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Related Experiment Videos

A sensitive solid phase microradioimmunoassay for anti-double stranded DNA antibodies

F Fish, M Ziff

    Arthritis and Rheumatism
    |March 1, 1981
    PubMed
    Summary

    A new radioimmunoassay accurately measures anti-double stranded DNA (dsDNA) antibodies. This method utilizes poly-L-lysine to bind dsDNA to plastic, showing high specificity for detecting antibodies in systemic lupus erythematosus (SLE) patients.

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    Area of Science:

    • Immunology
    • Biochemistry
    • Molecular Biology

    Background:

    • Anti-double stranded DNA (dsDNA) antibodies are key biomarkers for systemic lupus erythematosus (SLE).
    • Accurate and sensitive detection of these antibodies is crucial for SLE diagnosis and monitoring.
    • Existing assays may have limitations in sensitivity or specificity.

    Purpose of the Study:

    • To develop and validate a sensitive solid-phase microradioimmunoassay for quantifying anti-dsDNA antibodies.
    • To utilize poly-L-lysine (PLL) for efficient binding of dsDNA to plastic surfaces.
    • To assess the assay's performance in distinguishing between SLE patients and individuals with other autoimmune conditions or healthy controls.

    Main Methods:

    • A solid-phase microradioimmunoassay was developed using poly-L-lysine to coat microtitration trays with dsDNA.

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  • Synthetic DNA (poly dA-dT) served as a model for dsDNA, and its suitability was confirmed by nuclease S1 digestion.
  • Patient and control sera were incubated on coated trays, and anti-dsDNA immunoglobulin (Ig) was detected using 125I-labeled anti-Ig.

    • Main Results:

    • The assay demonstrated high sensitivity and specificity for detecting anti-dsDNA antibodies.
    • Background binding was minimized by optimizing serum dilution and using blocking agents (BGG and BSA).
    • Significant correlation was observed between the radioimmunoassay results and those from the Crithidia luciliae assay.

    Conclusions:

    • The developed radioimmunoassay is a sensitive and specific method for measuring anti-dsDNA antibodies.
    • The assay effectively identified anti-dsDNA activity in active SLE patients, with low detection rates in other conditions.
    • This assay shows promise as a valuable tool for SLE diagnosis and patient management.