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Indirect immunofluorescence in autoimmune diseases: assessment of digital images for diagnostic purpose
Amelia Rigon1, Paolo Soda, Danila Zennaro
1Department of Clinical Medicine, Immunology, and Rheumatology, University Campus Bio-Medico, Rome, Italy. a.rigon@unicampus.it
Insights
Digital images of indirect immunofluorescence (IIF) slides reliably aid in detecting antinuclear antibodies (ANA). This study shows digital analysis offers substantial agreement, improving diagnostic consistency for autoimmune diseases.
Area of Science:
- Immunology
- Medical Diagnostics
- Digital Pathology
Background:
- Indirect immunofluorescence (IIF) is the standard for antinuclear antibodies (ANA) detection.
- High image quality and reduced interobserver variability are crucial for accurate ANA diagnostics.
- Evaluating automatically acquired digital IIF images is essential for diagnostic reliability.
Purpose of the Study:
- To assess the reliability of digital IIF images for ANA detection.
- To compare diagnostic performance using microscope versus digital image analysis.
- To explore standardization and automation in ANA detection.
Main Methods:
- Ninety-six sera were screened for ANA using IIF on HEp-2 cells.
- Two expert physicians conducted a blind study comparing microscope and digital image analysis.
- Cohen's kappa statistic evaluated agreement between methods and observers.
Main Results:
- Substantial agreement was observed between microscope and monitor analysis for fluorescence intensity.
- Physician agreement was substantial at the microscope and perfect at the monitor.
- Substantial and moderate agreement was found for IIF patterns between microscope and monitor analysis.
Conclusions:
- Digital images are a reliable tool for assisting physicians in detecting autoantibodies via IIF.
- This study is a foundational step towards validating digital image use in ANA diagnostics.
- Digital IIF image analysis presents opportunities for standardizing and automating ANA detection.
Background:
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.
Methods:
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.
Results:
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.
Conclusions:
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.
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