Immunobead multiplex RT-PCR detection of carcinoembryonic genes expressing cells in the blood of colorectal cancer patients.

摘要: Circulating cell detection using reverse transcriptase-polymerase chain reaction (RT-PCR) techniques has been studied as a new prognostic factor in colorectal cancer patients. With the view of enhancing sensitivity, we developed multiplex RT-PCR assay for circulating based on expression carcinoembryonic antigen-related adhesion molecule 5 (CEACAM5; formerly CEA) and CEACAM7 (formerly CGM2). Between November 2002 December 2003, 45 stage III-IV, 39 I-II patients, 32 non-colorectal patients 41 healthy individuals were included. Positive selection HEA-125 immunobeads was applied to blood samples before mRNA extraction, cDNA synthesis CEACAM5/CEACAM7 assay. For both CEACAM5 CEACAM7, limit found be low 1 expressing 10(6) nucleated cells. The negative test positive 53/84 (63%) and/or whereas 32/84 (38%) markers. Colorectal one two markers 80% cases (36/45) III-IV (CEACAM5 73%, 51%) 44% (17/39) This with proved more sensitive than use single marker detecting tumour discrepant may label cells that have different levels differentiation subsequent aggressive behaviour.