Real-time RT-PCR correlates with immunocytochemistry for the detection of disseminated epithelial cells in bone marrow aspirates of patients with breast cancer

摘要: Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) is a technique with the potential of improving quantification disseminated epithelial cells (DEC) in haematological tissues due to its exquisite sensitivity. This sensitivity may lead false positivity. Immunocytochemistry (ICC) regarded as standard methodology diagnose DEC. In this study, detection ICC was compared quantitative real-time RT-PCR for CK-19 and mammaglobin (hMAM) mRNA bone marrow (BM) patients metastatic breast cancer (MBC). Bone aspirated from 14 control 29 MBC. Mononuclear (MNC) were isolated. Immunostaining carried out Epimet kit. Quantitative PCR performed on ABI Prism 7700. The hMAM quantities normalised against beta-Actin calculated relative calibrator sample (relative gene expression). All controls negative by expression measured RT-PCR, whereas median RGE value 0.57. For MBC patients, 0 10 25 (40%) tested positive. Median 2.9 20 (80%) With ICC, 1 stained cell per sample, 15 24 (62%) samples A correlation observed between (r=0.7; P=0.0003), (r=0.6; P=0.003). (r=0.9; P<0.0001) showed strongest correlation. Reverse resulted higher number positive BM than ICC. Since an excellent probably more sensitive advantage being less observer dependent thus also easy automate, we consider our method validated DEC patients.