Rigon Amelia, Soda Paolo, Zennaro Danila, Iannello Giulio, Afeltra Antonella
Department of Clinical Medicine, Immunology, and Rheumatology, University Campus Bio-Medico, Rome, Italy.
Cytometry B Clin Cytom. 2007 Nov;72(6):472-7. doi: 10.1002/cyto.b.20356.
The recommended method for antinuclear antibodies (ANA) detection is indirect immunofluorescence (IIF). To pursue a high image quality without artefacts and reduce interobserver variability, this study aims at evaluating the reliability of automatically acquired digital images of IIF slides for diagnostic purposes.
Ninety-six sera were screened for ANA by IIF on HEp-2 cells. Two expert physicians looking at both the fluorescence microscope and the digital images on computer monitor performed a blind study to evaluate fluorescence intensity and staining pattern. Cohen's kappa was used as an agreement evaluator between methods and experts.
Considering fluorescence intensity, there is a substantial agreement between microscope and monitor analysis in both physicians. Agreement between physicians was substantial at the microscope and perfect at the monitor. Considering IIF pattern, there was a substantial and moderate agreement between microscope and monitor analysis in both physicians. Kappa between physicians was substantial both at the microscope and at the monitor.
These preliminary results suggest that digital media is a reliable tool to help physicians in detecting autoantibodies in IIF. Our data represent a first step to validate the use of digital images, thus offering an opportunity for standardizing and automatizing the detection of ANA by IIF.
抗核抗体（ANA）检测的推荐方法是间接免疫荧光法（IIF）。为了获得高质量且无伪影的图像并减少观察者间的差异，本研究旨在评估用于诊断目的的IIF载玻片自动采集数字图像的可靠性。
采用IIF法在人喉表皮样癌细胞（HEp-2细胞）上对96份血清进行ANA筛查。两名专家医生在荧光显微镜和计算机显示器上查看数字图像，进行盲法研究以评估荧光强度和染色模式。Cohen's kappa系数用作方法与专家之间的一致性评估指标。
在荧光强度方面，两位医生在显微镜分析和显示器分析之间均有高度一致性。两位医生在显微镜下的一致性高度显著，在显示器上的一致性完美。在IIF模式方面，两位医生在显微镜分析和显示器分析之间均有高度和中度一致性。两位医生在显微镜和显示器上的kappa系数均高度显著。
这些初步结果表明，数字媒体是帮助医生在IIF中检测自身抗体的可靠工具。我们的数据是验证数字图像使用的第一步，从而为通过IIF标准化和自动化ANA检测提供了机会。
