Noninvasive fetal trisomy detection by multiplexed semiconductor sequencing: a barcoding analysis strategy

Noninvasive prenatal detection of fetal chromosomal aneuploidies by high-throughput next-generation sequencing proves to be accurate and sensitive. Currently, most of the data analysis methods involve a Z-score test based on the reference distribution of at least dozens of normal samples. This is not only costly but also time consuming. Moreover, as the experimental condition varies between every single run, noises cannot be eliminated and will skew the results. In order to overcome these drawbacks, we have proposed a new analytical strategy based on the multiplex barcoding sequencing of both normal and unknown samples in a single run on Ion Torrent PGM. In this method, only one normal sample is required. By applying this method to 13 single runs with a total number of 44 samples, we achieved the sensitivity and specificity of 100 and 95.181% for T13 detection, 100 and 100% for T18 detection, 90 and 100% for T21 detection, respectively.