Indirect ImmunoFluorescence (IIF) assays are recommended as the gold standard method for detection of antinuclear antibodies (ANAs), which are of considerable importance in the diagnosis of autoimmune diseases. Fluorescence intensity analysis is very often complex, and depending on the capabilities of the operator, the association with incorrect classes is statistically easy. In this paper, we present a Convolutional Neural Network (CNN) system to classify positive/negative fluorescence intensity of HEp-2 IIF images, which is important for autoimmune diseases diagnosis. The method uses the best known pre-trained CNNs to extract features and a support vector machine (SVM) classifier for the final association to the positive or negative classes. This system has been developed and the classifier was trained on a database implemented by the AIDA (AutoImmunité, Diagnostic Assisté par ordinateur) project. The method proposed here has been tested on a public part of the same database, consisting of 2080 IIF images. The performance analysis showed an accuracy of fluorescent intensity around 93%. The results have been evaluated by comparing them with some of the most representative state-of-the-art works, demonstrating the quality of the system in the intensity classification of HEp-2 images.

Antinuclear antibodies (ANAs) are significant biomarkers in the diagnosis of autoimmune diseases in humans, done by mean of Indirect ImmunoFluorescence (IIF) method, and performed by analyzing patterns and fluorescence intensity. This paper introduces the AIDA Project (autoimmunity: diagnosis assisted by computer) developed in the framework of an Italy-Tunisia cross-border cooperation and its preliminary results. A database of interpreted IIF images is being collected through the exchange of images and double reporting and a Gold Standard database, containing around 1000 double reported images, has been settled. The Gold Standard database is used for optimization of a CAD (Computer Aided Detection) solution and for the assessment of its added value, in order to be applied along with an Immunologist as a second Reader in detection of autoantibodies. This CAD system is able to identify on IIF images the fluorescence intensity and the fluorescence pattern. Preliminary results show that CAD, used as second Reader, appeared to perform better than Junior Immunologists and hence may significantly improve their efficacy; compared with two Junior Immunologists, the CAD system showed higher Intensity Accuracy (85,5% versus 66,0% and 66,0%), higher Patterns Accuracy (79,3% versus 48,0% and 66,2%), and higher Mean Class Accuracy (79,4% versus 56,7% and 64.2%).

The search for anti-nucleus antibodies (ANA) represents a fundamental step in the diagnosis of autoimmune diseases. The test considered the gold standard for ANA research is indirect immunofluorescence (IIF). The best substrate for ANA detection is provided by Human Epithelial type 2 (HEp-2) cells. The first phase of HEp-2 type image analysis involves the classification of fluorescence intensity in the positive/negative classes. However, the analysis of IIF images is difficult to perform and particularly dependent on the experience of the immunologist. For this reason, the interest of the scientific community in finding relevant technological solutions to the problem has been high. Deep learning, and in particular the Convolutional Neural Networks (CNNs), have demonstrated their effectiveness in the classification of biomedical images. In this work the efficacy of the CNN fine-tuning method applied to the problem of classification of fluorescence intensity in HEp-2 images was investigated. For this purpose, four of the best known pre-trained networks were analyzed (AlexNet, SqueezeNet, ResNet18, GoogLeNet). The classifying power of CNN was investigated with different training modalities; three levels of freezing weights and scratch. Performance analysis was conducted, in terms of area under the ROC (Receiver Operating Characteristic) curve (AUC) and accuracy, using a public database. The best result achieved an AUC equal to 98.6% and an accuracy of 93.9%, demonstrating an excellent ability to discriminate between the positive/negative fluorescence classes. For an effective performance comparison, the fine-tuning mode was compared to those in which CNNs are used as feature extractors, and the best configuration found was compared with other state-of-the-art works.

The diagnosis and monitoring of autoimmune diseases are very important problem in medicine. The most used test for this purpose is the antinuclear antibody (ANA) test. An indirect immunofluorescence (IIF) test performed by Human Epithelial type 2 (HEp-2) cells as substrate antigen is the most common methods to determine ANA. In this paper we present an automatic HEp-2 specimen system based on a convolutional neural network method able to classify IIF images. The system consists of a module for features extraction based on a pre-trained AlexNet network and a classification phase for the cell-pattern association using six support vector machines and a k-nearest neighbors classifier. The classification at the image-level was obtained by analyzing the pattern prevalence at cell-level. The layers of the pre-trained network and various system parameters were evaluated in order to optimize the process. This system has been developed and tested on the HEp-2 images indirect immunofluorescence images analysis (I3A) public database. To test the generalisation performance of the method, the leave-one-specimen-out procedure was used in this work. The performance analysis showed an accuracy of 96.4% and a mean class accuracy equal to 93.8%. The results have been evaluated comparing them with some of the most representative works using the same database.