Non‐invasive prenatal diagnostic testing for β‐thalassaemia using cell‐free fetal DNA and next generation sequencing

• DOI: 10.1002/pd.4536
• Publication Date: 2014-11-14T15:11:12Z
• Authors: Li Xiong; Angela N. Barrett; Rui Hua; Tuan Zea Tan; Sherry Sze Yee Ho; Jerry K. Y. Chan; Mei Zhong; Mahesh Choolani
• Keywords: Cell-free fetal DNA; Amplicon; Compound heterozygosity

Abstract Objective To develop an accurate non‐invasive prenatal test using next generation sequencing (NGS) for HbE and the four most common β‐thalassaemia mutations found in South East Asia (namely −28A > G, CD17A T, CD41/42(−TTCT) IVS‐II‐654C T). Methods Cell‐free DNA was extracted from maternal plasma 83 families where both parents were carriers of mutation or one mutations. Overlapping PCR amplicons covering each generated, pooled sequenced Illumina MiSeq. Fastq files analysed to detect inheritance paternal mutation. Results In two cases fathers compound heterozygotes fetus correctly diagnosed as having inherited 35/85 cases, not detected, 50/85 it classified inherited. Overall sensitivity detection 100% (95% CI: 92.4–100%), specificity 92.1% 79.2–97.3%). Conclusion We demonstrated that NGS can be readily achieved with high specificity, removing need invasive 50% pregnancies at risk a thalassaemia father mother carry different © 2014 John Wiley & Sons, Ltd.