流式细胞技术检测人异体血液回输方法研究  

Detection of Human Homologous Blood Transfusion by Flow Cytometer

作　　者：河春姬[1] 杨声[1] 董颖[1] 张力思[1] 景晶[1] 徐友宣[1] 吴侔天[1] 

机构地区：[1]国家体育总局反兴奋剂中心,北京100029

出　　处：《体育科学》2015年第1期93-96,F0003,共5页China Sport Science

摘　　要：目的：血液回输能够迅速增加机体红细胞数，增强有氧运动能力，提高运动成绩。采用流式细胞技术对异体血液回输的检测方法进行适用性验证研究，以证明该技术检测异体血液回输的可靠性和可操作性。研究方法：应用流式细胞技术，通过检测血液中荧光标记的红细胞血型抗原表达形式的方法，追踪是否存在微量的异体红细胞群，并判断是否接受异体血液回输。3名检测员使用流式细胞仪对46例不同比例（0～5％）体外混合的全血进行8种红细胞血型抗原的检测。结果：本研究对该方法的特异性、灵敏度、精密度、稳定性等进行评估。此方法检测无假阳性结果；可准确检测不低于3％的异体输血量，灵敏度较高；经细胞稳定液处理4～8℃储存的样品在4周内重复检测结果一致，3名检测员检测的结果一致。结论：流式细胞技术检测异体血液回输的方法准确可靠，符合实验室国际标准（ISO-17025）认证和验证要求，已通过中国合格评定认可委员会认可，并应用于常规和奥运会等大型赛事的异体血液回输兴奋剂检测。Objective ： Blood transfusion could increase red blood cells （RBCs） rapidly and may enhance endurance performance significantly. A validation was carried out to prove the reliability and operability of the detection of homologous blood transfusion with flow cytometry technology. Method.. Homologous blood transfusion was detected using flow cytometry technique to observe the presence of different RBCs phenotypes of eight different antigens in a blood sample. The analysis of 46 blood samples containing different percentages （0 - 5 %） of homologous RBCs was carried out by three independent analysts. Results：The method afforded satisfactory results in terms of robustness, sensitivity, specificity, precision and stability. No false positive results were observed. All samples contained 3 % homologous RBCs were unambiguously detected. These samples were stable over 4 weeks after using cell stable buffer at 4 8℃. Conclusion： This method fulfils the ISO-17025 accreditation and has been accredited by China National Accreditation Service for Conformity Assessment （CNAS）. The method has been successfully utilized to the detection of homologous blood transfusion in major events and routine doping-control samples.

关 键 词：异体血液回输 血型抗原 流式细胞仪 方法验证 

分 类 号：G804.7[文化科学—运动人体科学]