A method for detecting circulating tumor cells, and apoptotic bodies microembolisms IN BLOOD OF PATIENTS WITH LUNG CANCER MAN

摘要

1. A method for the detection of circulating tumor cells, apoptotic bodies and microemboli in the blood of human patients with lung cancer, comprising centrifuging a sample of peripheral blood of patients with lung cancer, the separation of blood plasma, the concentrated cell pellet incubated with a hypotonic ammonium chloride solution to the destruction of red blood cells, the remaining cell resuspension in phosphate buffer, microscopic examination of the sample on slides, characterized in that before the step of resuspension remaining after fracture I cell erythrocytes were pelleted by centrifugation and by addition of a hypotonic solution of sodium chloride partly destroy leukocytes, more sample is centrifuged and removed the supernatant, the pellet containing unbroken cells, resuspended in phosphate buffer containing Cai Mg, to restore the volume of the remaining cells, and the cells were concentrated to reduce total sample volume, the resulting cell suspension was incubated with the masking DNA then added aptamers specific for lung cancer, a fluorescent labeled IU Coy, the samples were examined by microscopy techniques that allow fluorescent metki.2 register. A method according to claim 1, characterized in that the aptamers specific for human lung tumor tissues, obtained during the breeding, having established the nucleotide sequence reported as SEQ ID NO 1 - SEQ ID NO 34.3. A method according to claim 1, characterized in that after the step of incubating the cell suspension with masking DNA aptamers added simultaneously with monoclonal antibodies labeled with a fluorescent label, spec