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Related Experiment Video

Updated: Jun 29, 2025

A Blood-based Test for the Detection of ROS1 and RET Fusion Transcripts from Circulating Ribonucleic Acid Using Digital Polymerase Chain Reaction
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RET Fusion Testing in Patients With NSCLC: The RETING Study.

Esther Conde1,2,3,4, Susana Hernandez1,3, Jose Luis Rodriguez Carrillo5

  • 1Hospital Universitario 12 de Octubre, Madrid, Spain.

JTO Clinical and Research Reports
|March 25, 2024
PubMed
Summary

Accurate detection of RET fusions in non-small cell lung cancer (NSCLC) is crucial for targeted therapy. This study highlights limitations in fluorescence in situ hybridization (FISH) and reverse transcriptase-polymerase chain reaction (RT-PCR) assays, reinforcing the need for upfront next-generation sequencing (NGS).

Keywords:
FISHLung carcinomaNext-generation sequencingRET fusionsRT-PCR

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Area of Science:

  • Oncology
  • Molecular Diagnostics
  • Genomics

Background:

  • RET inhibitors offer significant response rates in non-small cell lung cancer (NSCLC).
  • Accurate identification of RET fusions, a key predictive biomarker, remains challenging.
  • Current guidelines recommend next-generation sequencing (NGS) or alternative methods like fluorescence in situ hybridization (FISH) and reverse transcriptase-polymerase chain reaction (RT-PCR).

Purpose of the Study:

  • To evaluate the real-world performance of various RET fusion detection assays in a multicenter cohort of NSCLC patients.
  • To compare the sensitivity and specificity of NGS, FISH, and RT-PCR for identifying RET fusions.

Main Methods:

  • A cohort of 38 RET fusion-positive NSCLC specimens was analyzed.
  • Specimens were tested using RNA-based NGS (reference standard), two different RET FISH assays, and one RT-PCR assay.
  • Common RET fusion partners and specific fusion events were identified.

Main Results:

  • Next-generation sequencing (NGS) identified a broader range of RET fusions compared to FISH and RT-PCR.
  • Three RET fusions were missed by RT-PCR, and two were missed by FISH, despite being positive by NGS.
  • Histopathological features such as signet ring cells and psammoma bodies were frequently observed in these tumors.

Conclusions:

  • Single-analyte assays like FISH and RT-PCR have limitations and can yield false-negative results for RET fusions.
  • Understanding the performance characteristics of different RET testing methodologies is essential for developing effective diagnostic algorithms.
  • Upfront NGS is reinforced as the preferred method for comprehensive RET fusion detection in NSCLC patients to avoid diagnostic gaps.