Subscribe to RSS
Download PDF
DOI: 10.1055/s-0041-1731945
A Comparative Evaluation of Antigen-Specific Sandwich Immunoassay and Indirect Immunofluorescence Assay (IIF) in Detecting Antineutrophil Cytoplasmic Antibodies: Are We Ready to Replace IIF with ELISA as the Primary Screening Method?
Abstract
Background Antineutrophil cytoplasmic antibodies (ANCA) are important biomarkers in the diagnosis of ANCA-associated vasculitis, and indirect immunofluorescence (IIF) had been the method of choice for its detection from the very beginning. However, international consensus on ANCA testing (2017) advocates the use of high-quality immunoassays as the primary screening method. The purpose of this study was to evaluate the diagnostic performance of enzyme-linked immunosorbent assay (ELISA) compared to IIF in detecting ANCA.
Methods One-hundred eighty-nine serum samples of suspected or known cases of systemic vasculitis were screened for ANCA by IIF and proteinase-3- and myeloperoxidase-ELISA. In IIF, positive results were further divided into cytoplasmic pattern of ANCA and perinuclear pattern of ANCA, depending upon the pattern of fluorescence. McNemar’s chi-squared test was applied to check the equality of proportions of positive results, and Kappa statistics was used to measure the agreement between the two methods. Diagnostic performance of ELISA was evaluated taking IIF as reference.
Results IIF detected ANCA in 17.5% cases and ELISA detected it in 11.6% cases. A good agreement between the overall performance of ELISA and IIF was observed (K-value: 6.8, p-value: < 001). However, a significant difference in the proportion of positive results by the two methods was observed in McNemar’s test (two-sided p-value: 0.007). Taking IIF as standard, ELISA showed 60.6% sensitivity, 98.7% specificity, and predictive value of positive and negative results of 90.9 and 92.2%, respectively.
Conclusion The new generation antigen-specific ELISAs had high specificity but the chances of missing cases in primary screening due to the low sensitivity and high false negativity (39.4%) need to be dealt with.
Publication History
Article published online:
09 July 2021
© 2021. The Indian Association of Laboratory Physicians. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial-License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/).
Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India
-
References
- 1 Csernok E, Holle JU. Twenty-eight years with antineutrophil cytoplasmic antibodies (ANCA): how to test for ANCA - evidence-based immunology?. Auto Immun Highlights 2010; 1 (01) 39-43
- 2 Csernok E, Lamprecht P, Gross WL. Diagnostic significance of ANCA in vasculitis. Nat Clin Pract Rheumatol 2006; 2 (04) 174-175
- 3 Savige JA, Paspaliaris B, Silvestrini R. et al A review of immunofluorescent patterns associated with antineutrophil cytoplasmic antibodies (ANCA) and their differentiation from other antibodies. J Clin Pathol 1998; 51 (08) 568-575
- 4 Davies DJ, Moran JE, Niall JF, Ryan GB. Segmental necrotising glomerulonephritis with antineutrophil antibody: possible arbovirus aetiology?. Br Med J (Clin Res Ed) 1982; 285 (6342) 606
- 5 Bossuyt X, Cohen Tervaert JW, Arimura Y. et al Position paper: Revised 2017 international consensus on testing of ANCAs in granulomatosis with polyangiitis and microscopic polyangiitis. Nat Rev Rheumatol 2017; 13 (11) 683-692
- 6 Csernok E, Moosig F. Current and emerging techniques for ANCA detection in vasculitis. Nat Rev Rheumatol 2014; 10 (08) 494-501
- 7 Sowa M, Grossmann K, Knütter I. et al PLOS ONE Staff. Correction: simultaneous automated screening and confirmatory testing for vasculitis-specific ANCA. PLoS One 2015; 10 (03) e0120626
- 8 Allard-Chamard H, Liang P. Antineutrophil cytoplasmic antibodies testing and interpretation. Clin Lab Med 2019; 39 (04) 539-552
- 9 Damoiseaux J, Csernok E, Rasmussen N. et al Detection of antineutrophil cytoplasmic antibodies (ANCAs): a multicentre European Vasculitis Study Group (EUVAS) evaluation of the value of indirect immunofluorescence (IIF) versus antigen-specific immunoassays. Ann Rheum Dis 2017; 76 (04) 647-653
- 10 Kirkwood BR, Sterne AC. Analysis of binary outcomes. In: Medical Statistics. 2nd edition. Malden, MA: Wiley Blackwell 2009
- 11 Savige J, Gillis D, Benson E. et al International consensus statement on testing and reporting of antineutrophil cytoplasmic antibodies (ANCA). Am J Clin Pathol 1999; 111 (04) 507-513
- 12 Menezes JM, Rossener R, Silva APMD, Rodrigues SS, Mangueira CLP. Comparison between enzyme-linked immunosorbent assay and indirect immunofluorescence for detection of antineutrophil cytoplasmic antibodies. Einstein (Sao Paulo) 2020; 18: eAO5132 (São Paulo)
- 13 Schulte-Pelkum J, Radice A, Norman GL. et al Novel clinical and diagnostic aspects of antineutrophil cytoplasmic antibodies. J Immunol Res 2014; 2014: 185416
- 14 Rao DA, Wei K, Merola JF. et al Myeloperoxidase-antineutrophil cytoplasmic antibodies (MPO-ANCA) and proteinase 3-ANCA without immunofluorescent ANCA found by routine clinical testing. J Rheumatol 2015; 42 (05) 847-852
- 15 Harris A, Chang G, Vadas M, Gillis D. ELISA is the superior method for detecting antineutrophil cytoplasmic antibodies in the diagnosis of systemic necrotising vasculitis. J Clin Pathol 1999; 52 (09) 670-676
- 16 Stone JH, Talor M, Stebbing J. et al Test characteristics of immunofluorescence and ELISA tests in 856 consecutive patients with possible ANCA-associated conditions. Arthritis Care Res 2000; 13 (06) 424-434