Clinical Validation of Novel Chip-Based Digital PCR Platform for Fetal Aneuploidies Screening.

Anna Nykel,Rafał Woźniak,Agnieszka Gach

doi:10.3390/diagnostics11071131

Abstract

Fetal aneuploidy is routinely diagnosed by karyotyping. The development of techniques for rapid aneuploidy detection based on the amplification reaction allows cheaper and rapid diagnosis. However, the currently available solutions have limitations. We tested a novel approach as a diagnostic tool in clinical practice. The objective of this study was to provide a clinical performance of the sensitivity and specificity of a novel chip-based digital PCR approach for fetal aneuploidy screening. The study was conducted in 505 pregnant women with increased risk for fetal aneuploidy undergoing invasive prenatal diagnostics. DNA extracted from amniotic fluid or CVS was analyzed for the copy number of chromosomes 13, 18, 21, X, and Y using a new chip-based solution. Performance was assessed by comparing results with findings from karyotyping. Aneuploidy was confirmed in 65/505 cases positive for trisomy 21, 30/505 cases positive for trisomy 18, 14/505 cases positive for trisomy 13 and 21/505 with SCAs. Moreover, 2 cases with triploidy and 2 cases with confirmed mosaicisms of 21 and X chromosomes were detected. Clinical sensitivity and specificity within this study was determined at 100% for T21 (95% CI, 99.26–100%), T18 (95% CI, 99.26–100%), and T13 (95% CI, 99.26–100%). Chip-based digital PCR provides equally high sensitivity and specificity in rapid aneuploidy screening and can be implemented into routine prenatal diagnostics.

Highlights

Chromosome abnormalities cause birth defects and are present in approximately3–5% of all clinically diagnosed pregnancies [1]
110 cases were classified as the most common autosomal trisomies (T13, T18, T21) which cases were classified as the most common autosomal trisomies (T13, T18, T21) which corcorresponds to 88.70% of all abnormal results, 13 cases tested positive for sex chromosomal aneuploidies (SCAs) (10.40% of responds to 88.70% of all abnormal results, 13 cases tested positive for SCAs (10.40% of abnormal), and in 2 cases male triploidy was diagnosed (1.60% of abnormal)
T21 male euploid female euploid female. This is the first study documenting the clinical effectiveness of the novel approach using QuantStudio 3D Digital PCR platform for the detection of the most common fetal aneuploidies, such as trisomy 21, trisomy 18, trisomy 13, and SCAs

Summary

Introduction

Chromosome abnormalities cause birth defects and are present in approximately3–5% of all clinically diagnosed pregnancies [1]. Chromosome abnormalities cause birth defects and are present in approximately. The most common numerical autosomal abnormalities are trisomy 21 (Down syndrome), trisomy 18 (Edwards syndrome), and trisomy 13 (Patau syndrome) [1]. Birth defects can be caused by X or Y chromosome abnormality, known as sex chromosomal aneuploidies (SCAs). The aim of standard prenatal care is the identification of women with increased risk of fetal aneuploidies using biochemical markers and ultrasound markers in the first and second trimester [2,3]. The gold standard for prenatal diagnosis is conventional cytogenetic analyses. Obtaining a karyotyping result based on cell culture of amniotic fluid or chorionic villus samples require approximately two weeks. A rapid and precise molecular method for aneuploidy detection is desirable. The most popular rapid diagnostics is based on real-time

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