Related Experiment Video
Updated: Mar 3, 2026
Author Spotlight: Advancements in Molecular Biomarker Testing for Non-Squamous Non-Small Cell Lung Cancer
Published on: September 8, 2023
A highly specific and sensitive massive parallel sequencer-based test for somatic mutations in non-small cell lung
Yoshiaki Inoue1,2, Jun Shiihara3, Hitoshi Miyazawa4
1Graduate School, Saitama Medical University, Moroyama, Saitama, Japan.
Abstract:
Molecular targeting therapy for non-small cell lung cancer (NSCLC) has clarified the importance of mutation testing when selecting treatment regimens. As a result, multiple-gene mutation tests are urgently needed. We developed a next-generation sequencer (NGS)-based, multi-gene test named the MINtS for investigating driver mutations in both cytological specimens and snap-frozen tissue samples. The MINtS was used to investigate the EGFR, KRAS, BRAF genes from DNA, and the ERBB2, and the ALK, ROS1, and RET fusion genes from RNA. We focused on high specificity and sensitivity (≥0.99) and even included samples with a cancer cell content of 1%. The MINtS enables testing of more than 100 samples in a single run, making it possible to process a large number of samples submitted to a central laboratory, and reducing the cost for a single sample. We investigated 96 cytological samples and 190 surgically resected tissues, both of which are isolated in daily clinical practice. With the cytological samples, we compared the results for the EGFR mutation between the MINtS and the PNA-LNA PCR clamp test, and their results were 99% consistent. In the snap-frozen tissue samples, 188/190 (99%) samples were successfully analyzed for all genes investigated using both DNA and RNA. Then, we used 200 cytological samples that were serially isolated in clinical practice to assess RNA quality. Using our procedure, 196 samples (98%) provided high-quality RNA suitable for analysis with the MINtS. We concluded that the MINtS test system is feasible for analyzing "druggable" genes using cytological samples and snap-frozen tissue samples. The MINtS will fill a needs for patients for whom only cytological specimens are available for genetic testing.
Insights
A new multi-gene test, MINtS, accurately detects driver mutations in non-small cell lung cancer using both tissue and cytology samples. This next-generation sequencing test is crucial for personalized molecular targeting therapy selection.
Area of Science:
- Oncology
- Genetics
- Molecular Biology
Background:
- Molecular targeting therapy for non-small cell lung cancer (NSCLC) necessitates comprehensive mutation testing for effective treatment selection.
- Current diagnostic methods often require substantial tissue, limiting genetic analysis in patients with limited samples.
Purpose of the Study:
- To develop and validate a next-generation sequencer (NGS)-based, multi-gene test (MINtS) for investigating driver mutations in NSCLC.
- To assess the feasibility and accuracy of the MINtS test using both cytological specimens and snap-frozen tissue samples.
- To address the urgent need for efficient and cost-effective mutation testing in routine clinical practice.
Main Methods:
- Developed the MINtS, an NGS-based assay targeting EGFR, KRAS, BRAF (DNA), and ERBB2, ALK, ROS1, RET (RNA) fusion genes.
- Validated the assay on 96 cytological samples and 190 snap-frozen tissue samples, including those with as low as 1% cancer cell content.
- Assessed RNA quality from 200 cytological samples to ensure suitability for MINtS analysis.
Main Results:
- The MINtS demonstrated high specificity and sensitivity (≥0.99) across tested genes.
- 99% consistency was observed between MINtS and PNA-LNA PCR clamp tests for EGFR mutations in cytological samples.
- Successful analysis of 99% of tissue samples for all investigated genes (DNA and RNA), with 98% of cytological samples yielding high-quality RNA.
Conclusions:
- The MINtS test system is a feasible and accurate method for analyzing "druggable" genes in NSCLC.
- This assay effectively utilizes both cytological and tissue samples, expanding genetic testing options for patients.
- MINtS addresses a critical need for comprehensive mutation profiling, particularly when only cytological specimens are available.
