Method Description

SP100 and GP210

These tests are intended for the semi-quantitative detection of anti-gp210 or anti-Sp100 antibody of the IgG class in human serum. A purified peptide corresponding to a portion of the gp210 or Sp100 protein is bound to the wells of a polystyrene microwell plate. Pre-diluted controls and diluted patient sera are added to separate wells, allowing any gp210 or Sp100 antibodies present to bind to the immobilized antigen. Unbound sample is washed away, and an enzyme labeled anti-human IgG conjugate is added to each well. A second incubation allows the enzyme labeled anti-human IgG to bind to any patient antibodies, which have become attached to the microwells. After washing away any unbound enzyme labeled anti-human IgG, the remaining enzyme activity is measured by adding a chromogenic substrate and measuring the intensity of the color that develops. The assay can be evaluated spectrophotometrically by measuring and comparing the color intensity that develops in the patient wells with the control in the control wells.(Package inserts: QUANTA Lite gp210 ELISA 708995. INOVA Diagnostics; Rev. 5, 04/2019; QUANTA Lite sp100 ELISA 708990. INOVA Diagnostics; Rev. 3, 12/2018).

Antinuclear Antibodies, Hep-2 Substrate,

Antibodies to nuclear antigens in a human epithelial type 2 (HEp-2) cell line by an indirect immunofluorescent technique. Commercial slides prepared from HEp-2 cells are used as a substrate. IgG antibodies in serum specimens are detected after incubation of serum with the commercial slides by the addition of a fluorescein isothiocyanate (FITC)-labeled antihuman-IgG reagent. All patient specimens are initially screened at 1:80.(Package insert: NOVA Lite DAPI ANA. Inova Diagnostics; 05/2015)

Mitochondrial Antibodies, M2

A recombinant pyruvate dehydrogenase complex-E2 (M2) antigen for detection of antibodies against M2 is attached to the surface of a microplate. Diluted patient serum, standards, or controls are added to the wells, and the M2 specific IgG and IgM antibodies, if present, bind to the antigen. All unbound human antibodies are washed away, and a conjugate of enzyme-labeled polyclonal antibody to human IgG and IgM is added. The enzyme conjugate binds to the antibody complex. Excess enzyme-conjugate is washed away, and substrate is added. After a specified time, the enzyme reaction is stopped. The intensity of the color generated is proportional to the amount of anti-M2 IgG and/or IgM antibody in the sample. The results are read by a spectrophotometer producing a direct measurement of the anti-M2 IgG and IgM antibodies in the serum. Testing is performed on the Agility instrument by Dynex.(Package insert: Kallestad Anti-Mitochondrial Kit. Bio-Rad Laboratories, Inc; 04/2014)