The Performance of Whole Genome Amplification Methods and Next-Generation Sequencing for Pre-Implantation Genetic Diagnosis of Chromosomal Abnormalities

Reliable and accurate pre-implantation genetic diagnosis （PGD） of patient＇s embryos by next-generation sequencing （NGS） is dependent on efficient whole genome amplification （WGA） of a representative biopsy sample. However, the performance of the current state of the art WGA methods has not been eval...

Full description

Saved in:
Bibliographic Details
Published in:Journal of genetics and genomics 2015-04, Vol.42 (4), p.151-159
Main Authors: Li, Na, Wang, Li, Wang, Hui, Ma, Minyue, Wang, Xiaohong, Li, Yi, Zhang, Wenke, Zhang, Jianguang, Cram, David S., Yao, Yuanqing
Format: Article
Language:English
Subjects:
Chromosome Aberrations - embryology
Copy number variation
DNA Copy Number Variations - genetics
GA系统
Genome, Human - genetics
High-Throughput Nucleotide Sequencing - methods
Humans
Next-generation sequencing
Polymerase Chain Reaction - methods
Pre-implantation genetic diagnosis
Preimplantation Diagnosis - methods
Sensitivity and Specificity
Sequence Analysis, DNA - methods
Single cells
Single-Cell Analysis - methods
Whole genome amplification
全基因组扩增
基因诊断
性能
扩增方法
染色体异常
测序
胚胎植入
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Reliable and accurate pre-implantation genetic diagnosis （PGD） of patient＇s embryos by next-generation sequencing （NGS） is dependent on efficient whole genome amplification （WGA） of a representative biopsy sample. However, the performance of the current state of the art WGA methods has not been evaluated for sequencing. Using low template DNA （15 pg） and single cells, we showed that the two PCR-based WGA systems SurePlex and MALBAC are superior to the REPLI-g WGA multiple displacement amplification （MDA） system in terms of consistent and reproducible genome coverage and sequence bias across the 24 chromosomes, allowing better normalization of test to reference sequencing data. When copy number variation sequencing （CNV-Seq） was applied to single cell WGA products derived by either SurePlex or MALBAC amplification, we showed that known disease CNVs in the range of 3-15 Mb could be reliably and accurately detected at the correct genomic positions. These findings indicate that our CNV-Seq pipeline incorporating either SurePlex or MALBAC as the key initial WGA step is a powerful methodology for clinical PGD to identify euploid embryos in a patient＇s cohort for uterine transplantation,
ISSN:1673-8527
DOI:10.1016/j.jgg.2015.03.001