OmniPlex—a new QF‐PCR assay for prenatal diagnosis of common aneuploidies based on evaluation of the heterozygosity of short tandem repeat loci in the Czech population

Abstract

The aim of our study was to assess the utility of commonly used multiplex assays of short tandem repeat markers used for quantitative fluorescent polymerase chain reaction (QF-PCR) for prenatal rapid aneuploidy detection (RAD) in routine prenatal diagnosis in the Czech population. Two previously published RAD multiplexes (2M test) were tested on 2906 local prenatal samples and used to calculate the rates of heterozygosity within this population. Most of the markers used in both multiplexes were highly informative. However, some had little utility, either due to a low heterozygosity rate (D21S499, D18S978 and P39) or because they were difficult to evaluate (DXS1283E). After evaluation of the 2M test results, a new multiplex assay (OmniPlex) was designed, developed and tested on 960 samples. This new assay was evaluated for heterozygosity rates and for the probability of having two or more informative markers on each chromosome. OmniPlex assay significantly improved the QF-PCR methodology for rapid prenatal aneuploidy detection in the Czech population. Based on detected heterozygosity of markers used for QF-PCR in this population, OmniPlex is a robust assay for the detection of chromosomes 13, 18, 21, X and Y in a single reaction.