| 摘要 |
he invention concerns new prototype protocols for the reverse transcription (RT) and the reaction of the polymerase chain reaction (PCR) for the detection of the molecular marker Ep-Cam and the identification of circulating micrometastatic cancer cells in peripheral blood (and tissues) of patients suffering from cancer of epithelial origin. The detection of circulating micrometastatic cancer cells in the peripheral blood of patients (and in tissues) with cancer of epithilial origin is carriedout with reaction PCR and analysis of the PCR products either through gel agarose electrophoresis or with hybridization in liquid or in membrane with any method, where is used, Neowater, specific primers, where the forward primer will be labelled with biotin in one of the ends and with specific internal probe could be varied. The new invention decreases dramatically the time and the quantity of the reagents, that are sufficient to carry out a detection or test with the use of PCr or/nad RT, while simultaneously is not used for the RT and PCR reaction any buffer and constitutes in the use of total RNA, that could be isolated with any common procedure. A part of the isolated total RNA will be used for the production of complementary DNA (cDNA) using Neowater, a commerciall available reagent. A part of the produced cDNA will be used in the procedure of PCR, eith the use of specific primers, Neowater, prototype thermical protocols and specific internal probe for the hybridization. Thusis detected the Ep-Cam and is carried out the identification of circulating micrometastic cancer cells in peripheral blood or/and in tissues as well as in every other body fluid of patients with cancer of epithelial origin. |
