Minimal residual breast cancer detected using CK19 and EGF-R primers in RT-PCR.

2004 JUN 3 - (NewsRx.com & NewsRx.net) -- Minimal residual breast cancer was detected using cytokeratin 19 and epidermal growth factor receptor primers in an RT-PCR assay.

"Breast cancer cells pose a difficult target for detection due to their lack of tumor specific markers. Small numbers of metastatic cells circulate through the blood and reach target organs, but current methods are presently not sensitive enough to allow for early detection. Our goal is to develop a method for specific and sensitive detection for breast cancer tumor cells by using cell enrichment coupled to RT-PCR with selective primer pairs," wrote scientists in the International Journal of Oncology.

"Fifty million normal donor mononuclear cells (NDMNC) were spiked with serial dilutions of a breast cancer cell line, positively selected for surface expression of the epithelial cell adhesion molecule (EPCAM) with anti-Ber-EP4 mAb. Total RNA was isolated and the cDNA was transcribed and measured by quantitative PCR for cytokeratin 19 (CK19), epidermal growth factor receptor (EGF-R), and B-2-microglobulin (beta2M) transcripts using specific hybridization probes with the LightCycler System," V.S. Fellowes and colleagues reported.

"A sensitivity of 1 tumor cell in 5 million NDMNC was consistently achieved with a metastatic breast cancer cell line with target primer sets for CK19 and EGF-R. Less, but appreciable sensitivity was achieved by spiking NDMNC with other breast ...