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Anti-ssDNA Ab ELISA Kit

INTENDED USE

Human anti-ssDNA antibody ELISA kit is intended for analysing in-vitro quantitative concentrations of anti IgG auto-antibodies to single stranded DNA (ssDNA Ab, anti-ssDNA antibody) in human plasma and serum samples. This assay has a minimum analytical sensitivity limit of 1.0 U/ml.

BACKGROUND

Auto-antibodies against DNA are usually either IgM or IgG type which are able to react with natural single-stranded DNA (ssDNA), double stranded DNA (dsDNA) or both types. Generally, ssDNA antibodies tend to be found in high frequency in patients that either have systemic sclerosis or systemic lupus erythematosus, they are also present in both non-rheumatic and rheumatic diseases. Single stranded DNA antibodies are vital in conjunction with detecting histone antibodies for the differential diagnosis of drug-induced lupus. In the serum of SLE patients, up to 87% patients display ssDNA antibodies during the acute relapse of the disease whereas only 43% is present in the inactive phase. There is also evidence to indicate that ssDNA antibodies are present in patients with juvenile chronic arthritis (JCA) and collagenases.

ANTI-SSDNA AB ELISA KIT CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

  • Divisible Microplate: Consisting of 12 modules of 8 wells, recombinant single stranded DNA (ssDNA) is bound to microwells.
  • Calibrator A-F: Concentration 0, 0, 12.5, 25, 50, 100, 200 U/ml, made up from anti-ssDNA Ab.
  • Control Positive And Control Negative: Containing human ssDNA antibody in a serum/buffer matrix.
  • Sample Buffer P (5x).
  • Enzyme Conjugate: Contains anti-human IgG antibodies, HRP labelled.
  • Substrate: Tetramethylbenzidine (TMB).
  • Stop Solution: Contains acid.
  • Wash Buffer (50x).

SENSITIVITY

The minimum detection sensitivity level of IgG antibodies to human single stranded DNA (ssDNA Ab, anti-ssDNA antibody, anti-ssDNA Ab) using current human anti-ssDNA antibody ELISA kit was 1.0 U/ml. The dynamic range for this assay is 12.5 – 200.0 U/ml.

ASSAY CHARACTERISTICS

– Anti-Single Stranded DNA Antibody (Anti-ssDNA Ab): ELISA
– Measuring Range: 0 – 200 U/ml.
– Expected Values: Samples from healthy blood donors display a cut-off 20 U/ml.
– Results Interpretation: Negative: < 20 U/ml, Positive: ≥ 20 U/ml.
– Linearity: 95 – 106%
– Limit Of Detection (LOD): Functional sensitivity 1 U/ml.
– Intra Assay: 4.2 – 6.4%
– Inter Assay: 4.4 – 6.8%
– Interfering Substances: None
– Clinical Diagnosis: Sensitivity (35.4%), Specificity (98.4%), Overall agreement (85.8%).

REFERENCES

  1. Single-Stranded DNA Aptamers against Pathogens and Toxins: Identification and Biosensing Applications. Biomed Res Int. (2015) 2015: 419318. Review. Hong K.L. and Sooter L.J.
  2. Expression of antibody activity measured by ELISA. Anti-ssDNA antibody activity characterized by the shape of the dose-response curve. J Immunol Methods. (1983) 62 (3): 315-23. Gripenberg M. and Gripenberg G.
  3. Mutational analysis of a sequence-specific ssDNA binding lupus autoantibody. Biochemistry. (2003) 42 (1): 30-41. Cleary J. and Glick G.D.
  4. Immunoregulatory characteristics of the in vitro anti-ssDNA response. Immunol Res. (1993) 12 (4): 349-57. Anderson C.C., et al.
  5. Binding hot-spots in an antibody-ssDNA interface: a molecular dynamics study. Mol Biosyst. (2012) 8 (12): 3274-80. Wang Y.T. and Lee W.J.
  6. Thermodynamic basis for sequence-specific recognition of ssDNA by an autoantibody. Biochemistry. (2001) 40 (9): 2911-22. Ackroyd P.C., et al.

ADDITIONAL INFORMATION

  • Full Name: Anti-ssDNA Ab ELISA Kit
  • Reactivity: Human
  • Sample Type: Serum, Plasma
  • Sensitivity: 1.0 U/ml

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