A Rapid PCR-Free Next-Generation Sequencing Method for the Detection of Copy Number Variations in Prenatal Samples
| Autor: | Na Hao, David S Cram, Yulin Jiang, Mengnan Xu, Chengkun Liu, Xiya Zhou, Xiangbin Chen, Juntao Liu, Qingwei Qi |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Microarray analysis techniques PCR-free libraries Paleontology Chromosome Prenatal diagnosis Computational biology 030105 genetics & heredity Biology General Biochemistry Genetics and Molecular Biology DNA sequencing Article 03 medical and health sciences genomic DNA 030104 developmental biology chromosome disorders copy number variation (CNV) Space and Planetary Science Gene duplication Amniocyte lcsh:Q Copy-number variation lcsh:Science rapid copy number variation sequencing (rCNV-seq) Ecology Evolution Behavior and Systematics |
| Zdroj: | Life Life, Vol 11, Iss 98, p 98 (2021) Volume 11 Issue 2 |
| ISSN: | 2075-1729 |
| Popis: | Next-generation sequencing (NGS) is emerging as a new method for the detection of clinically significant copy number variants (CNVs). In this study, we developed and validated rapid CNV-sequencing (rCNV-seq) for clinical application in prenatal diagnosis. Low-pass whole-genome sequencing was performed on PCR libraries prepared from amniocyte genomic DNA. From 10–40 ng of input DNA, PCR-free libraries consistently produced sequencing data with high unique read mapping ratios, low read redundancy, low coefficient of variation for all chromosomes and high genomic coverage. In validation studies, reliable and accurate CNV detection using PCR-free-based rCNV-seq was demonstrated for a range of common trisomies and sex chromosome aneuploidies as well as microdeletion and duplication syndromes. In reproducibility studies, CNV copy number and genomic intervals closely matched those defined by chromosome microarray analysis. Clinical testing of genomic DNA samples from 217 women referred for prenatal diagnosis identified eight samples (3.7%) with known chromosome disorders. We conclude that PCR-free-based rCNV-seq is a sensitive, specific, reproducible and efficient method that can be used in any NGS-based diagnostic laboratory for detection of clinically significant CNVs. |
| Databáze: | OpenAIRE |
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