Automated workflow for somatic and Germline next generation sequencing analysis in routine clinical cancer diagnostics

Lucia Anna Muscarella, Federico Pio Fabrizio, Maria De Bonis, Maria Teresa Mancini, Teresa Balsamo, Paolo Graziano, Flavia Centra, Angelo Sparaneo, Domenico Trombetta, Antonio Bonfitto, Vito Scagliusi, Pietro Larizza, Ettore Domenico Capoluongo, Vito Michele Fazio

Research output: Contribution to journalArticle

Abstract

Thanks to personalized medicine trends and collaborations between industry, clinical research groups and regulatory agencies, next generation sequencing (NGS) is turning into a common practice faster than one could have originally expected. When considering clinical applications of NGS in oncology, a rapid workflow for DNA extraction from formalin-fixed paraffin-embedded (FFPE) tissue samples, as well as producing high quality library preparation, can be real challenges. Here we consider these targets and how applying effective automation technology to NGS workflows may help improve yield, timing and quality-control. We firstly evaluated DNA recovery from archived FFPE blocks from three different manual extraction methods and two automated extraction workstations. The workflow was then implemented to somatic (lung/colon panel) and germline (BRCA1/2) library preparation for NGS analysis exploiting two automated workstations. All commercial kits gave good results in terms of DNA yield and quality. On the other hand, the automated workstation workflow has been proven to be a valid automatic extraction system to obtain high quality DNA suitable for NGS analysis (lung/colon Ampli-seq panel). Moreover, it can be efficiently integrated with an open liquid handling platform to provide high-quality libraries from germline DNA with more reproducibility and high coverage for targeted sequences in less time (BRCA1/2). The introduction of automation in routine workflow leads to an improvement of NGS standardization and increased scale up of sample preparations, reducing labor and timing, with optimization of reagents and management.

Original languageEnglish
Article number1691
JournalCancers
Volume11
Issue number11
DOIs
Publication statusPublished - Nov 2019

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Workflow
Automation
DNA
Neoplasms
Paraffin
Formaldehyde
Libraries
Colon
Lung
Precision Medicine
Gene Library
Quality Control
Industry
Technology
Research

Keywords

  • Automation
  • DNA extraction
  • FFPE
  • Library preparation
  • NGS

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Automated workflow for somatic and Germline next generation sequencing analysis in routine clinical cancer diagnostics. / Muscarella, Lucia Anna; Fabrizio, Federico Pio; De Bonis, Maria; Mancini, Maria Teresa; Balsamo, Teresa; Graziano, Paolo; Centra, Flavia; Sparaneo, Angelo; Trombetta, Domenico; Bonfitto, Antonio; Scagliusi, Vito; Larizza, Pietro; Capoluongo, Ettore Domenico; Fazio, Vito Michele.

In: Cancers, Vol. 11, No. 11, 1691, 11.2019.

Research output: Contribution to journalArticle

Muscarella, Lucia Anna ; Fabrizio, Federico Pio ; De Bonis, Maria ; Mancini, Maria Teresa ; Balsamo, Teresa ; Graziano, Paolo ; Centra, Flavia ; Sparaneo, Angelo ; Trombetta, Domenico ; Bonfitto, Antonio ; Scagliusi, Vito ; Larizza, Pietro ; Capoluongo, Ettore Domenico ; Fazio, Vito Michele. / Automated workflow for somatic and Germline next generation sequencing analysis in routine clinical cancer diagnostics. In: Cancers. 2019 ; Vol. 11, No. 11.
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