Noninvasive prenatal detection of chromosomal aneuploidies using different next generation sequencing strategies and algorithms

Abstract

Our results confirm previous reports that massively parallel sequencing of cell-free fetal DNA allows the reliably noninvasive detection of trisomy 21 from maternal blood with the potential to enhance test selectivity and specificity by bioinformatic means. According to our preliminary results, targeted sequencing might be an alternative strategy to detect chromosomal aneuploidies besides trisomy 21.

Diagnostic accuracy of random massively parallel sequencing for non-invasive prenatal detection of common autosomal aneuploidies: a collaborative study in Europe

“…Seven to 10 mL of peripheral venous blood was taken from participating pregnant women prior to invasive procedures using either conventional EDTA blood collection tubes (BCT) (Sarstedt) (Centers in Berlin and Munich only) or Cell‐Free DNA™ BCT (Streck Innovations) (all centers except Munich). EDTA blood samples were processed by the centers for extraction of plasma as previously described . The cell‐free plasma samples were stored at −20 °C and subsequently shipped (cooled) to LifeCodexx AG and GATC Biotech AG.…”

“…Here, we report the results of a collaborative European clinical study, in which five clinical centers in Germany and Switzerland were involved. The primary aim was to validate the diagnostic accuracy of an established noninvasive laboratory test based on random MPS for detecting fetal trisomy 21 in these populations, as a continuation of a recently published proof‐of‐concept study . Furthermore, we compared the accuracy of two bioinformatic analysis pipelines – DAP.21 focused on detection of trisomy 21 only, and DAP.plus based on an algorithm using guanine‐cytosine (GC) normalization – along with their ability to detect trisomies 13 and 18, in addition to trisomy 21.…”

Hypermethylated ERG as a cell-free fetal DNA biomarker for non-invasive prenatal testing of Down syndrome

“…Next generation sequencing (NGS) is a new approach for analyzing cell-free DNA in maternal plasma by massive parallel sequencing, which has been successfully applied to NIPT for fetal aneuploidy. However, NGS requires professional laboratory technicians and expensive sequencing equipment, which makes the testing expensive and may decrease utilization [12]. In this study, we introduced a qPCR method for rapid screening trisomy 21 by analyzing cell-free DNA in maternal plasma, which is both specific and cost effective.…”

Systematic review of noninvasive prenatal diagnosis for abnormal chromosome genetic diseases using free fetal DNA in maternal plasma

“…Karyotyping or fluorescence in situ hybridization (FISH) (the traditional diagnosis method) is conducted to determine the presence of chromosome abnormalities in the fetus in the high-risk gravida (Qu et al, 2013). A limitation of this traditional diagnosis method is the false positive rate of 5% in Down's syndrome screening (Alldred, 2012), and karyotyping as well as FISH are invasive diagnosis methods that unnecessarily cause high-risk gravidas to suffer from the pain of chorionic villus, umbilical cord blood collection, and amniotic fluid puncture (Stumm et al, 2012).…”