Roback John D, Barclay Sheilagh, Hillyer Christopher D
Transfusion Medicine Program, Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia, USA.
Transfusion. 2003 Jul;43(7):918-27. doi: 10.1046/j.1537-2995.2003.t01-1-00433.x.
Current immunohematology testing methods have limitations including cost, throughput, and adaptability to automation. Furthermore, current automated and semiautomated workstations cannot accommodate many other tests relevant to blood transfusion.
Authentic clinical samples from hospitalized patients were tested for ABO group, D type, and presence of RBC alloantibodies by column agglutination technology (CAT), standard tube methods, and a recently developed flow cytometry (FC) technique. Included were challenging samples with rouleaux, autoantibodies, mixed-field reactions, and weak antibodies. Antibody staining of RBCs for FC was initially performed in test tubes and subsequently in microtiter filter plates interfaced with a vacuum manifold.
When antibody staining was performed in tubes, FC testing determined the correct ABO group and D type for 99.1 percent of 222 clinical samples, as compared to accuracies of 91.9 percent for CAT and 95.0 percent for standard tube testing. FC testing also detected 99.5 percent of clinically relevant RBC alloantibodies in 239 patient samples, as compared to 98.9 percent for CAT and 94.7 percent for LISS-IAT. Using the FC filter plate technique, 104 of 109 samples (95.4%) were correctly typed for ABO and D (the remaining five samples were read as "no type determined" due to RBC and serum testing discrepancies), and RBC alloantibodies of the IgG and IgM classes were correctly identified in 98.3 percent of samples.
Optimized FC testing methods that are comparable in accuracy to standard CAT and tube methods are described. When used with filter plates, this methodology should allow rapid and cost-effective immunohematology testing of both patient and donor samples in an automated workstation format. The same workstation should support automation of other pretransfusion assays that can be analyzed by FC.
当前免疫血液学检测方法存在局限性，包括成本、通量以及对自动化的适应性。此外，当前的自动化和半自动化工作站无法进行许多与输血相关的其他检测。
采用柱凝集技术（CAT）、标准试管法以及最近开发的流式细胞术（FC）技术，对住院患者的真实临床样本进行ABO血型、D抗原类型及红细胞同种抗体检测。检测样本包括存在缗钱状形成、自身抗体、混合视野反应及弱抗体的挑战性样本。红细胞的抗体染色最初在试管中进行，随后在与真空歧管连接的微量滴定滤板中进行。
当在试管中进行抗体染色时，FC检测对222份临床样本中99.1%的样本确定了正确的ABO血型和D抗原类型，相比之下，CAT的准确率为9l.9%，标准试管检测的准确率为95.0%。FC检测还在239份患者样本中检测出99.5%的临床相关红细胞同种抗体，相比之下，CAT为98.9%，低离子强度盐水-抗球蛋白试验（LISS-IAT）为94.7%。采用FC滤板技术，109份样本中的104份（95.4%）ABO血型和D抗原类型鉴定正确（其余5份样本因红细胞和血清检测结果不一致而判读为“未确定血型”），98.3%的样本中正确鉴定出IgG和IgM类红细胞同种抗体。
描述了优化后的FC检测方法，其准确性与标准CAT和试管法相当。当与滤板一起使用时，该方法应能在自动化工作站形式下对患者和供体样本进行快速且经济高效的免疫血液学检测。同一工作站应支持可通过FC分析的其他输血前检测的自动化。
