title: A seminested RT-PCR assay for HER2/neu: initial validation of a new method for the detection of disseminated breast cancer cells.
abstract: BACKGROUND
  Initial validation of a seminested reverse transcription-polymerase chain reaction (RT-PCR) assay for HER2/neu for use in detecting circulating tumor cells in the peripheral blood or bone marrow of breast cancer patients is described. RT-PCR assays for other epithelial markers, including the cytokeratins and carcinoembryonic antigen frequently lack specificity, sensitivity, or both. Thus, there is a need for an assay that is both sensitive and specific to be used to monitor breast cancer patients for micrometastatic or minimal residual disease.
  
  
  RESULTS
  Assay conditions were optimized using the MCF7 breast cancer cell line and the Raji B-cell lymphoma cell line. The assay can detect as little as 3 mg of MCF7 RNA within a background of 3 mg of Raji RNA. The assay was positive in 12 of 12 breast tumors. None of the 33 peripheral blood or stem cell samples form patients without evidence of breast cancer were positive. Peripheral blood from 17 breast cancer patients was collected immediately before surgery and evaluated. The assay was positive in five of six patients with Stage II, four of eight patients with Stage I and one of three patients with Stage 0 disease.
  
  
  CONCLUSIONS
  The seminested HER2/neu RT-PCR assay compares favorably with RT-PCR assays for other epithelial cell markers in terms of both sensitivity and specificity as a method to detect disseminated breast cancer cells. In breast cancer patients, the higher the disease stage, the more frequently was the assay positive.
authors.affiliation: Division of Medical Genetics, Department of Medicine, UCSD Cancer Center, University of California, San Diego, La Jolla 92093-0639, USA.
authors.forename: L
authors.initials: L
authors.lastname: Wasserman
authors.forename: A
authors.initials: A
authors.lastname: Dreilinger
authors.forename: D
authors.initials: D
authors.lastname: Easter
authors.forename: A
authors.initials: A
authors.lastname: Wallace
mesh_terms: D001943:Breast Neoplasms / Q000097:blood* / Q000235:genetics / Q000473:pathology; D002051:Burkitt Lymphoma / Q000737:chemistry / Q000473:pathology; D004273:DNA, Neoplasm / Q000097:blood / Q000235:genetics*; D005069:Evaluation Studies as Topic; D005260:Female; D018734:Genes, erbB-2; D006801:Humans; D009362:Neoplasm Metastasis; D009367:Neoplasm Staging; D018365:Neoplasm, Residual; D009360:Neoplastic Cells, Circulating; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured / Q000737:chemistry
chemical_list: D004273:DNA, Neoplasm