Related Experiment Video
Updated: May 12, 2026
13:01
Anti-Nuclear Antibody Screening Using HEp-2 Cells
Published on: June 23, 2014
Automated indirect immunofluorescence antinuclear antibody analysis is a standardized alternative for visual
Clinical Chemistry and Laboratory Medicine
|April 25, 2013
Summary
Automated antinuclear antibody (ANA) indirect immunofluorescence (IIF) analysis shows high agreement with manual methods for detection and intensity but limited pattern recognition. Expert interpretation remains crucial for accurate ANA diagnostics.
Area of Science:
- Immunology
- Rheumatology
- Clinical Diagnostics
Background:
- Antinuclear antibody (ANA) screening is essential for diagnosing systemic rheumatic disorders.
- Indirect immunofluorescence (IIF) is the gold standard for ANA detection, despite standardization challenges.
- Automated IIF offers a potential standardized alternative to manual ANA analysis.
Purpose of the Study:
- To evaluate the reliability of automated ANA IIF analysis (Zenit G-sight) compared to conventional methods.
- To assess the performance of automated ANA IIF in detecting ANA and identifying disease-specific patterns.
- To determine the potential of automated systems to improve standardization in ANA testing.
Main Methods:
- 304 routine sera, 28 rare pattern samples, 219 disease cohort samples, and 100 healthy donor samples were analyzed.
- Samples were tested using automated IIF (Zenit G-sight), conventional visual IIF microscopy, and two ANA enzyme immunoassays (EIA).
- Comparison focused on negative/positive interpretation, intensity assessment, and pattern recognition accuracy.
Main Results:
- Automated and conventional ANA IIF demonstrated >90% agreement for negative/positive interpretation and intensity.
- Pattern recognition accuracy for automated IIF was limited at 26%.
- Zenit G-sight showed higher sensitivity for systemic sclerosis (SSc)-associated antibody subsets than EIA, with a significant correlation to end-point titers.
Conclusions:
- Automated ANA IIF analysis using Zenit G-sight can improve standardization in ANA testing.
- Expert technicians are still required for accurate pattern recognition and classification of ambiguous samples.
- Automated systems show promise but require complementary human expertise for comprehensive ANA diagnostics.
More Related Videos
Related Concept Videos
01:12Immunofluorescence Microscopy
A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
The...
The...
01:20Immunogold Electron Microscopy
Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
01:22Immunocytochemistry and Immunohistochemistry
Immunocytochemistry (ICC) and immunohistochemistry (IHC) are techniques that use antibodies to check for specific proteins or antigens in a sample. The technique was first published by Albert Coons in 1941 to detect the presence of pneumococcal antigen in tissue sections from mice infected with Pneumococcus. Immunocytochemistry helps localization of proteins or antigens in individual cells like blood cells, stem cells, etc., while immunohistochemistry does the same for tissue samples.
These...
These...