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Comparison of PR3-ANCA specific assay performance for the diagnosis of granulomatosis with polyangiitis (Wegener’s)

  • Antonella Radice EMAIL logo , Laura Bianchi , Umberto Maggiore , Augusto Vaglio and Renato A. Sinico
Published/Copyright: June 25, 2013
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 51 Issue 11

Abstract

Background: PR3-ANCA, the serological marker of granulomatosis with polyangiitis (GPA), is usually detected by immunometric assays, with purified PR3 directly coated onto the solid-phase. Novel methods for PR3-ANCA detection have been developed to improve the performance of traditional PR3-ANCA specific assays, but little is known about their diagnostic performance in real-life clinical settings. This study aimed to compare the performance of nine different commercial PR3-ANCA specific assays, including traditional and newer ones, for the diagnosis of GPA.

Methods: The evaluated assays for PR3-ANCA detection were representative of the first, second, and third generation tests (direct, capture and anchor assays, respectively). A third-generation assay employing both human and recombinant PR3 was also evaluated. The study population consisted of 55 GPA patients, 175 disease controls (representing most diseases in differential diagnosis with primary small-vessel vasculitis) including 52 with microscopic polyangiitis, and 20 healthy subjects. We performed the primary evaluation of test sensitivity using cut-off points which provided adequate and identical specificity for each test.

Results: Although specificity and area under the ROC curve did not differ significantly between the different assays, substantial differences in sensitivity at 98%-specificity were found in some instances (p<0.001). Compared to first generation direct PR3-ANCA specific assays, some of the second and third generation tests increased the positive predictive value (PPV) for GPA diagnosis.

Conclusions: Some of the newer PR3-ANCA specific assays have better PPV than traditional ones.

Keywords: antineutrophil cytoplasmic autoantibodies (ANCA); granulomatosis with polyangiitis (GPA); proteinase 3 (PR3)
Corresponding author: Antonella Radice, Microbiology Institute, San Carlo Borromeo Hospital, Via Pio II 3, 20153 Milan, Italy, Phone: +39 02 40222457, Fax: +39 02 40222829, E-mail:

The authors wish to thank Carlo De Maria and Salvatore Di Marco, with all their staff members, for their continuous technical support and assistance, and the clinical engineering services for the professional help.

Conflict of interest statement

Authors’ conflict of interest disclosure: The authors stated that there are no conflicts of interest regarding the publication of this article.

Research funding: None declared.

Employment or leadership: None declared.

Honorarium: None declared.

References

1. Savige J, Davies D, Falk RJ, Jennette JC, Wiik A. Antineutrophil cytoplasmic antibodies and associated diseases: a review of the clinical and laboratory features. Kidney Int 2000;57:846–62.10.1046/j.1523-1755.2000.057003846.xSearch in Google Scholar

2. Radice A, Sinico RA. Antineutrophil cytoplasmic antibodies (ANCA). Autoimmunity 2005;38:93–103.10.1080/08916930400022673Search in Google Scholar PubMed

3. Sinico RA, Di Toma L, Maggiore U, Bottero P, Radice A, Tosoni C, et al. Prevalence and clinical significance of antineutrophil cytoplasmic antibodies in churg-strauss syndrome. Arthritis Rheum 2005;52:2926–35.10.1002/art.21250Search in Google Scholar PubMed

4. Sinico RA, Radice A, Pozzi C, Ferrario F, Arrigo G. Diagnostic significance and antigen specificity of antineutrophil cytoplasmic antibodies in renal disease. A prospective multicentric study. Nephrol Dial Transplant 1994;9:505–10.10.1093/ndt/9.5.505Search in Google Scholar

5. Boomsma MM, Stegeman CA, van der Leij MJ, Oost W, Hermans J, Kallenberg CG, et al. Prediction of relapses in Wegener’s granulomatosis by measurement of antineutrophil cytoplasmic antibody levels. Arthritis Rheum 2000;43:2025–33.10.1002/1529-0131(200009)43:9<2025::AID-ANR13>3.0.CO;2-OSearch in Google Scholar

6. Kerr GS, Fleisher TA, Hallahan CW, Leavitt RY, Fauci AS, Hoffman GS. Limited prognostic value of changes in antineutrophil cytoplasm antibody titer in patients with Wegener’s granulomatosis. Arthritis Rheum 1993;36:365–71.10.1002/art.1780360312Search in Google Scholar

