Detection of paternal alleles in maternal plasma for non-invasive prenatal diagnosis of β-thalassemia: A feasibility study in southern Chinese

Abstract Background: To limit risks of miscarriages associated with invasive procedures of current prenatal diagnosis practice, we aim to develop a personalized medicine-based protocol for non-invasive prenatal diagnosis (NIPD) of monogenic disorders relying on the detection of paternally inherited mutations in maternal blood using droplet digital PCR (ddPCR). Methods: This study included four couples at risk of transmitting paternal neurofibromatosis type 1 (NF1) mutations and four couples at risk of transmitting compound heterozygous CFTR mutations. NIPD was performed between 8 and 15 weeks of gestation, in parallel to conventional invasive diagnosis. We designed specific hydrolysis probes to detect the paternal mutation and to assess the presence of cell-free fetal DNA by ddPCR. Analytical performances of each assay were determined from paternal sample, an then fetal genotype was inferred from maternal plasma sample. Results: Presence or absence of the paternal mutant allele was correctly determined in all the studied plasma DNA samples. Conclusions: We report an NIPD protocol suitable for implementation in an experienced laboratory of molecular genetics. Our proof-of-principle results point out a high accuracy for early detection of paternal NF1 and CFTR mutations in cell-free DNA, and open new perspectives for extending the technology to NIPD of many other monogenic diseases.

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Use of maternal plasma for non-invasive prenatal diagnosis of fetal ABO genotypes

Clinical Chemistry and Laboratory Medicine (CCLM) ◽

10.1515/cclm.2007.193 ◽

2007 ◽

Vol 45 (8) ◽

Cited By ~ 3

Author(s):

Jin-Lai Meng ◽

Xie-Tong Wang ◽

Yu Wang ◽

Ya-Fei Yue ◽

Xin Wang ◽

...

Keyword(s):

Prenatal Diagnosis ◽

Maternal Plasma ◽

Non Invasive ◽

Abo Genotypes

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The SAFE project: towards non-invasive prenatal diagnosis

Biochemical Society Transactions ◽

10.1042/bst0370460 ◽

2009 ◽

Vol 37 (2) ◽

pp. 460-465 ◽

Cited By ~ 24

Author(s):

Deborah G. Maddocks ◽

Medhat S. Alberry ◽

George Attilakos ◽

Tracey E. Madgett ◽

Kin Choi ◽

...

Keyword(s):

Prenatal Diagnosis ◽

Large Scale ◽

Single Gene ◽

Cost Effective ◽

Maternal Plasma ◽

Molecular Techniques ◽

Maternal Circulation ◽

Haemolytic Disease ◽

Non Invasive ◽

New Biomarkers

After the revolutionary detection of ffDNA (free fetal DNA) in maternal circulation by real-time PCR in 1997 and advances in molecular techniques, NIPD (non-invasive prenatal diagnosis) is now a clinical reality. Non-invasive diagnosis using ffDNA has been implemented, allowing the detection of paternally inherited alleles, sex-linked conditions and some single-gene disorders and is a viable indicator of predisposition to certain obstetric complications [e.g. PET (pre-eclampsia)]. To date, the major use of ffDNA genotyping in the clinic has been for the non-invasive detection of the pregnancies that are at risk of HDFN (haemolytic disease of the fetus and newborn). This has seen numerous clinical services arising across Europe and many large-scale NIPD genotyping studies taking place using maternal plasma. Because of the interest in performing NIPD and the speed at which the research in this area was developing, the SAFE (Special Non-Invasive Advances in Fetal and Neonatal Evaluation) NoE (Network of Excellence) was founded. The SAFE project was set up to implement routine, cost-effective NIPD and neonatal screening through the creation of long-term partnerships within and beyond the European Community and has played a major role in the standardization of non-invasive RHD genotyping. Other research using ffDNA has focused on the amount of ffDNA present in the maternal circulation, with a view to pre-empting various complications of pregnancy. One of the key areas of interest in the non-invasive arena is the prenatal detection of aneuploid pregnancies, particularly Down's syndrome. Owing to the high maternal DNA background, detection of ffDNA from maternal plasma is very difficult; consequently, research in this area is now more focused on ffRNA to produce new biomarkers.

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