scholarly journals Targeted Resequencing RevealsALKFusions in Non-Small Cell Lung Carcinomas Detected by FISH, Immunohistochemistry, and Real-Time RT-PCR: A Comparison of Four Methods

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Katja Tuononen ◽  
Virinder Kaur Sarhadi ◽  
Aino Wirtanen ◽  
Mikko Rönty ◽  
Kaisa Salmenkivi ◽  
...  

Anaplastic lymphoma receptor tyrosine kinase (ALK) gene rearrangements occur in a subgroup of non-small cell lung carcinomas (NSCLCs). The identification of these rearrangements is important for guiding treatment decisions. The aim of our study was to screenALKgene fusions in NSCLCs and to compare the results detected by targeted resequencing with results detected by commonly used methods, including fluorescence in situ hybridization (FISH), immunohistochemistry (IHC), and real-time reverse transcription-PCR (RT-PCR). Furthermore, we aimed to ascertain the potential of targeted resequencing in detection ofALK-rearranged lung carcinomas. We assessedALKfusion status for 95 formalin-fixed paraffin-embedded tumor tissue specimens from 87 patients with NSCLC by FISH and real-time RT-PCR, for 57 specimens from 56 patients by targeted resequencing, and for 14 specimens from 14 patients by IHC. All methods were performed successfully on formalin-fixed paraffin-embedded tumor tissue material. We detectedALKfusion in 5.7% (5 out of 87) of patients examined. The results obtained from resequencing correlated significantly with those from FISH, real-time RT-PCR, and IHC. Targeted resequencing proved to be a promising method forALKgene fusion detection in NSCLC. Means to reduce the material and turnaround time required for analysis are, however, needed.

Author(s):  
Janne Lehtiö ◽  
Taner Arslan ◽  
Ioannis Siavelis ◽  
Yanbo Pan ◽  
Fabio Socciarelli ◽  
...  

Abstract The associated publication reports proteogenomic analysis of non-small cell lung cancer, where we identified molecular subtypes with distinct immune evasion mechanisms and therapeutic targets, and validated our classification method in separate clinical cohorts. This protocol describes histological, tertiary lymphoid structure (TLS), and immunohistochemical evaluation of clinical samples. Specifically, immunohistochemistry was performed for PD-L1, CD3, and CD8 on tumor microarrays (TMAs) derived from formalin-fixed paraffin embedded (FFPE) samples.


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