Úlehlová Jana, Piskláková Barbora, Ivanovová Eliška, Procházková Jana, Bradáčová Pavla, Kvasnička Aleš, Friedecký David, Slavík Luděk
Department of Hematology-Oncology, Faculty of Medicine and Dentistry, Palacký University and University Hospital Olomouc, 779 00 Olomouc, Czech Republic.
Laboratory for Inherited Metabolic Disorders, Department of Clinical Biochemistry, Palacký University and University Hospital Olomouc, 779 00 Olomouc, Czech Republic.
Diagnostics (Basel). 2021 Nov 2;11(11):2027. doi: 10.3390/diagnostics11112027.
The effect of direct oral anticoagulants (DOAC) on laboratory tests dependent on the production of their targets, factor IIa and factor Xa, is a well-known problem and can cause both false positive and negative results. In particular, the situation in patients who develop lupus anticoagulant (LA) antibodies is highly complex. To evaluate the effectiveness of DOAC therapy in lupus-positive patients, 31 samples were enrolled in this retrospective study. All patient samples were spiked with three types of DOAC (dabigatran, DABI; rivaroxaban, RIVA; and apixaban, API) in a concentration that significantly influenced the screening test for LA and thus can mask the presence of LA. Subsequently, the DOAC was always unbound by the DOAC-Stop procedure. DOAC levels before and after binding were determined by functional assays, followed by liquid chromatography coupled with mass spectrometry (LC-MS) analysis.
The determination of DOAC levels was performed by direct thrombin assay and determination of anti-Xa activity with specific calibration as functional tests for DABI and xabans (API and RIVA). To determine concentration levels of API, DABI, and RIVA, our in-house LC-MS method was used.
The results of LA-positive samples show significant differences between functional tests and the LC-MS method both before and after DOAC binding.
The acute findings of the presence of LA-type antibodies fundamentally affects the determination of DOAC by functional tests, and in this case, it is necessary to use LC-MS analysis to determine the true value. If patients treated with DOAC develop LA of medium and higher titers, we do not recommend checking DOAC levels with functional tests.
直接口服抗凝剂（DOAC）对依赖其作用靶点（凝血因子IIa和Xa）生成的实验室检测的影响是一个众所周知的问题，可能导致假阳性和假阴性结果。特别是，出现狼疮抗凝物（LA）抗体的患者情况极为复杂。为评估DOAC疗法在狼疮阳性患者中的有效性，本回顾性研究纳入了31份样本。所有患者样本均加入三种类型的DOAC（达比加群，DABI；利伐沙班，RIVA；阿哌沙班，API），其浓度对LA筛查试验有显著影响，因此可能掩盖LA的存在。随后，通过DOAC-Stop程序使DOAC始终处于未结合状态。通过功能测定法测定结合前后的DOAC水平，随后进行液相色谱-质谱联用（LC-MS）分析。
通过直接凝血酶测定法和采用特定校准的抗Xa活性测定法来测定DOAC水平，作为DABI和沙班类药物（API和RIVA）的功能测试。为测定API、DABI和RIVA的浓度水平，使用了我们内部的LC-MS方法。
LA阳性样本的结果显示，在DOAC结合前后，功能测试和LC-MS方法之间均存在显著差异。
LA型抗体存在的急性发现从根本上影响了通过功能测试对DOAC的测定，在这种情况下，有必要使用LC-MS分析来确定真实值。如果接受DOAC治疗的患者出现中高滴度的LA，我们不建议使用功能测试来检查DOAC水平。
