Process of in vitro detection of humoral rejection risk of transplantation, in a patient having organ transplant, comprises quantitative detection of Cd4 complement components deposited on erythrocytes content in blood sample

摘要

Process of in vitro detection of humoral rejection risk of transplantation, in a patient having an organ transplant, comprises quantitative detection of a Cd4 complement components, deposited on erythrocytes content in a blood sample withdraw from the patient. Independent claims are included for: (1) a process of in vitro quantitative detection of Cd4 complement components deposited on erythrocytes in a blood sample taken from the patient comprising: marking a part using a first molecule associated with a marker having the ability to recognize and bind specifically the Cd4 complement components deposited on erythrocytes of the blood sample, and marking an other parts using a second molecule associated with a marker having the ability to recognize and bind specifically the Cd4 complement components of stabilized erythrocyte compositions or micro particles covered with the Cd4 complement components, where the composition comprises an internal standard for the quantification; introducing the sample for quantifying and the composition of internal standard in a flow cytometry; detecting the signal induced by each of the above markers; and quantifying signal associated with Cd4 complement components deposited on the erythrocytes in the tested sample; (2) a stabilized erythrocyte suspension covered with a known quantity of Cd4 complement components, where the Cd4 complement component is linked by covalent bond directly or indirectly with erythrocytes; and (3) a kit for the detection of the risk in humoral transplant rejection, comprising the compositions constituting an internal standard for quantification Cd4 complement component, where the composition comprising particles/stabilized erythrocyte suspension, covered with Cd4 complement component in the form of Cd4 fragment/a molecule containing Cd4 complement component; a calibration curve for the compositions of internal standard, allowing linking the relative measures of fluorescence intensity with arbitrary units or Cd4 antigenic sites, where the curve was obtained by analysis in flow cytometry, and the background noise optionally may be subtracted to get a corrected value; optionally, a composition of molecules with the ability to recognize and to link to a specific Cd4 complement component, where the molecules being marked e.g. by a fluorochrome.