Antinuclear Antibodies (ANA), Serum
Method Description Describes how the test is performed and provides a method-specific reference
The method used to detect antinuclear antibody (ANA) is enzyme-linked immunosorbent assay (ELISA). A HEp-2 lysate supplemented with various purified antigens (double-stranded deoxyribonucleic acid (dsDNA), histone, SS-A (Ro), SS-B (La) Smith, RNP, Scl-70, Jo-1, plus centromere antigen) are coated onto microtiter plate wells. A dilution of patient serum is added to the well and incubated. After washing to remove unbound serum protein, an enzyme conjugated antihuman IgG antibody is added to detect human IgG bound to the microtiter plate well. After incubation and washing to remove unbound conjugate, a substrate to the enzyme is added to the well. After incubation, the enzyme substrate reaction is stopped. The complete assay is measured on a spectrophotometer plate reader. The optical density measured is proportional to the antibody present in the patient serum. Testing is performed on the Triturus instrument by Grifols.(Package insert: ELISA kits, Bio-Rad Laboratories, Hercules CA)
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