CytoBead® Technology ANCA, ANA, ANA 2, CeliAK and RPGN

The CytoBead® ANCA, ANA, ANA 2, CeliAK and RPGN assays are used for the diagnosis of systemic and organ specific autoimmune diseases, via indirect immunofluorescence.

The kits CytoBead® ANCA, ANA, ANA 2, CeliAK and RPGN consist of reagents to determine IgG autoantibodies against neutrophil cytoplasmic antigens (CytoBead® ANCA and CytoBead® RPGN), nuclear and cytoplasmic antigens (CytoBead® ANA and CytoBead® ANA 2) and IgA or IgG antibodies against endomysium, transglutaminase 2 and deaminated gliadin (CytoBead® CeliAK).

The unique CytoBead® assays combine sensitive screening via native substrates (tissue sections or cells) as well as confirmatory testing via solid phase assays (microparticles) in one reaction

  • Using the well-known method of indirect immunofluorescence, processing of CytoBead® assays is fast and reliable
  • The use of standard glass slides guarantees simple handling
  • The evaluation of CytoBead® assays can be done manually or automatically using the AKLIDES® system (for more information see www.medipan.de)
  • With automatic evaluation using the AKLIDES® system, quantitative results comparable to those of ELISAs are produced.
Video presentation:



All CytoBead® assays utilize indirect immunofluorescence as a basic principle. Using a native substrate (tissue sections or cells) as well as a multiplex solid phase test (beads), IgG and IgA autoantibodies from human serum can be detected in one reaction. In the first step, autoantibodies in the diluted human serum react with the substrates on the cells and tissues, as well as with the appropriate autoantigens on the fluorescently coded beads.

In the second step the specific detection of the antigen-antibody reaction takes place via fluorophore labelled secondary antibodies.

Assay principle

Indirect immunofluorescence

Assay size

48 (6x8) determinations for CytoBead® ANCA, CeliAK and RPGN as well as 80 (10x8) determinations for CytoBead® ANA or CytoBead® ANA2

Parameter

Human granulocytes (ethanol fixated) and microparticles coated with PR3, MPO and GBM (CytoBead® ANCA)

Human granulocytes (ethanol fixated) and microparticles coated with PR3, MPO, GBM and dsDNA (CytoBead® RPGN)

HEp-2 cells and microparticles coated with  La, CENP-B, Sm, RNP-Sm, dsDNA, Scl-70, Ro60 and Ro52 (CytoBead® ANA)

HEp-2 cells and microparticles coated with La, Jo-1, Sm, RNP-Sm, dsDNA, Scl-70, Ro60 and Ro52 (CytoBead® ANA 2)

Monkey tissue sections (esophagus) and microparticles coated with tTG, DG and a-IgA (CytoBead® CeliAK)

Result

Detection of IgG autoantibodies / IgA autoantibodies (CytoBead® CeliAK) in human serum

Incubation time

60 Minutes

Sample material

50 - 80 µl diluted serum

The CytoBead® principle combines a stepwise diagnostic approach of screening and confirmatory testing in one reaction. Preparation time, materials and patient samples can be saved.

Grossmann, K., Röber, N., Hiemann, R., Rödiger, S., Schierack, P., Reinhold, D., Laass, M.-W., Conrad, K. & Roggenbuck, D. Simultaneous detection of celiac disease-specific IgA antibodies and total IgA. Auto Immun Highlights. 2016 Dec; 7(1): 2.

 

Scholz, J., Grossmann, K., Knütter, I., Hiemann, R., Sowa,  M., Röber, N., Rödiger, S., Schierack, P., Reinhold, D., Bogdanos, D.P., Meroni, P.L., Radice, A., Conrad, K. & Roggenbuck, D. Second generation analysis of antinuclear antibody (ANA) by combination of screening and confirmatory testing. Clin Chem Lab Med 2015; 53(12): 1991-2002.

 

Sowa, M., Grossmann, K., Knütter, I., Hiemann, R., Röber, N., Anderer, U., Csernok, E., Bogdanos, D.P., Borghi, M.O., Meroni, P.L., Schierack, P., Reinhold, D., Conrad, K. & Roggenbuck, D. Simultaneous automated screening and confirmatory testing for vasculitis-specific ANCA. PLoS One. 2014; 9(9): e107743.

 

Sowa, M., Grossmann, K., Scholz, J., Röber, N., Rödiger, S., Schierack, P., Conrad, K., Roggenbuck, D. & Hiemann, R. The CytoBead assay - a novel approach of multiparametric autoantibody analysis in the diagnostics of systemic autoimmune diseases. J Med Lab 2014; 38: 309-17.