency, acetylcholine receptor deficiency, and DFNB1 nonsyndromic hearing loss. We correctly differentiated affected and unaffected pregnancies (2 affected, 7 unaffected), confirmed by neonatal testing. We successfully measured an affected pregnancy as early as week 11 and with a fetal fraction as low as 3.7% (0.3). CONCLUSIONS Our method detects single-nucleotide mutations of autosomal recessive diseases as early as the first trimester of pregnancy. This is of importance for metabolic disorders in which early diagnosis can affect management of the disease and reduce complications and anxiety related to invasive testing.

scholarly journals Bayesian-based noninvasive prenatal diagnosis of single-gene disorders

2019 ◽  
Vol 29 (3) ◽  
pp. 428-438 ◽  
Author(s):  
Tom Rabinowitz ◽  
Avital Polsky ◽  
David Golan ◽  
Artem Danilevsky ◽  
Guy Shapira ◽  
...  
Keyword(s):  
Prenatal Diagnosis ◽  
Single Gene ◽  
Noninvasive Prenatal Diagnosis ◽  
Single Gene Disorders

scholarly journals Noninvasive Prenatal Detection of Fetal Trisomy 18 by Epigenetic Allelic Ratio Analysis in Maternal Plasma: Theoretical and Empirical Considerations

2006 ◽  
Vol 52 (12) ◽  
pp. 2194-2202 ◽  
Author(s):  
Yu K Tong ◽  
Chunming Ding ◽  
Rossa WK Chiu ◽  
Ageliki Gerovassili ◽  
Stephen SC Chim ◽  
...  
Keyword(s):  
Prenatal Diagnosis ◽  
Maternal Plasma ◽  
Trisomy 18 ◽  
Ratio Analysis ◽  
Dna Mixtures ◽  
Allelic Ratio ◽  
Fetal Dna ◽  
Noninvasive Prenatal Diagnosis ◽  
Cell Free Fetal Dna ◽  
Chromosomal Aneuploidies

Abstract Background: The discovery of cell-free fetal DNA in maternal plasma has opened up new possibilities for noninvasive prenatal diagnosis. However, the use of maternal plasma fetal DNA for the direct detection of fetal chromosomal aneuploidies has not been reported. We postulate that the aneuploidy status of a fetus could be revealed by an epigenetic allelic ratio approach, i.e., by analyzing the allelic ratio of a single-base variation present within DNA molecules exhibiting a placental-specific epigenetic signature in maternal plasma. Methods: Placental-derived fetal-specific unmethylated maspin (SERPINB5) promoter sequences on human chromosome 18 were detectable in placental–maternal DNA mixtures and in maternal plasma by bisulfite modification followed by methylation-specific PCR (MSP) and primer extension. The ratios between the extension products of the 2 alleles were calculated for heterozygous placentas, placental–maternal blood cell DNA mixtures, and maternal plasma samples. The allelic ratios were compared between pregnancies carrying trisomy 18 and euploid fetuses. Results: The epigenetic allelic ratios of all tested trisomy 18 samples deviated from the reference range obtained from euploid samples (placental DNA, 1.135 to 2.052; placental–maternal DNA mixtures, 1.170 to 1.985; maternal plasma, 0.330 to 3.044; without skew correction on the raw mass spectrometric data). A theoretical model was established and validated that predicted that a minimum of 200 copies of genomic DNA after bisulfite conversion were required for distinguishing euploid and aneuploid fetuses with confidence. Conclusion: Epigenetic allelic ratio analysis of maternal plasma DNA represents a promising approach for noninvasive prenatal diagnosis of fetal chromosomal aneuploidies.

scholarly journals Noninvasive Prenatal Diagnosis from Maternal Blood: Finally Available after 20 Years of Research

2013 ◽  
Vol 7 (4) ◽  
pp. 440-442
Author(s):  
Wolfgang Holzgreve
Keyword(s):  
Prenatal Diagnosis ◽  
Single Gene ◽  
Chorionic Villus ◽  
Maternal Blood ◽  
Chorionic Villus Sampling ◽  
Clinical Use ◽  
Maternal Circulation ◽  
Isolated Cells ◽  
Noninvasive Prenatal Diagnosis ◽  
Cell Free Fetal Dna

ABSTRACT Since all prenatal invasive procedures, such as amniocentesis and chorionic villus sampling carry a small risk for the pregnant woman and a risk to induce the loss of a pregnancy of up to 1%, there have been efforts now for at least a quarter of a century to develop a noninvasive method from the blood of pregnant women. First there was a considerable effort to isolate fetal cells from maternal circulation, and these techniques were carefully evaluated in a NIH-sponsored study of a few US American centers and ours in Basel/Switzerland. It turned out; however, that interphase fluorescence to identify fetal aneuploidies from these isolated cells was not reliable enough for clinical use. The breakthrough came with the recognition of the group by D Lo et al; who showed for the first time that cell-free fetal DNA in maternal plasma and serum can be used reliably for prenatal diagnosis. One of the first successful applications was the detection of the fetal Rhesus factor around 11 weeks of gestation in pregnancies of Rhesus-negative mothers. The Sequenom Company in San Diego, USA, which acquired the patent of D Lo et al on the use of cell free DNA and ours on size separation of fetal vs maternal DNA subsequently showed in large series that the noninvasive prenatal diagnosis of fetal trisomy 21 from maternal blood by massive parallel sequencing has an accuracy around 99%, and currently up to 100,000 cases have been investigated already in different laboratories. Also the noninvasive prenatal diagnosis of trisomies 18 and 13 is possible, and an increasing amount of single gene anomalies will be diagnosable in the future noninvasively. The whole development of noninvasive prenatal diagnosis is appositive example that long-term research pays-off to bring a concept from the first steps finally into clinical use. How to cite this article Holzgreve W. Noninvasive Prenatal Diagnosis from Maternal Blood: Finally Available after 20 Years of Research. Donald School J Ultrasound Obstet Gynecol 2013;7(4):440-442.

scholarly journals Recent Advances in Fetal Nucleic Acids in Maternal Plasma

2005 ◽  
Vol 53 (3) ◽  
pp. 293-296 ◽  
Author(s):  
Y.M. Dennis Lo
Keyword(s):  
Prenatal Diagnosis ◽  
First Trimester ◽  
Thalassemia Major ◽  
Maternal Plasma ◽  
Fetal Dna ◽  
Noninvasive Prenatal Diagnosis ◽  
Cell Free Fetal Dna ◽  
Circulating Fetal Dna ◽  
Circulating Rna ◽  
Near Future

The discovery of cell-free fetal DNA in maternal plasma in 1997 has opened up new possibilities for noninvasive prenatal diagnosis. Circulating fetal DNA molecules have been detected in maternal plasma from the first trimester onwards and can be robustly detected using a variety of molecular methods. This approach has been used for the prenatal investigation of sex-linked diseases, fetal RhD status, and prenatal exclusion of β-thalassemia major. Recently, fetal RNA has also been found in maternal plasma. Such fetal RNA has been shown to originate from the placenta and to be remarkably stable. The use of microarray-based approaches has made it feasible to rapidly generate new circulating RNA markers. It is hoped that further developments in this field will make the routine and widespread practice of noninvasive nucleic acid-based prenatal diagnosis for common pregnancy-associated disorders feasible in the near future.

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