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Phospholipid Screen IgG/IgM ELISA Kit

Full Name: Phospholipid Screen IgG/IgM ELISA Kit
Reactivity: Human
Sample Type: Plasma, Serum
Sensitivity: 0.5 U/ml

BACKGROUND

Phospholipid antibodiеs arе found in Systеmic Lupus Erythеmatosus (SLE), thеy havе also bееn found in somе cancеrs and HIV. Cardiolipin antibodiеs and lupus anticoagulant arе thе most common anti-phospholipid antibodiеs, othеrs that havе also bе analysеd includе; anti-phosphatidylsеrinе and anti-bеta2 glycoprotеin.

Phospholipid antibodiеs can bе еithеr tеstеd for bеta-2 glycoprotеin I or cardiolipin (this involvеs thе dеtеction of sеvеral classеs IgA, IgG, and/or IgM) or lupus anticoagulant assays (a functional tеst that mеasurеs timе takеn for patiеnt samplе to clot, phosopholipids play an important part in thе clotting procеss).

INTENDED USE

Human phospholipid screen IgG/IgM ELISA kit is intended for screening in-vitro quantitative levels of IgG and IgM auto-antibodies to beta-2 glycoprotein I and anti phospholipid (phospholipid IgG/IgM) in human serum and plasma samples. This assay has a minimum analytical sensitivity limit of 0.5 U/ml.

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:Highly purified mixture of human beta-2 glycoprotein I, cardiolipin, phosphatidyl inositol, phosphatidic acid and phosphatidyl serine is bound to microwells.
  • Calibrator A-F: Concentration 0, 6.3, 12.5, 25, 50, 100 U/ml, made up from phospholipid antibody.
  • Control Positive and Control Negative: Containing phospholipid antibodies in a serum/buffer matrix.
  • Sample Buffer P (5x).
  • Enzyme Conjugate: Contains anti-human IgG antibodies, HRP labelled.
  • Enzyme Conjugate: Contains anti-human IgM antibodies, HRP labelled.
  • TMB Substrate.
  • Stop Solution: Contains acid.
  • Wash Buffer (50x).
  • Instruction for Use.
  • Certificate of Analysis.

SENSITIVITY

The minimum detection sensitivity level of IgG and IgM autoantibodies to screen phospholipids IgG/IgM and beta-2-glycoprotein I using current phospholipid IgG or IgM ELISA kit was 0.5 U/ml. The dynamic range for this assay is 6.3 – 100.0 U/ml.

ASSAY CHARACTERISTICS

– Measuring Range: IgG (0 – 100 GPL-U/ml), IgM (0 – 100 MPL-U/ml).
– Expected Values: Samples from healthy blood donors displayed a cut-off IgG at 10 GPL-U/ml, cut-off IgM at 10 MPL-U/ml.
– Results Interpretation: IgG (Negative < 10 GPL-U/ml, Positive ≥ 10 GPL-U/ml), IgM (Negative < 10 MPL-U/ml, Positive ≥ 10 MPL-U/ml).
– Linearity: IgG (94 – 101%), IgM (95 – 101%).
– Limit Of Detection (LOD): Functional sensitivity was determined to be IgG (0.5 GPL-U/ml), IgM (0.5 MPL-U/ml).
– Intra Assay: IgG 3.4 – 5.2%, IgM 3.5 – 5.1%
– Inter Assay: IgG 3.6 – 5.4%, IgM 4.1 – 5.3%
– Clinical Diagnosis: IgG: Sensitivity (91.8%), Specificity (97.3%), Overall agreement (95.5%), IgM: Sensitivity (53.4%), Specificity (96.7%), Overall agreement (82.5%).

REFERENCES

  1. Coagulation assays for the lupus anticoagulant: review and critique of current methodology. Stroke. (1992) 23 (2 Suppl): I11-4. Review. Triplett D.A.
  2. The anti-phospholipid antibody syndrome (Hughes syndrome): therapeutic aspects. Baillieres Best Pract Res Clin Rheumatol. (2000) 14 (1): 151-63. Cuadrado M.J. and Khamashta M.A.
  3. Surprising roles for phospholipid binding proteins revealed by high throughput genetics. Biochem Cell Biol. (2010) 88 (4): 565-74. Review. LeBlanc M.A. and McMaster C.R.
  4. Autoantibodies to phospholipids–new looks at old diseases–a primer for physicians. J Okla State Med Assoc. (1992) 85 (2): 61-8. Review. McCarty G.A.

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