Donati Francesco, de la Torre Xavier, Pagliarosi Sarajane, Pirri Daniela, Prevete Giuliana, Botrè Francesco
Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Rome, Italy.
REDs-Research and Expertise in AntiDoping Sciences, Synathlon, Quartier Centre, ISSUL-Institute of Sport Sciences, University of Lausanne, Lausanne, Switzerland.
Front Sports Act Living. 2022 Feb 10;4:808449. doi: 10.3389/fspor.2022.808449. eCollection 2022.
This article presents the results of a study aimed to give new suggestions and strategies for improving the implementation of the flow cytofluorimetry-based method for the detection of homologous blood transfusions in doping control. The method is based on the recognition of the phenotypic mismatch between minority blood group antigens possessed by the donor and the recipient. Two strategies have been followed to reduce the risk of false-negative results: (i) the monitoring of a broader range of erythrocytes surface antigens; and (ii) the application of different surface erythrocyte staining protocols, tailored on the different antigens and the type of antigenic mismatch that had to be detected (whether it is the donor or the recipient who expresses or not the antigen to be detected). Special attention has also been focused on the time factor, to avoid prolonged sample storage, since hemolysis may have a significant impact on the reliability and quality of the results. Our experimental evidence suggests that the risk of false-negative results can be minimized by (i) the expansion of the antigen panel, with the inclusion of four additional targets; (ii) a more accurate selection of the gating area of the red blood cells; (iii) the choice of a better fluorochrome (alexa fluor 488) to be conjugated to the secondary antibody; and (iv) the implementation of different staining protocols depending on the nature of the double population to be detected (donor expressing vs. recipient non-expressing and vice versa). The combination of the above approaches allowed a significant reduction of false-negative results, assessed on samples simulating a homologous blood transfusion between two compatible subjects.
本文介绍了一项研究的结果，旨在为改进基于流式细胞荧光术的方法在兴奋剂检测中检测同源输血的实施提供新的建议和策略。该方法基于识别供体和受体所拥有的少数血型抗原之间的表型不匹配。为降低假阴性结果的风险，采取了两种策略：（i）监测更广泛的红细胞表面抗原；（ii）根据不同抗原以及必须检测的抗原不匹配类型（无论是供体还是受体表达或不表达待检测抗原）应用不同的红细胞表面染色方案。还特别关注了时间因素，以避免样本长时间储存，因为溶血可能对结果的可靠性和质量产生重大影响。我们的实验证据表明，通过以下方式可将假阴性结果的风险降至最低：（i）扩大抗原组，增加四个额外靶点；（ii）更准确地选择红细胞的设门区域；（iii）选择更好的荧光染料（alexa fluor 488）与二抗结合；（iv）根据待检测的双群体性质（供体表达与受体不表达以及反之亦然）实施不同的染色方案。上述方法的组合显著减少了假阴性结果，这是在模拟两个相容受试者之间同源输血的样本上评估得出的。
