title: Application of DNA-based forensic analysis for the detection of homologous transfusion of whole blood and of red blood cell concentrates in doping control.
abstract: In this work we present the application of a method for the identification of homologous blood transfusions using forensic genetic techniques based on DNA typing. Ex vivo mixtures of human blood samples - either whole blood or red blood cell concentrates - simulating homologous blood transfusions at different percentages of the donor were typed for a panel of 16 highly variable DNA short tandem repeats (STR). Tested samples included also mixtures, which gave false-negative results if assayed by the reference flow cytofluorimetric method, which is based on the recognition of target antigens located on the membrane of the red blood cell. The recognition of triplets and quadruplets at various loci gave information of the presence of cells belonging to different individuals, as it is the case for homologous blood transfusions. Specificity and sensitivity of the method were assessed in the validation study. The method proved to be unequivocally specific since it was able to recognize all single profiles of each individual, clearly discriminating them from mixtures. Sensitivity resulted as a consequence of the percentage of the donor aliquot in the total volume of the mixture. Although the source of DNA in a blood sample is represented only by nucleated white blood cells, the same procedure resulted effective also in detecting mixtures of red blood cell concentrates (RBCC) from leukodepletion procedure: DNA of the donor from the residual white blood cells resulted still detectable, even if with an expected loss of sensitivity. The proposed approach may contribute to reduce the risk of false-negative results, which may occur using the reference cytofluorimetric method.
authors.affiliation: Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti 1, 00197 Rome, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Stampella
authors.affiliation: Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti 1, 00197 Rome, Italy.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Di Marco
authors.affiliation: Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti 1, 00197 Rome, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Pirri
authors.affiliation: Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti 1, 00197 Rome, Italy.
authors.forename: Xavier
authors.initials: X
authors.lastname: de la Torre
authors.affiliation: Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti 1, 00197 Rome, Italy; Dipartimento di Medicina Sperimentale, "Sapienza" Università di Roma, Viale Regina Elena 324, 00161 Rome, Italy. Electronic address: francesco.botre@uniroma1.it.
authors.forename: Francesco
authors.initials: F
authors.lastname: Botrè
authors.affiliation: Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti 1, 00197 Rome, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Donati
mesh_terms: D001803:Blood Transfusion; D004247:DNA / Q000032:analysis*; D004300:Doping in Sports; D053803:Forensic Genetics; D006801:Humans; D012680:Sensitivity and Specificity
chemical_list: D004247:DNA
keywords: Blood doping; DNA typing; Doping analysis; Homologous blood transfusion; Red blood cells concentrates; Short tandem repeats