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ANCA Profile ELISA Kit (Anti-Neutrophil Cytoplasmic Antibodies)

INTENDED USE

Human ANCA profile ELISA kit is intended for profiling in vitro quantitative levels of IgG anti-neutrophil cytoplasmic antibodies (ANCA-IgG, ANCA profile) in human plasma and serum. This assay has a minimum analytical sensitivity limit of diagn. 90.2%.

Purified antigens PR3, MPO, BPI, Elastase, Cathepsin G, Lysozyme and Lactoferrin are coated on to individual rows A to H of the microwell plate.

BACKGROUND

Anti-neutrophil cytoplasmic antibodies (ANCA) is an IgG type autoantibodies that is directed towards the cytoplasmic components of monocytes and neutrophil granulocytes. They are analysed by carry out various blood tests to identify many different autoimmune disorders, especially systemic vasculitis (ANCA-associated vasculitides). They are associated with many types of vasculitides for example: microscopic polyangiitis, drug induced vasculitides, polyangiitis and eosinophilic granulomatosis.

There are many small blood vessels present within the human body, therefore this increases the likelihood of affecting any part of the body. Some of the common affected body parts include: joints, kidneys, ears, lungs, nerves and nose. Due to the lungs and the kidneys representing essential organs, it is important to identify ANCA-associated vasculitis at an early stage since this prevent these organs from being seriously damaged in the future.

During the process of ANCA-associated vasculitis, it is found that ANCA can specially interact and bind to two proteins that are present in cytoplasm (mainly the fluids in neutrophils). They are known to have a major contribution to causing damage to blood vessels. The actual process and the specific mechanism how it becomes present in different patients especially at different age group is still not very clear and researchers are still working hard to understand this better.

ELISA assays procedure has been effectively used in diagnostic research institutes even though indirect immunofluorescence (IF) is favoured for screening. One of the major advantages of using the ELISA method is that it can detect specific antibodies to an individual antigen, some of the most common antigens that are used include PR3 and MPO (usually tested after a positive IF test).

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 are bound to microwells.
  • Control A (Negative), Control B (Cut-Off), Control C (Positive).
  • Sample Buffer (5x).
  • Enzyme Conjugate.
  • TMB Substrate.
  • Stop Solution.
  • Wash Buffer (50x Conc.).
  • Instruction For Use.
  • Certificate Of Analysis.

SENSITIVITY

The minimum detection sensitivity level of IgG autoantibodies to human neutrophil cytoplasmic antigens (ANCA-IgG) using current ANCA ELISA kit was diagn. 90.2%.The dynamic range for this assay is cut-off index 1.0.

ASSAY CHARACTERISTICS

– Expected Values: In a normal range = Cut-off Index 1.0, Negative: Index < 1.0 Positive: Index ≥ 1.0.
– Linearity: 83 – 106%
– Intra Assay Precision: 2.8 – 5.1%
– Inter Assay Precision: 2.0 – 3.7%
– Clinical Diagnosis: Sensitivity (90.2%), Specificity (86.3%), Overall agreement (87.5%).

REFERENCES

  1. Expression profile of leukocyte genes activated by anti-neutrophil cytoplasmic autoantibodies (ANCA). Kidney Int. (2002) 62 (5): 1638-49. Yang J.J., et al.
  2. A novel autoantibody against moesin in the serum of patients with MPO-ANCA-associated vasculitis. Nephrol Dial Transplant. (2014) 29 (6): 1168-77. Suzuki K., et al.
  3. Clinical use of serological tests for antineutrophil cytoplasmic antibodies. What do the studies say? Rheum Dis Clin North Am. (2001) 27 (4): 799-813, vi-vii. Review. Wiik AS.
  4. Implication of B lymphocytes in the pathogenesis of ANCA-associated vasculitides. Autoimmun Rev. (2015) 14 (11): 996-1004. Review. Dumoitier N., et al.
  5. Expression profile of proinflammatory genes in neutrophil-enriched granulocytes stimulated with native anti-PR3 autoantibodies. J Physiol Pharmacol. (2012) 63 (3): 249-56. Surmiak M., et al.

ADDITIONAL INFORMATION

  • Full Name: ANCA Profile ELISA Kit (Anti-Neutrophil Cytoplasmic Antibodies)
  • Reactivity: Human
  • Sample Type: Plasma, Serum
  • Sensitivity: 90.2%

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