Efficient Detection of Anti-Nuclear Antibodies
University of Wales Institute Cardiff
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A progressively wide range of studies exist into the association of Anti-Nuclear Antibodies (ANAs) with Autoimmune Rheumatic Diseases (ARDS), such as Systemic Lupus Erythematosus (SLE), Sjogrens Syndrome (SjS), Systemic Scleroderma (SS), Rheumatoid Arthritis (RA), Reynaud’s phenomena (RP) to name but a few of the more serious and debilitating ARDs1-4. It is thought that a single base mutation in the gene responsible for anti-Phosphotidylcholine (aPTC), (an IgM Natural Autoantibody (NAb) responsible for clearance of cellular debris) is a likely mechanism by which ANA pathogenesis may originate5. Visualization of these antibodies through Indirect Immuno-Fluorescence (IIF) has long remained the principle detection path, the “Gold Standard”. The objective of this study is to investigate the replacement of IIF with ELISA as a screening/diagnostic method in the ongoing effort to eliminate the time-consuming and subjective nature of IIF; ELISA being easily adapted to automation. 350 routine anonymised patient samples were prepared and tested in the Immunology laboratory, Singleton Hospital, by IIF and ELISA. IIF samples were prepared on the Beeline 220s upon Hep-2 laryngeal carcinoma cell slides, with further analysis being carried out through fluorescence microscopy. Results gathered from initial IIF analysis were checked for accuracy by two, more experienced biomedical scientists. The “Best 2000” Biokit ELISA system prepared, tested and analysed ELISA patient samples. Investigation of results was carried out through chi-squared analyses of result association between the tests. Findings revealed significant association (a p value of <0.0005) between the presence of a positive result and the test used. Investigation of patterns missed by ELISA test was also investigated, showing 50 – 76% of clinically significant patterns were missed. Epitope integrity and occlusion are thought to results in the lower sensitivity and specificity of ELISA testing. Therefore illustrating conclusions that the role of ELISA in ANA detection be limited to disease management and response to treatment.
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