7. Girard T, Mahr A, Noel LH, Cordier JF, Lesavre P, Andrè MH. Are antineutrophil cytoplasmic antibodies a marker predictive of relapse in Wegener’s granulomatosis? A prospective study. Rheumatology (Oxford) 2001;40:147–51.10.1093/rheumatology/40.2.147Search in Google Scholar

8. Han WK, Choi HK, Roth RM, McCluskey RT, Niles JL. Serial ANCA titers: useful tool for prevention of relapses in ANCA-associated vasculitis. Kidney Int 2003;63:1079–85.10.1046/j.1523-1755.2003.00821.xSearch in Google Scholar PubMed

9. Savige J, Gillis D, Benson E, Davies D, Esnault V, Falk R, et al. International consensus statement on testing and reporting of Antineutrophil Cytoplasmic Antibodies (ANCA). Am J Clin Pathol 1999;111:507–13.10.1093/ajcp/111.4.507Search in Google Scholar PubMed

10. Savige J, Dimech W, Fritzler M, Goeken J, Hagen EC, Jennette JC, et al. For the international group for consensus statement on testing and reporting of Antineutrophil Cytoplasmic Antibodies (ANCA). Addendum to the international consensus statement on testing and reporting of antineutrophil cytoplasmic antibodies. Am J Clin Pathol 2003;120:312–8.10.1309/WAEPADW0K4LPUHFNSearch in Google Scholar

11. Wiik A. Delineation of a standard procedure for indirect immunofluorescence detection of ANCA. APMIS 1989;6:12–3.Search in Google Scholar

12. Radice A, Vecchi M, Bianchi MB, Sinico RA. Contribution of immunofluorescence to the identification and characterization of anti-neutrophil cytoplasmic autoantibodies. The role of different fixatives. Clin Exp Rheumatol 2000;18:707–12.Search in Google Scholar PubMed

13. Goldschmeding R, van der Schoot CE, ten Bokkel Huinink D, Hack CE, van den Ende ME, Kallenberg CG. Wegener’s granulomatosis autoantibodies identify a novel diisopropylfluorophosphate-binding protein in the lysosomes of normal human neutrophils. J Clin Invest 1989;84:1577–87.10.1172/JCI114335Search in Google Scholar

14. Radice A, Sabadini E, Sinico RA. Antineutrophil cytoplasmic autoantibodies with specificity for proteinase 3. In: Shoenfeld Y, Gershwin E, Meroni PL, editors. Autoantibodies. Amsterdam: Elsevier, 2006.10.1016/B978-044452763-9/50018-4Search in Google Scholar

15. Merkel PA, Polisson RP, Chang YC, Skates SJ, Niles JL. Prevalence of antineutrophil cytoplasmic antibodies in a large inception cohort of patients with connective tissue disease. Ann Intern Med 1997;126:866–73.10.7326/0003-4819-126-11-199706010-00003Search in Google Scholar

16. Jennette JC, Wilkman AS, Falk RJ. Diagnostic predictive value of ANCA serology. Kidney Int 1998;53:796–8.10.1038/ki.1998.36Search in Google Scholar PubMed

17. Brenner H, Gefeller O. Variation of sensitivity, specificity, likelihood ratios and predictive values with disease prevalence. Stat Med 1997;16:981–91.10.1002/(SICI)1097-0258(19970515)16:9<981::AID-SIM510>3.0.CO;2-NSearch in Google Scholar PubMed

18. Wang G, Csernok E, de Groot K, Gross WL. Comparison of eight commercial kits for quantitation of antineutrophil cytoplasmic antibodies (ANCA). J Immunol Methods 1997;208:203–11.10.1016/S0022-1759(97)00154-3Search in Google Scholar

19. Csernok E, Ahlquist D, Ullrich S, Gross WL. A critical evaluation of commercial immunoassays for antineutrophil cytoplasmic antibodies directed against proteinase 3 and myeloperoxidase in Wegener’s granulomatosis and microscopic polyangiitis. Rheumatology (Oxford) 2002;41:1313–7.10.1093/rheumatology/41.11.1313Search in Google Scholar

20. Holle JU, Hellmich B, Backes M, Gross WL, Csernok E. Variations in performance characteristics of commercial enzyme immunoassay kits for detection of antineutrophil cytoplasmic antibodies: what is the optimal cut off? Ann Rheum Dis 2005;64:1773–9.10.1136/ard.2005.035279Search in Google Scholar PubMed

21. Sommarin Y, Rasmussen N, Wieslander J. Characterization of monoclonal antibodies to proteinase 3 and application in the study of epitopes for classical anti-neutrophil cytoplasm antibodies. Exp Nephrol 1995;3:249–56.Search in Google Scholar

