Jacobs Jeremy W, Stuart Melissa S, Tange Julie I, Leger Rachel R, Ashrani Aneel A, Chen Dong, Pruthi Rajiv K, Sridharan Meera, Seheult Jansen N
Department of Pathology, Microbiology & Immunology, Vanderbilt University Medical Center, Nashville, TN, US.
Special Coagulation Laboratory, Division of Hematopathology, Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, US.
Am J Clin Pathol. 2025 Apr 19;163(4):610-617. doi: 10.1093/ajcp/aqae160.
Fluorescence resonance energy transfer (FRET)-based ADAMTS13 activity assays are critical for the diagnosis of thrombotic thrombocytopenic purpura. However, these assays are susceptible to interference. As iodide has been suggested to interfere in laboratory testing via fluorophore quenching or promotion, we aimed to determine whether iodinated contrast (Omnipaque) interferes with the ATS-13 ADAMTS13 Activity Assay 2.0.
We evaluated the excitation, emission, and absorbance spectrum of Omnipaque alone and spiked in patient plasma with known ADAMTS13 activity and ADAMTS13 activity on Omnipaque alone, an abnormal control of patient plasma previously observed to display elevated baseline relative fluorescent units, and variable concentrations of patient plasma with known ADAMTS13 activity spiked with Omnipaque.
No atypical fluorescent peaks were observed on any sample (Omnipaque alone or spiked in plasma) between 250 and 700 nm. There was no difference in the mean ADAMTS13 activity among the various concentrations of plasma spiked with Omnipaque or plasma spiked with saline.
Iodinated contrast does not appear to interfere-either via spectral overlap of the fluorophore or through physiologic inhibition of the ADAMTS13 enzyme-with ADAMTS13 activity FRET-based assays based on the findings from this in vitro analysis. Delaying sample collection for ADAMTS13 activity testing from suspected patients with thrombotic thrombocytopenic purpura following administration of iodinated radiocontrast agents is not necessary, and recent contrast administration should not yield erroneous ADAMTS13 activity results.
基于荧光共振能量转移(FRET)的ADAMTS13活性检测对于血栓性血小板减少性紫癜的诊断至关重要。然而,这些检测易受干扰。由于有人提出碘化物可通过荧光团猝灭或促进作用干扰实验室检测,我们旨在确定碘化造影剂(欧乃派克)是否会干扰ATS - 13 ADAMTS13活性检测2.0。
我们评估了单独的欧乃派克以及加入已知ADAMTS13活性的患者血浆中的欧乃派克的激发光谱、发射光谱和吸收光谱,还评估了单独的欧乃派克的ADAMTS13活性、先前观察到基线相对荧光单位升高的患者血浆异常对照以及加入欧乃派克的不同浓度已知ADAMTS13活性的患者血浆的ADAMTS13活性。
在250至700纳米之间,任何样品(单独的欧乃派克或加入血浆中的)均未观察到非典型荧光峰。加入欧乃派克的不同浓度血浆或加入生理盐水的血浆之间,平均ADAMTS13活性没有差异。
基于这项体外分析的结果,碘化造影剂似乎不会通过荧光团的光谱重叠或ADAMTS13酶的生理抑制作用干扰基于FRET的ADAMTS13活性检测。对于疑似血栓性血小板减少性紫癜患者,在给予碘化放射性造影剂后,无需延迟采集用于ADAMTS13活性检测的样本,近期使用造影剂也不应产生错误的ADAMTS13活性结果。