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Diagnostic value of Immunoblotting assay for determination of anti nuclear antibody (ANA) concentration in rheumatologic diseases

Mohammad Naghavi-Behzad1*
  • 1 Tabriz University of Medical Sciences, Students' Research Committee, Iran

Abstract
Background: Antinuclear antibodies (ANAs) are common features of autoimmune connective tissue diseases. Various detection methods are used and there are newer techniques that are continuously put forward to facilitate diagnosis and therapeutic monitoring in connective tissue disease (CTD) patients. Immunofluorescence (IF) or Fluorescent Antibody Technique the most used and “gold standard” test for diagnosis. Enzyme-Linked Immunosorbent Assay (ELISA) is another routine test. For Immunoblotting (IB) assay, autoimmune ANA profiles are used which provide a qualitative in vitro assay for human autoantibodies to 15 different antigens. This study was conducted to compare three techniques (IF, IB, ELISA) in detection of ANA. If the sensitivity and specificity of IB superior to other methods, we can replace IF and ELISA with IB.
Materials and Methods: An analytical cross-sectional study of 85 sera from patients with Systemic lupus erythematous (SLE), Systemic sclerosis (SSc) and Dermatomyosities(DM) was undertaken at rheumatology and nephrology department and clinic of Emam Reza hospital from 89/11/1 to 90/10/30. Sera collected and stored at -80˚c. Then they were used to detection of ANA with three techniques.
Results: Of all sera 63 (74.1%) were ELISA positive and 22 (25.9%) had negative ELISA.74 (87.1 %) IF and IB positive and 11 (12.9%) IF and IB negative were observed. The sensitivity and specificity of IB in comparison with IF was 98.65% and 90.91% respectively. In comparison with ELISA we found 93.65% and 31.82% of sensitivity and specificity.
Conclusion: Immunoblotting, has high sensitivity and specificity, can be used in screening of ANA.

References

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3. Aksu G, Gulez N, Azarsiz E, Karaka N, Kutukculer N. Determination of cut-off titers and agreement between immunofluorescence and immunoblotting methods for detecting antinuclear antibodies in children.Journal of Clinical Laboratory Analysis 2010; 24:230-236.

4. S.Copple S, D.Sawitzke A, M.Wilson A, E.Tebo A, R.Hill H. Enzyme-Linked Immunosorbent Assay screening then Indirect Immunofluorescence confirmation of Antinuclear Antibodies.Am J ClinPathol 2011;135(4):678-684.

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Keywords: Enzyme-Linked Immunosorbent Assay, Immunoblotting, Fluorescent Antibody Technique, Connective Tissue Diseases, rheumatologic diseases

Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.

Presentation Type: Abstract

Topic: Immune-mediated disease pathogenesis

Citation: Naghavi-Behzad M (2013). Diagnostic value of Immunoblotting assay for determination of anti nuclear antibody (ANA) concentration in rheumatologic diseases. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00757

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Received: 16 Jun 2013; Published Online: 22 Aug 2013.

* Correspondence: Dr. Mohammad Naghavi-Behzad, Tabriz University of Medical Sciences, Students' Research Committee, Tabriz, Iran, mohammad_nb@yahoo.com