The present invention concerns: - devices in the shape of microplates or microstrips having wells with extended length apt to receive 3 or 4 or 6 immunosorbent elements protruding from a rod at the same modular distance of wells arranged in standard 8-wells microstrips or in 12-wells microstrips or in standard 96-wells microplates; - a method that uses these devices making two different shaped solid phases compete with each other; a solid phase is constituted by the extended wells, each of which immobilize a different type of cell or molecule in the examined sample; the other solid phase is constituted by immunosorbent elements protruding from a rod, each of which has been previously coated with one of the same markers to be detected in the sample; then, these immunosorbent elements are dipped into each extended well in groups of 3 or 4 or 6, after ligands for markers to be detected have been added in liquid phase in the extended wells; these ligands bind to the immunosorbent elements in a inversely proportional amount to the quantity of the markers of each type of cell or molecule immobilized on the proper extended well that competes for the binding; these ligands are simultaneously quantifiable by an immunoenzymatic assay carried out by dipping the rod with the immunosorbent elements into a tube containing the conjugate and, then, by dipping each single element into the wells of standard microplates or microstrips containing chromogenic substrate, where the spectrophotometric reading is finally performed; - kits suited for carrying out the above said method, using the above said devices with extended wells in order to obtain the simultaneously quantification of different cellular or molecular markers and the simultaneous detection of different types of cells or molecules exhibiting them, including immunoassay kits for a rapid tuberculosis diagnosis and paratuberculosis diagnosis at early stage, based on the quantification of different infection markers of different cooperating and cytolytic lymphocyte populations sensitized against different antigens or mycobacterial haptens.
EP2699908A1 Device, method and kit for the detection of different markers in different cellular or molecular types and their quantification
MAZZEO, Alessandra
2014-01-01
Abstract
The present invention concerns: - devices in the shape of microplates or microstrips having wells with extended length apt to receive 3 or 4 or 6 immunosorbent elements protruding from a rod at the same modular distance of wells arranged in standard 8-wells microstrips or in 12-wells microstrips or in standard 96-wells microplates; - a method that uses these devices making two different shaped solid phases compete with each other; a solid phase is constituted by the extended wells, each of which immobilize a different type of cell or molecule in the examined sample; the other solid phase is constituted by immunosorbent elements protruding from a rod, each of which has been previously coated with one of the same markers to be detected in the sample; then, these immunosorbent elements are dipped into each extended well in groups of 3 or 4 or 6, after ligands for markers to be detected have been added in liquid phase in the extended wells; these ligands bind to the immunosorbent elements in a inversely proportional amount to the quantity of the markers of each type of cell or molecule immobilized on the proper extended well that competes for the binding; these ligands are simultaneously quantifiable by an immunoenzymatic assay carried out by dipping the rod with the immunosorbent elements into a tube containing the conjugate and, then, by dipping each single element into the wells of standard microplates or microstrips containing chromogenic substrate, where the spectrophotometric reading is finally performed; - kits suited for carrying out the above said method, using the above said devices with extended wells in order to obtain the simultaneously quantification of different cellular or molecular markers and the simultaneous detection of different types of cells or molecules exhibiting them, including immunoassay kits for a rapid tuberculosis diagnosis and paratuberculosis diagnosis at early stage, based on the quantification of different infection markers of different cooperating and cytolytic lymphocyte populations sensitized against different antigens or mycobacterial haptens.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
