Analysis of pre-analytic factors affecting the success of clinical next-generation sequencing of solid organ malignancies

Hui Chen, Rajyalakshmi Luthra, Rashmi S. Goswami, Rajesh R. Singh, Sinchita Roy-Chowdhuri

Research output: Contribution to journalReview articlepeer-review

94 Scopus citations

Abstract

Application of next-generation sequencing (NGS) technology to routine clinical practice has enabled characterization of personalized cancer genomes to identify patients likely to have a response to targeted therapy. The proper selection of tumor sample for downstream NGS based mutational analysis is critical to generate accurate results and to guide therapeutic intervention. However, multiple pre-analytic factors come into play in determining the success of NGS testing. In this review, we discuss pre-analytic requirements for AmpliSeq PCR-based sequencing using Ion Torrent Personal Genome Machine (PGM) (Life Technologies), a NGS sequencing platform that is often used by clinical laboratories for sequencing solid tumors because of its low input DNA requirement from formalin fixed and paraffin embedded tissue. The success of NGS mutational analysis is affected not only by the input DNA quantity but also by several other factors, including the specimen type, the DNA quality, and the tumor cellularity. Here, we review tissue requirements for solid tumor NGS based mutational analysis, including procedure types, tissue types, tumor volume and fraction, decalcification, and treatment effects.

Original languageEnglish (US)
Pages (from-to)1699-1715
Number of pages17
JournalCancers
Volume7
Issue number3
DOIs
StatePublished - Aug 28 2015

Keywords

  • Next-generation sequencing
  • Pre-analytic factors
  • Targeted hotspot mutation analysis
  • Tissue qualification

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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