Paspaliaris B, Pamio M, Savige J
Department of Biochemistry, St Vincent's Hospital, Fitzroy, VIC, Australia.
J Clin Pathol. 2000 Oct;53(10):774-7. doi: 10.1136/jcp.53.10.774.
The "International consensus document on testing and reporting of antineutrophil cytoplasmic antibodies (ANCA)" requires all sera to be examined by indirect immunofluorescence (IIF). However, commercial neutrophil slides are expensive, fluorescence patterns can be difficult to interpret, and coincidental antinuclear antibodies (ANA) cannot be demonstrated; in addition, in house cytospin neutrophil preparations are time consuming to prepare and deteriorate with time.
To compare the IIF demonstration of ANCA, using washed peripheral blood cell smears, with commercial neutrophil preparations and with ANCA positivity as demonstrated by enzyme linked immunosorbent assay (ELISA).
Serum fluorescence positivity, pattern, and intensity using washed peripheral blood cell smears were compared with the results obtained using commercial neutrophil slides (INOVA). Fluorescence positivity, pattern, and intensity of 500 sera from consecutive patients with suspected vasculitis tested with washed peripheral blood cells were compared with binding in ELISAs for proteinase 3 (PR3) and myeloperoxidase (MPO).
IIF of washed peripheral blood cell smears detected seven of eight sera with cytoplasmic fluorescence (C-ANCA), and 11 of 12 sera with perinuclear fluorescence (P-ANCA) demonstrated using commercial slides. The two sera that were negative by IIF were also negative in the ELISAs for both PR3-ANCA and MPO-ANCA. Of the 500 sera examined, there were 35 (7%) with C-ANCA, 65 (13%) with P-ANCA, and eight (2%) IIF negative sera that were positive by either ELISA. There was a strong correlation between C-ANCA fluorescence and PR3-ANCA values (p < 0.0001), and a moderate to strong correlation between P-ANCA fluorescence and MPO-ANCA values (p < 0.001) when ANCA fluorescence was demonstrated with washed peripheral blood cell smears.
Washed peripheral blood cells are a convenient and useful low cost alternative to commercial or cytospin neutrophil preparations for the IIF demonstration of ANCA.
“抗中性粒细胞胞浆抗体（ANCA）检测与报告国际共识文件”要求所有血清均采用间接免疫荧光法（IIF）检测。然而，市售中性粒细胞玻片价格昂贵，荧光模式可能难以解读，且无法显示同时存在的抗核抗体（ANA）；此外，自制的细胞离心涂片中性粒细胞制剂制备耗时且会随时间变质。
比较使用洗涤后的外周血细胞涂片进行ANCA的IIF检测结果与使用市售中性粒细胞制剂的检测结果，以及与酶联免疫吸附测定（ELISA）所显示的ANCA阳性结果。
将使用洗涤后的外周血细胞涂片检测的血清荧光阳性率、模式及强度与使用市售中性粒细胞玻片（INOVA）获得的结果进行比较。将连续的500例疑似血管炎患者的血清用洗涤后的外周血细胞检测的荧光阳性率、模式及强度与蛋白酶3（PR3）和髓过氧化物酶（MPO）的ELISA结合情况进行比较。
使用洗涤后的外周血细胞涂片进行IIF检测，检测出了使用市售玻片显示为胞浆荧光（C-ANCA）的8份血清中的7份，以及核周荧光（P-ANCA）显示为阳性的12份血清中的11份。IIF检测为阴性的2份血清在PR3-ANCA和MPO-ANCA的ELISA检测中也为阴性。在检测的500份血清中，有35份（7%）为C-ANCA阳性，65份（13%）为P-ANCA阳性，8份（2%）IIF检测为阴性的血清在ELISA检测中呈阳性。当使用洗涤后的外周血细胞涂片显示ANCA荧光时，C-ANCA荧光与PR3-ANCA值之间存在强相关性（p<0.0001），P-ANCA荧光与MPO-ANCA值之间存在中度至强相关性（p<0.001）。
对于ANCA的IIF检测，洗涤后的外周血细胞是一种方便、有用且低成本的替代市售或细胞离心涂片中性粒细胞制剂的方法。
