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CIEDia - a new technology for the detection of precipitating antibodies. Quicker, more sensitive, easier to operate and evaluate than traditional gel-based assays. When applied to autoimmune diagnostics:

  • effective in detecting precipitating autoantibodies to: RNP/Sm, Sm, SSA, SSB, Jo-1 and Scl-70
  • improved diagnostic specificity over alternative assays
  • improved positive predictive value (PPV) for autoimmune disease
  • resolves samples yielding "equivocal" results with alternative assays
  • confirmation/rejection of positive results arising from entry diagnostic procedures

CIEDia (patent pending) - a collaboration between Gemini Research Limited (New Zealand) and AROTEC Diagnostics

PRINCIPLE OF THE METHOD

CIEDia is a technology based on counterimmunoelectrophoresis (CIE) whereby an electric field is used to induce migration of an antigen and antibodies towards one another in such a way that a precipitin line is formed should a recognition event occur. Traditional CIE assays have the disadvantages of requiring post-electrophoretic staining /destaining,  difficulty in evaluating when formation of precipitin is complete and require co-electrophoresis of antibody standards to determine specificity. CIEDia circumvents all these issues by utilising highly pure fluorescently labelled antigen. This permits the real-time detection of specific precipitin formation with greatly improved sensitivity. Precipitin formation requires a polyclonal immune response and can therefore be considered more stringent than other assays with regard to yielding a positive result.

HOW IT OPERATES

Equipment required:

  • blue light transilluminator (Invitrogen) f

  • horizontal electrophoresis power supply (Invitrogen) e

  • adapter (CIEDia*) d

  • agarose gel (CIEDia*) c

  • buffer strips (CIEDia*) b

  • amber filter (Invitrogen) a

Procedure:

  • assemble equipment components b-f

  • apply patient samples and fluorescently labelled antigen

  • mount amber filter and commence electrophoresis

  • observe precipitin formation through the amber filter after activating the transilluminator

The antigen is labelled with a fluorophore that is excited by the blue light transilluminator. The light emitted by the fluorophore (but not the blue light) will penetrate the amber filter where it can be visually observed.

APPLICATIONS

Rapid and sensitive detection of precipitating SSA autoantibodies.
Top panel: time course of the formation of fluorescent immunoprecipitate during immunoelectrophoresis. Middle panel: detection of precipitating SSA autoantibodies at different serum dilutions. Bottom panel: specificity of the CIEDia assay; only patient samples containing SSA antibodies are able to form an immunoprecipitate.
Literature: Hood, KA et al. Poster C-134, AACC Annual Meeting, Los Angeles 2012

 

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