title: Utility of sarcoma-specific fusion gene analysis in paraffin-embedded material for routine diagnosis at a specialist centre.
abstract: OBJECTIVE
  Diagnosis of soft tissue sarcomas can be difficult. It can be aided by detection of specific genetic aberrations in many cases. This study assessed the utility of a molecular genetics/cytogenetics service as part of the routine diagnostic service at the Royal Marsden Hospital.
  
  
  METHODS
  A retrospective audit was performed over a 15-month period to evaluate the diagnostic usefulness for soft tissue sarcomas with translocations of fluorescence in situ hybridisation (FISH) and reverse-transcriptase PCR (RT-PCR) in paraffin-embedded (PE) material. Results were compared with histology, and evaluated.
  
  
  RESULTS
  Molecular investigations were performed on PE material in 158 samples (total 194 RT-PCR and 174 FISH tests), of which 85 were referral cases. Synovial sarcoma, Ewing sarcoma and low-grade fibromyxoid sarcoma were the most commonly tested tumours. Myxoid liposarcoma showed the best histological and molecular concordance, and alveolar rhabdomyosarcoma showed the best agreement between methods. FISH had a higher sensitivity for detecting tumours (73%, compared with 59% for RT-PCR) with a better success rate than RT-PCR, although the latter was specific in identifying the partner gene for each fusion. In particular, referral blocks in which methods of tissue fixation and processing were not certain resulted in higher RT-PCR failure rates.
  
  
  CONCLUSIONS
  FISH and RT-PCR on PE tissue are practical and effective ancillary tools in the diagnosis of soft tissue sarcomas. They are useful in confirming doubtful histological diagnoses and excluding malignant diagnoses. PCR is less sensitive than FISH, and the use of both techniques is optimal for maximising the detection rate of translocation-positive sarcomas.
authors.affiliation: Department of Histopathology, Royal Marsden Hospital, London, UK. khin.thway@rmh.nhs.uk
authors.forename: Khin
authors.initials: K
authors.lastname: Thway
authors.forename: Sasha
authors.initials: S
authors.lastname: Rockcliffe
authors.forename: David
authors.initials: D
authors.lastname: Gonzalez
authors.forename: John
authors.initials: J
authors.lastname: Swansbury
authors.forename: Toon
authors.initials: T
authors.lastname: Min
authors.forename: Lisa
authors.initials: L
authors.lastname: Thompson
authors.forename: Cyril
authors.initials: C
authors.lastname: Fisher
mesh_terms: D003937:Diagnosis, Differential; D004812:Epidemiologic Methods; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D050596:Oncogene Fusion; D016612:Paraffin Embedding; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012509:Sarcoma / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D012983:Soft Tissue Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D014178:Translocation, Genetic