RRC ID 68853
Author Inoue Y, Shiihara J, Miyazawa H, Ohta H, Higo M, Nagai Y, Kobayashi K, Saijo Y, Tsuchida M, Nakayama M, Hagiwara K.
Title A highly specific and sensitive massive parallel sequencer-based test for somatic mutations in non-small cell lung cancer.
Journal PLoS One
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.
Volume 12(4)
Pages e0176525
Published 2017-1-1
DOI 10.1371/journal.pone.0176525
PII PONE-D-16-44236
PMID 28448556
PMC PMC5407820
MeSH Carcinoma, Non-Small-Cell Lung / genetics* Carcinoma, Non-Small-Cell Lung / pathology Cryopreservation DNA Mutational Analysis / methods* Humans Lung Neoplasms / genetics* Lung Neoplasms / pathology Mutation* Research Design Software
IF 2.74
Human and Animal Cells PC-9(RCB4455)