Katsumata Yasuhiro, Sada Ken-Ei, Kameda Tomohiro, Dobashi Hiroaki, Kaname Shinya, Tsuboi Naotake, Matsumoto Yoshinori, Amano Koichi, Tamura Naoto, Harigai Masayoshi
Division of Rheumatology, Department of Internal Medicine, Tokyo Women's Medical University School of Medicine, Tokyo, Japan.
Department of Clinical Epidemiology, Kochi Medical School, Nankoku, Japan.
Immunol Med. 2025 Mar;48(1):47-57. doi: 10.1080/25785826.2024.2408054. Epub 2024 Oct 11.
We compared different antineutrophil cytoplasmic antibody (ANCA) detection methods using a predominantly myeloperoxidase (MPO)-ANCA-associated vasculitis cohort. Stored sera from 147 patients with untreated ANCA-associated vasculitis (AAV), including microscopic polyangiitis and granulomatosis with polyangiitis ( = 115 and 32, respectively), and 124 disease controls were tested for P-ANCA and C-ANCA with immunofluorescence (IIF), and for MPO-ANCA and proteinase 3 (PR3)-ANCA with different antigen-specific immunoassays: direct enzyme-linked immunosorbent assay (ELISA), chemiluminescent enzyme immunoassay (CLEIA), third-generation fluorescent enzyme immunoassay (FEIA), and latex turbidimetrical immunoassay (LTIA). In addition, MPO-ANCA and PR3-ANCA titers were calibrated using certified reference materials (CRMs). The sensitivities and specificities for AAV diagnoses were 95% and 94% (IIF), 86% and 98% (ELISA), 93% and 94% (CLEIA), 92% and 96% (FEIA), and 68% and 88% (LTIA). Dual IIF/antigen-specific immunoassay testing reduced diagnostic accuracies from 94% to 93%. The quantitative agreement between ANCA levels measured using CLEIA and FEIA and calibrated using CRMs was not good. In conclusion, this study demonstrated the high performance of antigen-specific immunoassays for AAV diagnosis in a predominantly MPO-ANCA-associated vasculitis cohort and suggested that the benefit of dual IIF/antigen-specific immunoassay testing is limited. Standardizing ANCA measurements using different immunoassays was difficult, even when using CRMs.
我们使用一个主要为髓过氧化物酶（MPO）-抗中性粒细胞胞浆抗体（ANCA）相关血管炎的队列，比较了不同的ANCA检测方法。对147例未经治疗的ANCA相关血管炎（AAV）患者（包括显微镜下多血管炎和肉芽肿性多血管炎患者，分别为115例和32例）以及124例疾病对照的储存血清，采用免疫荧光法（IIF）检测P-ANCA和C-ANCA，并采用不同的抗原特异性免疫测定法检测MPO-ANCA和蛋白酶3（PR3）-ANCA：直接酶联免疫吸附测定（ELISA）、化学发光酶免疫测定（CLEIA）、第三代荧光酶免疫测定（FEIA）和乳胶比浊免疫测定（LTIA）。此外，使用认证参考物质（CRM）对MPO-ANCA和PR3-ANCA滴度进行校准。AAV诊断的敏感性和特异性分别为95%和94%（IIF）、86%和98%（ELISA）、93%和94%（CLEIA）、92%和96%（FEIA）以及68%和88%（LTIA）。IIF/抗原特异性免疫测定双重检测使诊断准确性从94%降至93%。使用CLEIA和FEIA测量并经CRM校准的ANCA水平之间的定量一致性不佳。总之，本研究证明了抗原特异性免疫测定在一个主要为MPO-ANCA相关血管炎队列中对AAV诊断的高性能，并表明IIF/抗原特异性免疫测定双重检测的益处有限。即使使用CRM，使用不同免疫测定法标准化ANCA测量也很困难。
