Comparison of Targeted Next-Generation Sequencing (NGS) and Real-Time PCR in the Detection ofEGFR,KRAS,andBRAFMutations on Formalin-Fixed, Paraffin-Embedded Tumor Material of Non-Small Cell Lung Carcinoma-Superiority of NGS

2013 ◽  
Vol 52 (5) ◽  
pp. 503-511 ◽  
Author(s):  
Katja Tuononen ◽  
Satu Mäki-Nevala ◽  
Virinder Kaur Sarhadi ◽  
Aino Wirtanen ◽  
Mikko Rönty ◽  
...  
Keyword(s):  
Small Cell Lung Carcinoma ◽  
Small Cell Lung ◽  
Cell Lung Carcinoma ◽  
Targeted Next Generation Sequencing ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Tumor Material ◽  
Next Generation Sequencing Ngs ◽  
Generation Sequencing ◽  
Formalin Fixed

Clinical next generation sequencing (NGS) of fine needle aspiration (FNA) biopsies in non-small cell lung (NSCLC) and pancreatic cancers.

2013 ◽  
Vol 31 (15_suppl) ◽  
pp. 11100-11100
Author(s):  
Matthew J. Hawryluk ◽  
Jeffrey S. Ross ◽  
Christine E. Sheehan ◽  
Jie He ◽  
Geneva Young ◽  
...  
Keyword(s):  
Needle Aspiration ◽  
Primary Tumors ◽  
Pancreatic Cancers ◽  
Manual Inspection ◽  
Formalin Fixed Paraffin ◽  
Pancreatic Masses ◽  
Formalin Fixed Paraffin Embedded ◽  
Next Generation Sequencing Ngs ◽  
Generation Sequencing ◽  
Formalin Fixed

11100 Background: FNA is a common diagnostic procedure for the evaluation of pulmonary and pancreatic masses. We sought to determine whether NGS could be performed on these small specimens and to characterize heterogeneity across classes of genomic alterations (GA) in a subset of paired FNA and matched resected primary tumors. Methods: DNA was isolated from formalin fixed paraffin embedded (FFPE) sections of FNA cell blocks using 40µm total sections for NSCLC and 20µm total sections for pancreatic cancers in a CLIA-certified lab (Foundation Medicine). DNA sequencing was performed for 3,320 exons of 182 cancer-related genes and 37 introns of 14 genes frequently rearranged in cancer on indexed, adaptor ligated, hybridization-captured libraries to a median depth of 931x for the NSCLC and 416x for the pancreatic FNAs. Results: Genomic profiles were successfully generated from 19/19 of the NSCLC and 22/23 of the pancreatic FNA cases. We found 97 GA in the 19 NSCLC FNAs (range 2-9, average 5.1 GA per patient). 68% of the patients had GA in TP53 and 21% in KRAS. Only 16% (3/19) patients did not exhibit an actionable alteration. We found 99 GA in the 23 pancreatic cancer FNAs (range 0-12, average 4.3 per patient). 74% of the patients had GA in KRAS. There was 94% concordance of GA found in 4 matched FNA and primary tumor pairs for the pancreatic cases. For the single discordance, manual inspection of the reads of the discordant allele indicated evidence of loss of heterozygosity. Conclusions: NGS can be reliably performed on small FNA samples processed into cell blocks, and the GA discovered significantly correlates with the GA found in matched primary tumors. This study demonstrates the feasibility of NGS in analyzing FNA samples and further broadens the spectrum of commonly encountered specimen types to which this approach can be successfully applied.

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