Dello Russo C., Cesta A., Longo S., Barone M.A., Cima A., Mesoraca A., Sparacino D., Viola A., Giorlandino C.

Validation of Extensive Next-Generation Sequencing Method for Monogenic Disorder Analysis on Cell-Free Fetal DNA: Noninvasive Prenatal Diagnosis

J Mol Diagn, 2019 Jul 21


During pregnancy, a percentage of the cell-free DNA circulating in the maternal blood is represented by the cell-free fetal DNA (cffDNA), constituting an accessible source for noninvasive prenatal genetic screening. The coexistence of the maternal DNA, the dominant fraction of cell-free DNA, together with the cffDNA component and the scarcity of the cffDNA itself make applying traditional methods of genetics and molecular biology impossible.

Next-generation sequencing methods are widely used to study fetal aneuploidies. However, in monogenic disorders, there have been relatively few studies that analyzed single mutations. We present a method for the analysis of an extended group of gene variants associated with recessive and dominant autosomal disorders using next-generation sequencing.

The proposed test should allow a complete analysis of common genetic disorders and pathogen-associated variants for diagnostic use. The analysis of cffDNA for single gene disorders may replace invasive prenatal diagnosis methods, associated with the risk of spontaneous abortion and psychological stress for patients. The proposed test should assess reproductive risk for both genetic family disorders and de novo occurrences of the disease. The application of this method to a case of beta-thalassemia is also discussed.

Mesoraca A., Margiotti K., Dello Russo C., Cesta A., Cima A., Longo S., Barone M. A., Viola. A, Sparacino D., Giorlandino C.

Cell-free DNA screening for aneuploidies in 7113 pregnancies: single Italian centre study

Genet Res (Camb), 2020 Jun 16


Non-invasive prenatal testing (NIPT) using cell-free foetal DNA has been widely accepted in recent years for detecting common foetal chromosome aneuploidies, such as trisomies 13, 18 and 21, and sex chromosome aneuploidies.

In this study, the practical clinical performance of our foetal DNA testing was evaluated for analysing all chromosome aberrations among 7113 pregnancies in Italy. In this study, NIPT showed 100% sensitivity and 99.9% specificity for trisomies 13, 18 and 21.

Margiotti K., Cesta A., Dello Russo C., Cima A., Barone M.A., Viola A., Sparacino D., Mesoraca A., Giorlandino C.

Cell-free DNA screening for sex chromosomal aneuploidies in 9985 pregnancies: Italian single experience

BMC Res Notes, 2020 Mar 18


In this study, the clinical performance of our fetal DNA testing was investigated by analyzing the sex chromosome ane- uploidy aberrations among 9985 pregnancies. The study was a retrospective analysis of collected NIPT data from the Ion S5 next-generation sequencing (NGS) platform obtained from Altamedica Medical Centre of Rome.

In conclusion, the present results confirm that NIPT is a potential method for SCA screening


Orhant L., Rondeau S., Vasson A., Anselem O., Goffinet F., Allach El Khattabi L., Leturcq F., Vidaud D., Bienvenu T., Tsatsaris V., Nectoux,

Droplet digital PCR, a new approach to analyze fetal DNA from maternal blood: application to the determination of fetal RHD genotype

Ann Biol Clin. 2016 Jun 1

Liao G.J., Gronowski A.M., Zhao Z.

Non-invasive prenatal testing using cell-free fetal DNA in maternal circulation

Clin. Chim. Acta. 2014 Jan 20

El Khattabi L.A., Rouillac-Le Sciellour C., Le Tessier D., Luscan A., Coustier A., Porcher R.

Could Digital PCR Be an Alternative as a Non-Invasive Prenatal Test for Trisomy 21: A Proof of Concept Study

PLoS One. 2016 May 11

Jeon Y.J., Zhou Y., Li Y., Guo Q., Chen J.

The feasibility study of non-invasive fetal trisomy 18 and 21 detection with semiconductor sequencing platform

PLoS One. 2014 Oct 20

Karakas B., Qubbaj W., Al-Hassan S., Coskun S.

Noninvasive Digital Detection of Fetal DNA in Plasma of 4-Week-Pregnant Women following In Vitro Fertilization and Embryo Transfer

PLoS One. 2015 May 13