FLUOROMETRIC IMMUNOASSAY FOR DETECTION OF ANTI-DSDNA ANTIBODIES

摘要

The present invention relates to the field of immunological methods, more precisely to the field of detection methods for antibodies against double-stranded DNA (dsDNA) for diagnostics of chronic autoimmune diseases, such as systemic lupus erythematosus (SLE). The fluorometric immunoassay method for detection of anti-dsDNA solves the technical problem of designing a method for detection of the aforesaid antibodies, which would be faster, cheaper and less toxic as the standard Farr-RIA method, but would have the same diagnostic specificity (which is 100%) and improved diagnostic sensitivity (for 3%). Detection of anti-dsDNA is based on detection of fluorescence in two fractions of samples, in the supernatant and in the sample with a precipitate, which contains immune complexes composed of added dsDNA and anti-dsDNA present in the patient's serum, wherein the detected fluorescence is a consequence of binding fluorescent dyes with dsDNA bound in the complexes or with free dsDNA. The method according to the invention allows obtaining reliable results, fast sample analysis and results availability to the attending physician, use of human and environmentally safe chemicals and lowering of costs in comparison to the standard Farr-RIA method.

主权项

1. A fluorometric immunoassay method for detecting antibodies against double-stranded DNA (anti-dsDNA), comprising
precipitation of immune complexes with a saturated solution of ammonium sulfate or solution of polyethylene glycol (PEG), wherein the immune complexes are a consequence of incubation of decomplemented patient's serum with dsDNA in a warm place, detection of fluorescence in two fractions of samples, namely in supernatant and in the sample with precipitate, comprising immune complexes comprising dsDNA and anti-dsDNA, wherein the detected fluorescence is a consequence of binding of fluorescent dyes into the dsDNA.