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ANCA Screen ELISA Kit

INTENDED USE

Human ANCA Screen ELISA kit is a method intended for screening in vitro quantitative determination of IgG class autoantibodies directed against PR3 and MPO (ANCA screen) in human serum and plasma samples. This assay has a minimum analytical sensitivity limit of diagn. 95.1%.

A mixture of purified antigens PR3 and MPO is coated on to microwells.

BACKGROUND

Anti-neutrophil cytoplasmic antibodies (ANCA) are mostly IgG type group of autoantibodies directed towards the cytoplasmic components of neutrophil granulocytes (majority are white blood cells) and monocytes. Often found in many different autoimmune diseases associated to systemic vasculitis that can be measured using blood tests. Examples of some of the common disorders include eosinophilic granulomatosis with polyangiitis, granulomatosis with polyangiitis, drug induced vasculitides and microscopic polyangiitis.

Two method used frequently to measure ANCA are ELISA and indirect immunofluorescence. Using the ELISA assay it is possible to analyse specific antibodies to a particular antigen, once a positive screen has been carried out. MPO and PR3 are two of the most measured antigens.

ANCA SCREEN 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: Anti-neutrophil cytoplasmic antibodies (MPO and PR3) are bound to microwells.
  • Control A (Negative), Control B (Cut-Off), Control C (Positive).
  • Sample Buffer (5x Conc.).
  • Enzyme Conjugate.
  • Substrate: Tetramethylbenzidin (TMB).
  • Stop Solution.
  • Wash Buffer (50x).
  • Instruction For Use.
  • Certificate Of Analysis.

SENSITIVITY

The minimum detection sensitivity level of human class autoantibodies directed against PR3 or MPO using current ANCA screen ELISA kit was diagn. 95.1%.The dynamic range for this assay is cut-off index 1.0.

ASSAY CHARACTERISTICS

– Anti-Neutrophil Cytoplasmic Antibodies (ANCA Screen): ELISA
– Expected Values: In a normal range = Cut-off Index 1.0, Positive: Index ≥ 1.0, Negative: Index < 1.0.
– Intra Assay Precision: 3.3 – 5.4%
– Inter Assay Precision: 4.5 – 11.9%
– Clinical Diagnosis: Sensitivity (95.1%), Specificity (97.7%), Overall agreement (96.5%).

REFERENCES

  1. Incidence of PR3- and MPO-ANCA autoantibody specificity changes in ANCA-associated vasculitis. Ann Clin Biochem. (2015) 52 (Pt 2): 297-301. Holding S., et al.
  2. How are antineutrophil cytoplasmic autoantibodies detected? Am J Kidney Dis. (1991) 18 (2): 154-8. Wieslander J.
  3. What do antineutrophil cytoplasmic antibodies (ANCA) tell us? Best Pract Res Clin Rheumatol. (2005) 19 (2): 263-76. Review. Savige J., et al.
  4. Audit of the clinical usefulness of a rapid qualitative ELISA screen for antimyeloperoxidase and antiproteinase 3 antibodies in the assessment of patients with suspected vasculitis. J Clin Pathol. (2003) 56 (10): 775-7. Aslam A., et al.
  5. Autoantibody detection using multiplex technologies. Lupus. (2006) 15 (7): 412-21. Review. Binder S.R.

ADDITIONAL INFORMATION

  • Full Name: ANCA Screen ELISA Kit
  • Reactivity: Human
  • Sample Type: Serum, Plasma
  • Sensitivity: 95.1%

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