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ANCA-C (PR3) ELISA Kit

INTENDED USE

Human ANCA-C ELISA kit is designed for measuring IgG autoantibodies to proteinase 3 (PR3, anti-PR3, ANCA-C, cytoplasmic anti-neutrophil cytoplasmic antibodies, c-ANCA) using human serum or plasma. This assay has a minimum analytical sensitivity limit of 0.5 U/ml.

BACKGROUND

Cytoplasmic anti-neutrophil cytoplasmic antibodies (cANCA, ANCA-C or ANCA-PR3) are autoantibodies which are produced by the body to attack its own proteins. Proteinase 3 (PR3) is one of the most common target protein and it can occur throughout the neutrophil cytoplasm. It is predominately found in granulomatosis with polyangiitis patients (over 90%). PR3 is also known to catabolise a variety of human proteins for example elastin, four subclasses of IgG (including ANCA-PR3 and PR3 complex) and proteoglycans. The following ELISA method can be used to interpret samples that are difficult to analyse by indirect immunofluorescence techniques such as those with high background fluorescence or those which exhibit several antibodies simultaneously. It is widely accepted that ANCA-C, cANCA and ANCA-PR3 is present in around 80% of patients with Wegener’s granulomatosis and almost 35% of patients with microscopic polyangiitis and Churg-Strauss syndrome.

ANCA-C (PR3) ELISA KIT CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

  • Divisible Microplate: Highly purified proteinase 3 (PR3) is bound to microwells.
  • Calibrator A-F: Concentration 0, 5, 10, 20, 40, 100 U/ml.
  • Control Positive And Control Negative: Human PR3 antibodies in a serum/buffer matrix.
  • Sample Buffer (5x).
  • Enzyme Conjugate: Containing anti-human IgG antibodies, HRP labelled.
  • Substrate: Tetramethylbenzidin (TMB).
  • Stop Solution.
  • Wash Buffer (50x).
  • Instruction For Use.
  • Certificate Of Analysis.

SENSITIVITY

The minimum detection sensitivity level of human IgG autoantibodies to proteinase 3 (anti-PR3, PR3, ANCA-C, c-ANCA, cytoplasmic anti-neutrophil cytoplasmic antibodies) using current human ANCA-C ELISA kit was 0.5 U/ml. The dynamic range for this assay is 5 – 100.0 U/ml.

ASSAY CHARACTERISTICS

– ANCA-C (PR3): ELISA
– Measuring Range: 0 – 100 U/ml
– Expected Values: In a normal range = Cut-off Index 5.0 U/ml, Positive: Index ≥ 5.0 U/ml, Negative: Index < 5.0 U/ml.
– Linearity: 96 – 108%
– Intra Assay Precision: 2.2 – 4.7%
– Inter Assay Precision: 3.9 – 8.8%
– Clinical Diagnosis: Sensitivity (85.2%), Specificity (98.8%), Overall agreement (96.1%).

REFERENCES

  1. Human proteinase 3 (PR3) and its binding molecules: implications for inflammatory and PR3-related autoimmune responses. Ann N Y Acad Sci. (2007) 1109: 84-92. Müller A., et al.
  2. PR3-ANCA: a promising biomarker for ulcerative colitis with extensive disease. Clin Chim Acta. (2013) 424: 267-73. Mahler M., et al.
  3. Antibodies directed against neutrophils (C-ANCA and P-ANCA) are of distinct diagnostic value in systemic vasculitis. Q J Med. (1990) 77 (284): 1287-96. Venning M.C., et al.
  4. Performance evaluation of three assays for the detection of PR3-ANCA in granulomatosis with polyangiitis in daily practice. Autoimmun Rev. (2013) 12 (12): 1118-22. Noel N, André C., et al.

ADDITIONAL INFORMATION

  • Full Name: ANCA-C (PR3) ELISA Kit
  • Reactivity: Human
  • Sample Type: Serum, Plasma
  • Sensitivity: 0.5 U/ml

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