BACKGROUND: Immunomagnetic enrichment followed by RT-PCR (immunobead RT-PCR) is an efficient methodology to identify disseminated @entity189 cells in the blood and bone marrow. The RT-PCR assays must be both specific for the @entity5 cells and sufficiently sensitive to enable detection of @entity5 cells. We have developed a method to test RT-PCR assays for any @entity5 . This has been investigated using a panel of RT-PCR markers suitable for the detection of @entity0 cells. METHODS: In the assay, a single cell line-derived @entity5 cell is added to 100 peripheral blood mononuclear cells (PBMNCs) after which mRNA is isolated and reverse transcribed for RT-PCR analysis. PBMNCs without added @entity5 cells are used as specificity controls. The previously studied markers @entity470 ( @entity470 ), @entity36637 ( @entity36637 ), @entity5960 ( @entity5960 / @entity5960 ), @entity2355 ( @entity2355 ), @entity10580 ( @entity10580 ) were tested. Two new epithelial-specific markers @entity18437 and @entity25335 were also tested. RESULTS: @entity2355 was unsuitable as strong amplification was detected in 100 cell PBMNC controls. Expression of @entity18437 , @entity25335 , @entity5960 , @entity470 , @entity10580 and @entity36637 was found to be both specific for the @entity5 cell, as demonstrated by the absence of a signal in most 100 cell PBMNC controls, and sensitive enough to detect a @entity5 cell in 100 PBMNCs using a single round of RT-PCR. CONCLUSIONS: @entity18437 , @entity25335 , @entity5960 , @entity470 , @entity10580 and @entity36637 are appropriate RT-PCR markers for use in a marker panel to detect disseminated @entity0 cells after immunomagnetic enrichment.

Optimisation of the RT-PCR detection of immunomagnetically enriched XXXX cells.

@entity5960 :: ('4072', 'Gene') :: ['TACSTD1', 'epithelial cell adhesion molecule', 'EpCAM'] @entity0 :: ('MESH:D001943', 'Disease') :: ['breast cancer'] @entity5 :: ('MESH:D009369', 'Disease') :: ['tumor', 'single tumor', 'cancer'] @entity36637 :: ('4250', 'Gene') :: ['MGB1', 'mammaglobin 1'] @entity470 :: ('1956', 'Gene') :: ['EGFR', 'epidermal growth factor receptor'] @entity18437 :: ('1999', 'Gene') :: ['ELF3'] @entity189 :: ('MESH:D002277', 'Disease') :: ['carcinoma'] @entity25335 :: ('2050', 'Gene') :: ['EphB4'] @entity10580 :: ('1048', 'Gene') :: ['CEA', 'carcinoembryonic antigen'] @entity2355 :: ('4582', 'Gene') :: ['MUC1', 'mucin 1']
@entity189 :: (MESH:D002277,Disease) :: ['carcinoma']
