title: Multiplexed tandem polymerase chain reaction identifies strong expression of oestrogen receptor and Her-2 from single, formalin-fixed, paraffin-embedded breast cancer sections.
abstract: OBJECTIVE
  To establish the suitability of multiplex tandem polymerase chain reaction (MT-PCR) for rapid identification of oestrogen receptor (ER) and Her-2 status using a single, formalin-fixed, paraffin-embedded (FFPE) breast tumour section.
  
  
  METHODS
  Tissue sections from 29 breast tumours were analysed by immunohistochemistry (IHC) and fluorescence in situ hybridisation (FISH). RNA extracted from 10 mum FFPE breast tumour sections from 24 of 29 tumours (14 ER positive and 5 Her-2 positive) was analysed by MT-PCR. After establishing a correlation between IHC and/or FISH and MT-PCR results, the ER/Her-2 status of a further 32 randomly selected, archival breast tumour specimens was established by MT-PCR in a blinded fashion, and compared to IHC/FISH results.
  
  
  RESULTS
  MT-PCR levels of ER and Her-2 showed good concordance with IHC and FISH results. Furthermore, among the ER positive tumours, MT-PCR provided a quantitative score with a high dynamic range. Threshold values obtained from this data set applied to 32 archival tumour specimens showed that tumours strongly positive for ER and/or Her-2 expression were easily identified by MT-PCR.
  
  
  CONCLUSIONS
  MT-PCR can provide rapid, sensitive and cost-effective analysis of FFPE material and may prove useful as triage to identify patients suited to endocrine or trastuzumab (Herceptin) treatment.
authors.affiliation: University of Melbourne, Department of Surgery, Australia. rik@medstv.unimelb.edu.au
authors.forename: Erik W
authors.initials: EW
authors.lastname: Thompson
authors.forename: Kristina
authors.initials: K
authors.lastname: Warton
authors.forename: Tony
authors.initials: T
authors.lastname: Blick
authors.forename: Razan
authors.initials: R
authors.lastname: Wafai
authors.forename: Prue
authors.initials: P
authors.lastname: Hill
authors.forename: Keith
authors.initials: K
authors.lastname: Stanley
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D001943:Breast Neoplasms / Q000191:economics / Q000235:genetics / Q000378:metabolism*; D016172:DNA Fingerprinting; D004273:DNA, Neoplasm / Q000032:analysis; D005260:Female; D005557:Formaldehyde / Q000737:chemistry; D017048:Health Care Costs; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000191:economics / Q000379:methods; D009363:Neoplasm Proteins / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D016612:Paraffin Embedding / Q000379:methods; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism*; D011960:Receptors, Estrogen / Q000378:metabolism*; D015203:Reproducibility of Results; D020080:Tandem Repeat Sequences / Q000235:genetics; D016707:Tissue Fixation
chemical_list: D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; D005557:Formaldehyde; D009363:Neoplasm Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D011960:Receptors, Estrogen; C508053:ERBB2 protein, human