Takizawa M, Maguchi S, Ryu T, Oridate N, Fukuda S, Inuyama Y, Honke K
Department of Otorhinolaryngology, Hokkaido University School of Medicine, Sapporo.
Nihon Jibiinkoka Gakkai Kaiho. 1994 Jan;97(1):20-7. doi: 10.3950/jibiinkoka.97.20.
Anti-neutrophil cytoplasmic antibodies (ANCA) have been reported to be a disease-specific marker for Wegener's granulomatosis (WG). In the present study we developed an enzyme-linked immunosorbent assay (ELISA) for detecting and quantifying ANCA. The cytoplasmic alpha-fraction of neutrophils obtained from healthy human donors was used as an antigen. The alpha-fraction was purified from supernatants of homogenized neutrophils by sucrose gradient centrifugation. Peroxidase conjugated rabbit anti-human IgG was used as a secondary antibody. Diluted sera from 13 patients with WG, 15 with Sjögren syndrome, 1 with polymorphic reticulosis, 3 with relapsing polychondritis, 4 with other collagen diseases, 4 with sinusitis and 18 healthy donors were examined. It was concluded that this ELISA was sufficiently specific and sensitive for WG, and the ELISA units correlated individually with the ANCA titers as determined by an indirect immunofluorescence technique. This ELISA provides precise ANCA quantitation and will be useful for the diagnosis of WG and for monitoring its activity.
抗中性粒细胞胞浆抗体（ANCA）据报道是韦格纳肉芽肿病（WG）的一种疾病特异性标志物。在本研究中，我们开发了一种用于检测和定量ANCA的酶联免疫吸附测定（ELISA）。从健康人类供体获得的中性粒细胞的胞浆α组分用作抗原。α组分通过蔗糖梯度离心从匀浆化中性粒细胞的上清液中纯化得到。过氧化物酶偶联的兔抗人IgG用作二抗。检测了13例WG患者、15例干燥综合征患者、1例多形性网状细胞增多症患者、3例复发性多软骨炎患者、4例其他胶原病患者、4例鼻窦炎患者以及18名健康供体的稀释血清。得出的结论是，该ELISA对WG具有足够的特异性和敏感性，且ELISA单位与通过间接免疫荧光技术测定的ANCA滴度呈个体相关。这种ELISA可提供精确的ANCA定量，将有助于WG的诊断及其活动监测。
