A Solid Phase Enzyme‐linked Immunosorbent Assay (ELISA) for the Demonstration of Antibodies against Denatured, Single‐stranded DNA in Patient Sera

Gripenberg, Marianne; Linder, Ewert; Kurki, Pekka; Engvall, Eva

doi:10.1111/j.1365-3083.1978.tb00438.x

Cited by 53 publications

(13 citation statements)

References 19 publications

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“…Several investigators have attached DNA to untreated plastics (22)(23)(24)(25)(26)(27), although some reports have claimed that nDNA is not adsorbed to untreated polystyrene or polyvinyl (34,35). Pretreatment of plastics with poly-L lysine for radioimmunoassays (3 1.32) or protamine sulfate for EIA (25) has been successfully tried.…”

Section: Discussronmentioning

confidence: 99%

Enzyme immunoassay for antibodies to native DNA

Eaton

Schnneider

Schur

1983

Arthritis & Rheumatism

109

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An enzyme immunoassay was developed to detect antibodies to native DNA; DNA coating conditions that maximized sensitivity, specificity, and reproducibility were selected. Sera of patients with systemic lupus erythematosus (SLE) were positive more frequently by this immunoassay than by the Crithidiu luciliae assay or by counterimmunoelectrophoresis. By enzyme immunoassay, 94% of sera with active SLE and 70% of sera from patients with inactive SLE were positive, as were 16% from those suspected of having SLE, and 2.5% of normal persons. Specificity for native DNA was shown for both SLE and normal sera by inhibition studies and by S1 nuclease treatment of polystyrene-bound native DNA. The enzyme immunoassay correlated more with serum hemolytic complement levels than did the other 2 assays, suggesting that it detects biologically more relevant anti-DNA antibodies than do the other 2 tests.Measurement of serum anti-DNA antibody levels has been helpful in both the diagnosis and clinical assessment of systemic lupus erythematosus (SLE) detect antibodies to native DNA (nDNA), including immunodiffusion (7), complement fixation (8,9), agglutination (lO,ll), immunofluorescence (6,12), and radioimmunoassays (13-18). The sensitivity, specificity, cost, and ease of performance of these assays vary considerably (19,20).(The enzyme immunoassay (EIA) appears to offer advantages over other assays in areas of sensitivity, reproducibility, and efficiency, without sacrificing specificity. Since the initial description by Engvall and Perlmann (21), many antigens have been bound to solid phases to detect antibodies by EIA. Most of these antigens have been proteins or polysaccharides, but the binding of nucleic acids to polystyrene has not been systematically studied. Recently, several papers have appeared describing the application of EIAs to detect antibodies to DNA (22-29). In each of these reports different conditions were used, and the success of binding nDNA to the solid phase has varied (23,25).The purpose of the present study was to develop an EIA that could detect antibodies to nDNA, including a reproducible and reliable method for binding of DNA to the solid phase; this EIA was to be both sensitive and specific. In addition, the antibodies detected by the EIA were compared with those detected by other tests to define the (biologic) quality and relevance of the antibodies analyzed by different techniques. MATERIALS AND METHODSSource of sera. Normal human sera were obtained either from hospital personnel (sera kindly provided by Mr.

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Section: Discussronmentioning

confidence: 99%

Enzyme immunoassay for antibodies to native DNA

Eaton

Schnneider

Schur

1983

Arthritis & Rheumatism

109

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“…PM-1, Jo-1 and PCNA were detected by counterimmunoelectrophoresis [18]. The enzyme-linked immunosor bent assay (ELISA) was used for measurement of antissDNA antibody at a serum dilution of 1:200 [19], IgG and IgM ACA at 1:50 [20], and IgM RF at 1:100 [21]. Values were expressed as ELISA units (EU), obtained from the standard curve of each plate using running serial dilution of a positive control serum.…”

Section: Serological Methodsmentioning

confidence: 99%

Autoantibodies of Systemic Rheumatic Diseases in the Healthy Elderly

et al. 1990

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Antinuclear antibodies, anticardiolipin antibodies and IgM rheumatoid factor were determined in 300 healthy aged subjects, comparing their prevalence to that in 100 healthy subjects aged between 19 and 44 years and 352 patients under 65 years of age, affected by various systemic rheumatic diseases. Increased production of IgM rheumatoid factor and antinuclear and anticardiolipin antibodies was found as characteristic of aged humans, and suggested that a correction for age should be considered in evaluating the clinical significance of autoantibody profiles in elderly patients.

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“…In recent years, solid phase immunoassays have been developed which measure the interaction of antibody with insoluble D N A bound to a support medium (21)(22)(23)(24)(25)(26)(27) or incorporated within the kinetoplast of Crithidia luciliae (28,29), using labeled antibody to human gamma globulin. These tests appear to offer advantages over the conventional assays for anti-dsDNA antibodies.…”

Section: Clinical Significance Ofmentioning

confidence: 99%

Clinical Significance of Anti–double‐stranded Dna Antibodies Detected by a Solid Phase Enzyme Immunoassay

Miller

Lahita

Zarro

et al. 1981

Arthritis & Rheumatism

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A solid phase enzyme immunoassay (EIA) detected anti-double-stranded (ds) DNA antibodies in 88% of sera from patients classified clinically as having active systemic lupus erythematosus (SLE) without renal symptoms and 93% with renal disease. Fifty-six percent of sera from patients with inactive SLE were EIA positive for anti-dsDNA antibodies. The EIA had a sensitivity and specificity comparable to radioimmunoassay (RIA) and hemagglutination for patients with active SLE with or without renal disease, but it detected antidsDNA antibodies more frequently in patients with inactive SLE than the latter procedures. Precipitating antibodies detected by counterimmunoelectrophoresis (CIE) were less common in patients with renal disease (23% incidence) than clinically active patients without renal disease (7w0 incidence). Twenty-four SLE sera with elevated levels of Clq binding showed a 96% concordance for a positive EIA for antidsDNA antibodies in contrast to 66% concordance by RIA or hemagglutination. These findings suggest that the EIA is a sensitive and specific method for detection and measurement of anti-dsDNA antibodies. Several clinical applications of the EIA are discussed.

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A Solid Phase Enzyme‐linked Immunosorbent Assay (ELISA) for the Demonstration of Antibodies against Denatured, Single‐stranded DNA in Patient Sera

Cited by 53 publications

References 19 publications

Enzyme immunoassay for antibodies to native DNA

Enzyme immunoassay for antibodies to native DNA

Autoantibodies of Systemic Rheumatic Diseases in the Healthy Elderly

Clinical Significance of Anti–double‐stranded Dna Antibodies Detected by a Solid Phase Enzyme Immunoassay

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