22. Erdbrugger U, Hellmark T, Bunch DO, Alcorta DA, Jennette JC, Falk RJ, et al. Mapping of myeloperoxidase epitopes recognized by MPO-ANCA using human-mouse MPO chimers. Kidney Int 2006;69:1799–805.10.1038/sj.ki.5000354Search in Google Scholar PubMed

23. Westman KW, Selga D, Bygren P, Segelmark M, Baslund B, Wiik A, et al. Clinical evaluation of a capture ELISA for detection of proteinase 3 antineutrophil cytoplasmic antibody. Kidney Int 1998;53:1230–6.10.1046/j.1523-1755.1998.00873.xSearch in Google Scholar PubMed

24. Csernok E, Holle J, Hellmich B, Willem J, Tervaert C, Kallenberg CG, et al. Evaluation of capture ELISA for detection of antineutrophil cytoplasmic antibodies directed against proteinase 3 in Wegener’s granulomatosis: first results from a multicentre study. Rheumatology (Oxford) 2004;43:174–80.10.1093/rheumatology/keh028Search in Google Scholar

25. Hellmich B, Csernok E, Fredenhagen G, Gross WL. A novel high sensitivity ELISA for detection of antineutrophil cytoplasm antibodies against proteinase-3. Clin Exp Rheumatol 2007;25:1–5.Search in Google Scholar

26. Radice A, Bianchi L, Palumbo A, Sinico RA. Clinical evaluation of the new AeskulisaPR3sensitiv assay for the diagnosis of Wegener granulomatosis. APMIS 2009;117(s127):180.Search in Google Scholar

27. Damoiseaux J, Dähnrich C, Rosemann A, Probst C, Komorowski L, Stegeman CA, et al. A novel enzyme-linked immunosorbent assay using a mixture of human native and recombinant proteinase-3 significantly improves the diagnostic potential for antineutrophil cytoplasmic antibody-associated vasculitis. Ann Rheum Dis 2009;68:228–33.http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000262394000013&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f310.1136/ard.2007.086579Search in Google Scholar

28. Radice A, Bianchi L, Palumbo A, Sinico RA. A novel PR3-ANCA ELISA with a mixture of human purified and recombinant PR3 as solid phase antigen. In: Conrad K, Chan EK, Fritzler MJ, Humbel RL, von Landenberg P, Shoenfeld Y, editors. From pathogenesis to therapy of autoimmune diseases. PABST Science Publishers 2009;6:383–5.Search in Google Scholar

29. Fries JF, Hunder GG, Bloch DA, Michel BA, Arend WP, Calabrese LH, et al. The American College of Rheumatology, criteria for the classification of vasculitis. Summary. Arthritis Rheum 1990;33:1135–6.http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000283059100003&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar PubMed

30. Jennette JC, Falk RJ, Andrassy K, Bacon PA, Churg J, Gross WL, et al. Nomenclature of systemic vasculitides. Proposal of an international consensus conference. Arthritis Rheum 1994;37:187–92.10.1002/art.1780370206http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000312938100001&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar PubMed

31. Watts R, Lane S, Hanslik T, Hauser T, Hellmich B, Koldingsnes W, et al. Development and validation of a consensus methodology for the classification of the ANCA-associated vasculitides and polyarteritis nodosa for epidemiological studies. Ann Rheum Dis 2007;66:222–7.http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000243639500015&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f310.1136/ard.2006.054593Search in Google Scholar PubMed

32. Ruperto N. For the Paediatric Rheumatology International Trials Organisation (PRINTO). Classification criteria and diagnostic tests for vasculitides. J Rheumatol 2012;39:1503–5.10.3899/jrheum.120640Search in Google Scholar

33. Sinico RA, Pozzi C, Radice A, Tincani A, Li Vecchi M, Rota S, et al. Clinical significance of antineutrophil cytoplasmic autoantibodies with specificity for lactoferrin in renal diseases. Am J Kidney Dis 1993;22:253–60.10.1016/S0272-6386(12)70314-1Search in Google Scholar PubMed

34. Obuchowski N, McClish D. Sample size determination for diagnostic accuracy studies involving binormal ROC curve indices. Stat Med 1997;16:1529–42.10.1002/(SICI)1097-0258(19970715)16:13<1529::AID-SIM565>3.0.CO;2-HSearch in Google Scholar PubMed

35. Li J, Fine J. On sample size for sensitivity and specificity in prospective diagnostic accuracy studies. Stat Med 2004;23:2537–50.10.1002/sim.1836Search in Google Scholar PubMed

