Doing more with less: fluorescence in situ hybridization and gene sequencing assays can be reliably performed on archival stained tumor tissue sections
| Autor: | Ugo Pastorino, Alessandra Fabbri, Elena Tamborini, Enrica Bovio, Marina Chiara Garassino, Adele Busico, Benedetta Picciani, Barbara Valeri, Giulio Settanni, Angelica Sonzogni, Federica Perrone, Giuseppe Pelosi, Filippo de Braud, Maria Adele Testi |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Adult Male Pathology medicine.medical_specialty Adolescent DNA Mutational Analysis H&E stain Biology medicine.disease_cause DNA sequencing Pathology and Forensic Medicine 03 medical and health sciences Young Adult 0302 clinical medicine Neoplasms medicine Biomarkers Tumor Humans Molecular Biology In Situ Hybridization Fluorescence Aged Aged 80 and over medicine.diagnostic_test Staining and Labeling High-Throughput Nucleotide Sequencing Cell Biology General Medicine Middle Aged Tumor tissue Molecular biology Immunohistochemistry 030104 developmental biology 030220 oncology & carcinogenesis Fish Alkaline phosphatase Feasibility Studies Female KRAS Fluorescence in situ hybridization |
| Zdroj: | Virchows Archiv : an international journal of pathology. 468(4) |
| ISSN: | 1432-2307 |
| Popis: | Little is known about molecular testing on tumor tissue retrieved from stained sections, for which there may be a clinical need. We retrospectively analyzed 112 sections from 56 tumor patients using either fluorescence in situ hybridization (FISH) with different probes (19 sections from 17 patients) or Sanger or targeted next generation sequencing for detection of BRAF, EGFR, KRAS, C-KIT, and TP53 mutations (93 sections from 39 patients). Tumor tissue sections had been stained by hematoxylin and eosin (H&E) (42 sections) or by immunohistochemistry for cytoplasmic or nuclear/nuclear-cytoplasmic markers (70 sections) with a peroxidase (P-IHC, with 3,3'-diaminobenzidine as chromogen) or alkaline phosphatase label (AP-IHC, with Warp Red™ as chromogen). For FISH analysis, the concordance rate between the original diagnosis and that obtained on H&E- or P-IHC-stained tissue sections (AP-IHC was not on record for this set of patients) was 95% (18 out of 19 tumor sections). Only one tumor sample, diffusely positive for MLH1, did not yield any nuclear hybridization signal. For sequencing analysis, the concordance rate was 100% on negative P-IHC and positive AP-IHC-stained sections, regardless of the subcellular localization of the reaction product. Mutations were detected in only 52% of cases expressing nuclear/nuclear-cytoplasmic markers, regardless of the sequencing technology used (p = 0.0002). In conclusion, stained sections may be a valuable resource for FISH or sequencing analysis, but on cases expressing nuclear markers sequencing results need to be interpreted cautiously. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink