Development and validation of a next-generation sequencing-based protocol for 24-chromosome aneuploidy screening of embryos.
Fertil Steril
; 101(5): 1375-82, 2014 May.
Article
em En
| MEDLINE
| ID: mdl-24613537
ABSTRACT
OBJECTIVE:
To validate a next-generation sequencing (NGS)-based method for 24-chromosome aneuploidy screening and to investigate its applicability to preimplantation genetic screening (PGS).DESIGN:
Retrospective blinded study.SETTING:
Reference laboratory. PATIENT(S) Karyotypically defined chromosomally abnormal single cells and whole-genome amplification (WGA) products, previously analyzed by array comparative genomic hybridization (array-CGH), selected from 68 clinical PGS cycles with embryos biopsied at cleavage stage. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Consistency of NGS-based diagnosis of aneuploidy compared with either conventional karyotyping of single cells or array-CGH diagnoses of single blastomeres. RESULT(S) Eighteen single cells and 190 WGA products from single blastomeres, were blindly evaluated with the NGS-based protocol. In total, 4,992 chromosomes were assessed, 402 of which carried a copy number imbalance. NGS specificity for aneuploidy call (consistency of chromosome copy number assignment) was 99.98% (95% confidence interval [CI] 99.88%-100%) with a sensitivity of 100% (95% CI 99.08%-100%). NGS specificity for aneuploid embryo call (24-chromosome diagnosis consistency) was 100% (95% CI 94.59%-100%) with a sensitivity of 100% (95% CI 97.39%-100%). CONCLUSION(S) This is the first study reporting extensive preclinical validation and accuracy assessment of NGS-based comprehensive aneuploidy screening on single cells. Given the high level of consistency with an established methodology, such as array-CGH, NGS has demonstrated a robust high-throughput methodology ready for clinical application in reproductive medicine, with potential advantages of reduced costs and enhanced precision.Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Diagnóstico Pré-Natal
/
Aneuploidia
/
Hibridização de Ácido Nucleico
Tipo de estudo:
Diagnostic_studies
/
Guideline
/
Observational_studies
/
Screening_studies
Limite:
Adult
/
Female
/
Humans
/
Middle aged
/
Pregnancy
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article