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ANCA-P (MPO) ELISA Kit

INTENDED USE

Human ANCA-P ELISA kit is a procedure designed for detecting IgG autoantibodies to myeloperoxidase (MPO, anti-MPO, ANCA-P, perinuclear anti-neutrophil cytoplasmic antibodies, p-ANCA) using human plasma and serum. This assay has a minimum analytical sensitivity limit of 0.5 U/ml.

BACKGROUND

Perinuclear anti-neutrophil cytoplasmic antibodies (p-ANCA, MPO-ANCA, anti-MPO, ANCA-P) display a perinuclear staining pattern and are known to be common targets for myeloperoxidase (MPO). MPO is a neutrophil granule protein which has a primary function during normal metabolic processes which leads to the generation oxygen radicals. However, the amount of pANCA sera that displays anti-MPO antibodies is only reported in very low number of patients having vasculitis condition, this is a little as even 12%.

MPO is described as a cationic protein that is predominately found in azurophilic granules that are present in monocytes or neutrophils. It is also found that MPO can utilise chloride ions and H2O2 in order top form hydochlorous (HOCl), this is known to possess a potent cytotoxic oxidant activity. There is evidence to indicate that anti-MPO antibodies can play pathogenic functions during the process of vasculitides necrotising both in vivo and in vitro. Perinuclear anti-neutrophil cytoplasmic antibodies are associated with many different medical conditions such as rheumatoid arthritis, primary sclerosing cholangitis, ulcerative colitis, eosinophilic granulomatosis with polyangiitis, microscopic polyangiitis, crescentic glomerulonephritis and focal necrotising.

The analysis of anti-PR3 and anti-MPO levels has been found to correlate with the clinical status and both display elevated levels in active diseases and the antibody titres start to decrease during therapy, eventually becoming undetectable after remission. The most common protocol that is used in the detection of ANCA are either ELISA assay or immunofluorescence tests. Both methods will help to identify the two main patterns that are either cytoplasmic (cANCA) and perinuclear (pANCA) type.

ANCA-P (MPO) 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 myeloperoxidase (MPO) is bound to microwells.
  • Calibrator A-F: Concentration ranging between 0 – 100 U/ml.
  • Control Positive and Control Negative: Human MPO antibodies in a serum/buffer matrix.
  • Sample Buffer (5x).
  • Enzyme Conjugate: Anti-human IgG antibodies, HRP labelled.
  • TMB Substrate.
  • Stop Solution.
  • Wash Buffer (50x).
  • Instruction for Use.
  • Certificate of Analysis.

SENSITIVITY

The minimum detection sensitivity level of human IgG autoantibodies to myeloperoxidase (MPO, anti-MPO, P-ANCA, p-ANCA, perinuclear anti-neutrophil cytoplasmic antibodies, ) using current ANCA-P ELISA kit was 0.5 U/ml. The dynamic range for this assay is 5 – 100.0 U/ml.

ASSAY CHARACTERISTICS

– ANCA-P (MPO): ELISA
– Measuring Range: 0 – 100 U/ml
– Expected Values: In a normal range = Cut-off Index 5.0 U/ml, Negative: Index < 5.0 U/ml, Positive: Index ≥ 5.0 U/ml.
– Linearity: 85 – 101%
– Intra Assay Precision: 3.1 – 6.4%
– Inter Assay Precision: 4.9 – 6.3%
– Clinical Diagnosis: Sensitivity (98.2%), Specificity (96.7%), Overall agreement (97.1%).

REFERENCES

  1. Comparison of a novel chemiluminescence enzyme immunoassay (CLEIA) with enzyme-linked immunosorbent assay (ELISA) for the determination of MPO-ANCA in patients with ANCA-associated vasculitis. Mod Rheumatol. (2015) 25 (2): 230-4. Hirose O., et al.
  2. Pulmonary hyalinizing granuloma associated with Sjögren syndrome and ANCA MPO vasculitis. Joint Bone Spine. (2015) 82 (1): 71-2. Rodríguez-Muguruza S., et al.
  3. Myeloperoxidase (MPO)-specific CD4+ T cells contribute to MPO-anti-neutrophil cytoplasmic antibody (ANCA) associated glomerulonephritis. Cell Immunol. (2013) 282 (1): 21-7. Gan P.Y., et al.
  4. Getting rid of MPO-ANCA: a matter of disease subtype. Rheumatology (Oxford). (2013) 52 (4): 752-4. Bremer J.P., et al.
  5. What is myeloperoxidase doing in ANCA-associated glomerulonephritis? Kidney Int. (2015) 88 (5): 938-40. Couser W.G. and Johnson R.J.

ADDITIONAL INFORMATION

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

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