Autoantibodies against Granulocyte Cytoplasm (cANCA/pANCA)

Indirect Immunofluorescence Test: EUROPLUS™ Granulocyte Mosaic 25

: EUROPLUS™ Granulocyte Mosaic 25 (granuloc. (EOH), granuloc. (HCHO), PR3, MPO, GBM, HEp-2 + granuloc. (EOH)): pANCA.

Screening test for the detection of antibodies against granulocyte cytoplasm (ANCA).
Indications: Wegener‘s granulomatosis, various forms of glomerulonephritis, primary sclerosing cholangitis, ulcerative colitis, Crohn's disease.
Initial dilution: serum undiluted and 1 : 10 in parallel; conjugate class anti-human IgG, FITC-labelled.
Using ethanol-fixed granulocytes, antibodies against granulocyte cytoplasm can be detected. In this case, two fluorescence patterns have to be differentiated: a granular fluorescence which is distributed evenly over the entire cytoplasm, leaving the cell nuclei free (cytoplasmic type, cANCA) or a smooth fluorescence wrapped ribbon-like around the cell nuclei (perinuclear type, pANCA).
Antibodies against all relevant granulocyte antigens as well as against further, as yet unknown antigens are detected simultaneously:

Pattern	Target antigen	Associated diseases
cANCA	Proteinase 3	Wegener‘s granulomatosis
pANCA	Myeloperoxidase	Microscopic arteritis, Churg-Strauss syndrome, polyarteritis nodosa
pANCA	Elastase	Ulcerative colitis, Crohn's disease, primary sclerosing cholangitis, systemic lupus erythematosus
pANCA	Kathepsin G	Ulcerative colitis, primary sclerosing cholangitis, Crohn's disease
pANCA	Lysozym Colitis	Ulcerative colitis, primary sclerosing cholangitis, Crohn's disease
pANCA	Laktoferrin	Ulcerative colitis, primary sclerosing cholangitis, Crohn's disease, systemic lupus erythematosus, rheumatoid arthritis
pANCA o. cANCA	BPI	Primary sclerosing cholangitis, ulcerative colitis, or pANCA Crohn's disease
pANCA	unknown	Ulcerative colitis, Crohn's disease

The composite substrate HEp-2 cells + granulocytes enables one to differentiate between pANCA and anti-nuclear antibodies (ANA) which can easily be confused when using ethanol-fixed granulocytes: In the case of a positive ANA result all nuclei of the HEp-2 cells fluoresce, whereas in the case of pANCA (as well as cANCA) only the granulocytes fluoresce.
The EUROPLUS™ substrates PR3 and MPO as monospecific tests can confirm results from conventional granulocyte screening tests. Recombinant GBM EUROPLUS™ substrate also provides additional reliability for diagnosis. When fluorescence patters are unclear (e.g. unspecific fluorescence caused by other cytoplasmic antibodies) these substrates facilitate evaluation.

Determination of autoantibodies against neurtrophil antigens and glomerular basement membrane antigens for the serological diagnosis of Wegener's granulomatosis, microscopic polyangiitis, rapid progressive glomerulonephritis, Goodpasture syndrome and other forms of vasculitis.
Membrane chips printed with individual lines of purified and biochemically characterised antigens. Each antigen is coated onto a separate membrance chip, providing optimal efficiency of antibody detection for each protein.
Serum dilution 1 : 100; conjugate class anti-human IgG, alkaline phosphatase-labelled.
The EUROLINE Profile allows the determination of 3 autoantibodies: antibodies against MPO, PR3 and GBM.
The test strips can be automatically incubated and evaluated using the EUROBlotMaster system and the EUROLineScan programme.

: Incubated ELISA ANCA Profile (antigens: proteinase 3, lactoferrin, myeloperoxidase, elastase, cathepsin G, BPI).

Differentiation of antibodies against granulocyte cytoplasm (ANCA).
Indications: Wegener‘s granulomatosis, various forms of glomerulonephritis, primary sclerosing cholangitis, ulcerative colitis, Crohn's disease.
Serum dilution 1 : 100; conjugate class anti-human IgG, POD-labelled.
6 relevant anti-granulocyte antibodies can be detected simultaneously and monospecifically: autoantibodies against proteinase 3, lactoferrin, myeloperoxidase, elastase, cathepsin G, BPI.
1-point calibration, semi-quantitative. Calibrator pool and serum sample on the same microplate strip.
Native antigens, purified by affinity chromatography.
Available individual ELISA (3-point calibration, quantitative):

	Antigen		Order No.
	Proteinase 3 (PR3-hn-hr: native/recombinant)		EA 1201-9601-2 G
	Myeloperoxidase		EA 1211-9601 G