Potential pitfalls in multiplex PCR-based next-generation sequencing: a case-based report

J Clin Pathol. 2023 Jan;76(1):59-63. doi: 10.1136/jclinpath-2021-208105. Epub 2022 Feb 10.

Abstract

Amplicon-based next-generation sequencing (NGS) assays employ highly sensitive, rapid, and cost-effective methods to detect clinically actionable mutations for the diagnosis, prognosis, and treatment of patients with cancer. However, recognition of certain limitations inherent to amplicon-based NGS assays is crucial for the correct interpretation and reporting of variants in the clinical setting. In this report, we illustrate three different potential pitfalls related to amplicon-based NGS assays based on our institutional experience and highlight how the risk of such events can be minimised.

Keywords: genes; genetics; medical oncology; molecular; neoplasm; pathology.

MeSH terms

  • High-Throughput Nucleotide Sequencing
  • Humans
  • Multiplex Polymerase Chain Reaction*
  • Mutation
  • Neoplasms* / genetics
  • Prognosis