domingo, 5 de octubre de 2014

Development of DNA Confirmatory and High-Risk Diagnostic Testing for Newborns Using Targeted Next-Generation DNA Sequencing : Genetics in Medicine : Nature Publishing Group

Algorithm and workflow for next-generation sequencing (NGS)-based newborn confirmatory and diagnostic testing. (a) Exome (whole-exome sequencing, WES) and 126-gene panel (NBDx) analysis using in silico gene filters. A subset of the exo…

Development of DNA Confirmatory and High-Risk Diagnostic Testing for Newborns Using Targeted Next-Generation DNA Sequencing

Genetics in Medicine

(2014)

doi:10.1038/gim.2014.117

Received: 18 March 2014
Accepted: 31 July 2014
Published online: 25 September 2014

Abstract

Purpose:

Genetic testing is routinely used for second-tier confirmation of newborn sequencing results to rule out false positives and to confirm diagnoses in newborns undergoing inpatient and outpatient care. We developed a targeted next-generation sequencing panel coupled with a variant processing pipeline and demonstrated utility and performance benchmarks across multiple newborn disease presentations in a retrospective clinical study.

Methods:

The test utilizes an in silico gene filter that focuses directly on 126 genes related to newborn screening diseases and is applied to the exome or a next-generation sequencing panel called NBDx. NBDx targets the 126 genes and additional newborn-specific disorders. It integrates DNA isolation from minimally invasive biological specimens, targeted next-generation screening, and rapid characterization of genetic variation.

Results:

We report a rapid parallel processing of 8 to 20 cases within 105 hours with high coverage on our NBDx panel. Analytical sensitivity of 99.8% was observed across known mutation hotspots. Concordance calls with or without clinical summaries were 94% and 75%, respectively.

Conclusion:

Rapid, automated targeted next-generation sequencing and analysis are practical in newborns for second-tier confirmation and neonatal intensive care unit diagnoses, laying a foundation for future primary DNA-based molecular screening of additional disorders and improving existing molecular testing options for newborns.

Genet Med advance online publication 25 September 2014

View full text