Phospholipid Screen IgG/IgM ELISA Kit
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.
BACKGROUND
Phospholipid antibodies are often found in autoimmune disorders (such as Systemic Lupus Erythematosus, SLE), they have also been found in some cancers and HIV. Cardiolipin antibodies and lupus anticoagulant are the most common anti-phospholipid antibodies, others that have also be analysed include anti-phosphatidylserine and anti-beta2 glycoprotein. Phospholipid antibodies can be either tested for beta-2 glycoprotein I or cardiolipin (this involves the detection of several classes IgA, IgG, and/or IgM) or lupus anticoagulant assays (a functional test that measures time taken for patient sample to clot, phosopholipids play an important part in the clotting process). There are a number of studies which have indicated a correlation between these autoantibodies and an enhanced incidence of thrombosis and thrombocytopenia.
PHOSPHOLIPID SCREEN IGG/IGM 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: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/IgM ELISA kit was 0.5 U/ml. The dynamic range for this assay is 6.3 – 100.0 U/ml.
ASSAY CHARACTERISTICS
– Phospholipid Screen (Phospholipid IgG/IgM): ELISA
– 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
- Coagulation assays for the lupus anticoagulant: review and critique of current methodology. Stroke. (1992) 23 (2 Suppl): I11-4. Review. Triplett D.A.
- 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.
- 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.
- 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.
ADDITIONAL INFORMATION
- Full Name: Phospholipid Screen IgG/IgM ELISA Kit
- Reactivity: Human
- Sample Type: Plasma, Serum
- Sensitivity: 0.5 U/ml