36. Stata Base Reference Manual Release 11, Vol. 3. Stata Press Publication, StataCorp, College Station, TX, USA, 2009: 1595–615.Search in Google Scholar

37. DeLong ER, DeLong DM, Clarke-Pearson DL. Comparing the areas under two or more correlated receiver operating characteristics curve: a non parametric approach. Biometrics 1988;44:837–45.10.2307/2531595Search in Google Scholar

38. Seed PT, Tobias A. Summary statistics for diagnostic tests. Stata J 2004;4:490.10.1177/1536867X0400400414Search in Google Scholar

39. Hawass NE. Comparing the sensitivities and specificities of two diagnostic procedures performed on the same group of patients. Br J Radiol 1997;70:360–6.10.1259/bjr.70.832.9166071Search in Google Scholar PubMed

40. Holle JU, Herrmann K, Gross WL, Csernok E. Comparative analysis of different commercial ELISA systems for the detection of anti-neutrophil cytoplasm antibodies in ANCA-associated vasculitides. Clin Exp Rheumatol 2012;30(Suppl 70):S66–9.Search in Google Scholar

41. Hagen EC, Daha MR, Hermans J, Andrassy K, Csernok E, Gaskin G, et al. Diagnostic value of standardized assays for anti-neutrophil cytoplasmic antibodies in idiopathic systemic vasculitis. EC/BCR Project for ANCA Assay Standardization. Kidney Int 1998;53:743–53.10.1046/j.1523-1755.1998.00807.xSearch in Google Scholar
Received: 2013-04-23
Accepted: 2013-05-27
Published Online: 2013-06-25
Published in Print: 2013-11-01

©2013 by Walter de Gruyter Berlin Boston

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https://doi.org/10.1515/cclm-2013-0308
Keywords for this article
antineutrophil cytoplasmic autoantibodies (ANCA); granulomatosis with polyangiitis (GPA); proteinase 3 (PR3)

Articles in the same Issue

  1. Letter to the Editor
  2. Elevated level of cell-free plasma DNA is associated with advanced-stage breast cancer and metastasis
  3. Is procalcitonin a reliable marker of sepsis in critically ill septic patients undergoing continuous veno-venous hemodiafiltration with “high cut-off” membranes (HCO-CVVHDF)?
  4. Challenging our serological algorithm for celiac disease (CD) diagnosis by the ESPGHAN guidelines
  5. Preanalytical quality control in a university hospital in China
  6. Racial differences and relationships between gestational thyrotropin and free thyroxine in a multiracial Asian population
  7. Analytical performance and method comparison study of the total homocysteine immunoassay on the AIA 600II analyser
  8. Easy verification of clinical chemistry reference intervals
  9. The Elecsys® Vitamin B12 assay is not affected by anti-intrinsic factor antibodies
  10. Chemiluminescence-based cobalamin assay errors: background and perspectives
  11. Congress Abstracts
  12. Abstracts IV Italian Great Network Congress Rome, 14th–18th October 2013*)
  13. Masthead
  14. Masthead
  15. Editorial
  16. Making colorectal cancer screening FITTER for purpose with quantitative faecal immunochemical tests for haemoglobin (FIT)
  17. From “panic” to “critical” values: which path toward harmonization?
  18. Review
  19. Cerebrospinal fluid analyses for the diagnosis of subarachnoid haemorrhage and experience from a Swedish study. What method is preferable when diagnosing a subarachnoid haemorrhage?
  20. Opinion Paper
  21. False myths and legends in laboratory diagnostics
  22. General Clinical Chemistry and Laboratory Medicine
  23. National survey on critical values notification of 599 institutions in China
  24. Influence of physical properties of cuvette surface on measurement of serum lipase
  25. Red cell indices: differentiation between β-thalassemia trait and iron deficiency anemia and application to sickle-cell disease and sickle-cell thalassemia
  26. Measurement of immature platelets with Abbott CD-Sapphire and Sysmex XE-5000 in haematology and oncology patients
  27. Performance characteristics of consensus approaches for small and minor paroxysmal nocturnal hemoglobinuria clone determination by flow cytometry
  28. Comparison of PR3-ANCA specific assay performance for the diagnosis of granulomatosis with polyangiitis (Wegener’s)
  29. The integration of the detection of systemic sclerosis-associated antibodies in a routine laboratory setting: comparison of different strategies
  30. Reference Values and Biological Variations
  31. Reference interval studies: what is the maximum number of samples recommended?
  32. Short-term estimation and application of biological variation of small dense low-density lipoproteins in healthy individuals
  33. Cancer Diagnostics
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