RT-PCR Assays

These unique RT-PCR (reverse transcriptase polymerase chain reaction) Beacon Assays for research can be used to identify the presence of occult breast epithelial cells in lymph node, bone marrow, or peripheral blood samples. All four quantitative homogeneous assays utilize one-step RT-PCR technology.  Vysis^® selected nucleic acid amplification as the technology of choice for this application because there is:

• Research and clinical diagnostic acceptance
• Wide reagent and instrumentation availability
• High-throughput and multiplexing potential
• Value as a sensitive and accurate methodology

CK19 RT-PCR Beacon Assay
The Cytokeratin 19 (CK19) gene is a good general marker of epithelial cells.Combined with our proprietary breast-specific markers, CK19 offers the opportunity to identify an epithelial cell along with its tissue of origin. The CK19 RT-PCR Beacon Assay was specifically designed to avoid amplification of any of the identified pseudogenes. In addition, this assay may also be used for the detection of other epithelial cancers types.

B2M RT-PCR Beacon Assay
The Beta-2 Microglobulin gene (B2M) has been identified as a good "housekeeping" gene for assessing RNA integrity. In addition to assessing the quality of a sample, the B2M RT-PCR Beacon Assay is useful for the quantification of peripheral blood leukocytes (PBLs) in an enriched blood sample.

BS106 RT-PCR Beacon Assay
The BS106 gene encodes for a secreted,mucin-like protein specifically expressed by breast epithelial cells.1 It is also known in the literature as SBEM. This novel, proprietary marker can be combined with other markers of breast cells to produce a panel for the detection of rare cell events in this disease.

BU101 RT-PCR Beacon Assay
The BU101 gene is specifically expressed by breast epithelial cells. similar to mammaglobin,which are both members of the secretogobulin family of proteins. The function(s) of BU101 is not known. This novel, proprietary marker can be combined
with other markers of breast cells to produce a panel for the detection of rare cell events in this disease.
References
1.Colpitts TL, Billing-Medel P,Granados EN, et al. Tumor Biol. 23 (2002): 263-278.
2.Colpitts TL, Billing-Medel P, Friedman P, et al. Biochemistry, 40 (2001): 11048-11059.

Product	Description	Specifications
CK19 RT-PCR Beacon Assay	Sequence-specific primers and probe for quantitative, real-time RT-PCR detection of Epithelial Marker CK19 mRNA	1 vial containing CK19 forward primer, CK19 reverse primer, and CK 19 FAM-labeled Beacon probe in TE buffer
B2M RT-PCR Beacon Assay	Sequence-specific primers and probe for quantitative, real-time RT-PCR detection of Housekeeping Gene B2M mRNA	1 vial containing B2M forward primer, B2M reverse primer, and B2M VIC-labeled Beacon probe in TE buffer
BS106 RT-PCR Beacon Assay	Sequence-specific primers and probe for quantitative, real-time RT-PCR detection of Breast Marker BS106 mRNA	1 vial containing BS106 forward primer, BS106 reverse primer, and BS106 FAM-labeled Beacon probe in TE buffer
BU101 RT-PCR Beacon Assay	Sequence-specific primers and probe for quantitative, real-time RT-PCR detection of Breast Marker BU101 mRNA	1 vial containing BU101 forward primer, BU101 reverse primer, and BU101 FAM-labeled Beacon probe in TE buffer