Vasculitis EIA

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Overview

Vasculitis EIA

Anti-PR3 (c-ANCA) Autoimmune EIA

Clinical Utility

The systemic vasculitides are inflammatory diseases of blood vessels and comprise a heterogeneous group of disorders, the causes of which are generally unknown. The diseases have diverse presentations, and are often rapidly progressive, causing irreversible damage to kidney and lung blood vessels. Davies1 first observed antineutrophil cytoplasmic antibodies (ANCA) in vasculitis patients in 1982. ANCA are autoantibodies specific for proteins located in the primary and secondary granules of neutrophils and in the peroxidase-positive lysosomes of peripheral blood monocytes. They were originally detected by indirect immunofluorescence on ethanol-fixed neutrophils, producing characteristic staining patterns with accentuation of the fluorescent activity within the nuclear lobes. Two major patterns of immunofluorescent staining have been observed: a classical or cytoplasmic staining designated c-ANCA, and a perinuclear pattern designated p-ANCA, which was described in association with renal systemic vasculitis.2 Other staining patterns have been described and are generally noted as atypical or snowdrift patterns.3,4 The nature of the antigens and clinical significance of the antibodies responsible for these atypical fluorescent patterns is currently unclear.

The association between ANCA and Granulomatosis with polyangitis (Wegener's/GPA), a systemic necrotising vasculitis was formally described by Woude in 1985, and confirmed by other workers.5,6 The target autoantigen associated with c-ANCA was identified in 1990 as proteinase 3,7 a 29 kD neutrophil serine protease of azurophil granules, previously characterized by Kao.8 Proteinase 3 has been confirmed as the major autoantigen in GPA.9,10

The presence of c-ANCA denotes a spectrum of disease varying from idiopathic pauci-immune necrotizing glomerulonephritis to GPA. GPA is characterized by granulomatomous inflammation of the respiratory tract, systemic vasculitis and necrotising crescentric glomerulonephritis; c-ANCA are present in over 90% of GPA patients.11–13 It is also detected in 67–86% of patients with so-called limited GPA without renal involvement, and in 40–50% of patients with pauci-immune necrotizing glomerulonephritis.

Not all c-ANCA-positive sera react with the PR3 antigen; reported sensitivity of anti-PR3 antibodies is 70–100%.14–16 c-ANCA-positive serum may contain antibodies against antigens other than PR3 but the role of these antibodies is unclear. Prognostically, antibody titer rises are thought to predict relapse and to help differentiate relapse from opportunistic infection.17,18 Recent evidence suggests that in a large minority of patients, c-ANCA/anti-PR3 titers do not follow disease activity.19

As the clinical spectrum of ANCA-related diseases increases and further target antigens are identified and characterized, the introduction of antigen-specific ELISAs using highly purified antigen extracts may play a valuable role in the identification of disease subtypes.

References

  1. Davies DJ, et al. Br Med J. 285:606, 1982.
  2. Falk RJ, et al. N Engl J Med. 318:1651–1657, 1988.
  3. Rump JA, et al. Immunobiology. 181:406–413, 1990.
  4. Hagen EC, et al. Blood. 81:1996–2002, 1993.
  5. Gross WL, et al. Lancet. 5:806, 1986.
  6. Parlevliet KJ, et al. Q J Med. 66:55–63, 1988.
  7. Ludemann J, et al. J Exp Med. 171:357–362, 1990.
  8. Kao RC, et al. J Clin Invest. 82:1963–1973, 1988.
  9. Jennette JC, et al. Blood. 75:2263–2264, 1990.
  10. Lesavre P, et al. Klin Wochenschr. 69:552–557, 1991.
  11. Cohen Tervaert JW, et al. Arch Int Med. 149:2461–2465, 1989
  12. Andrassy K, et al. Clin Nephrol. 32:159–167, 1989.
  13. Gross WH, et al. Clin Exp Immunol. 91:1–12, 1993.
  14. Kallenberg CG, et al. Amer J Med. 93:675–682, 1992.
  15. Falk RJ and Jennette JC. Am J Kidney Dis. 18:145–147, 1991.
  16. Gross WL, et al. 5th International ANCA Workshop,Cambridge, UK, September 1993.
  17. Nolle B, et al. Ann Intern Med. 111 28–40, 1989.
  18. Cohen Tervaert JW, et al. Lancet. 336:709–711, 1990.
  19. Kerr GS, et al. Arthritis Rheum36:365–371, 1993.
Vasculitis EIA

Anti-PR3 Autoimmune EIA

Result Interpretation

Qualitative
Absorbance Ratio

Result Interpretation
OD Patient ≤ OD Cutoff Negative
OD Patient > OD Cutoff Positive
Semi-Quantitative
< 15 U/mL Negative
> 15 U/mL
Positive

Bio-Rad offers three autoimmune EIA tests for vasculitis: Anti-PR3, Anti-MPO and Anti-GBM. The Bio-Rad Autoimmune Vasculitis EIA product line features common assay protocols, common reagents, color-coded breakaway wells and easy automation, allowing for simple, objective and sensitive diagnostics.

Assay Characteristics

  • Sample dilution: 1:101
  • Sample volume: 10 µL
  • Incubation time: 90 minutes (30/30/30)
  • Incubation temperature: room temperature (18–25°C)
  • Conjugate: Horseradish Peroxidase
  • Substrate: Tetramethylbenzidine (TMB) and hydrogen peroxide (H2O2)

The Bio-Rad Autoimmune EIA Anti-Proteinase 3 (anti-PR3) test is a solid phase enzyme immunoassay employing highly purified human proteinase 3 (PR3) from human neutrophil granulocytes for the semi-quantitative and qualitative EIA for the detection of antibodies against proteinase 3 antigen in human serum. This assay is an aid in the diagnosis of ANCA-associated vasculitis (AAV) including Granulomatosis with polyangiitis (Wegener's) (GPA) and should be used in conjuction with other laboratory and clinical findings.

The Bio-Rad Autoimmune EIA Anti-Myeloperoxidase (anti-MPO) Test is a solid phase enzyme immunoassay employing purified native myelooeroxidase (MPO) from human peripheral blood polymorohonuclear cells for the semi-quantitative and qualitative detection of autoantibodies against myeloperoxidase antigen in human serum or plasma. This assay is an aid in the diagnosis of ANCA-associated vasculitis (AAV), such as polyangitis and glomerulonephritis, and should be used in conjunction with other laboratory and clinical findings.

Bio-Rad Autoimmune Anti-PR3 EIA Kit
Bio-Rad Autoimmune Anti-PR3 EIA Kit

425-2420
Semi-quantitative enzyme immunoassay for detection of autoantibodies specific for proteinase-3 (PR3) antigen (96 tests)

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Bio-Rad Autoimmune Anti-MPO EIA Kit
Bio-Rad Autoimmune Anti-MPO EIA Kit

425-2380
Semi-quantitative enzyme immunoassay for detection of autoantibodies specific for myeloperoxidase (MPO) antigen (96 tests)

List Price:   Inquire
Quantity:   Add to Quote
Add to Hot List