Background Antiphospholipid antibodies (aPL) are associated with arterial/venous thrombosis and are detected most often in patients with antiphospholipid syndrome (APS). APS is an autoimmune disorder comprising arterial or venous thrombosis, thrombocytopenia and recurrent foetal loss and is characterized by the appearance of autoantibodies to negatively charged phospholipids (1). The main goal of our study was to conduct an immunodot assay of IgG and IgM antibodies to phospholipids (cardiolipin, phosphatidyl-serine and phosphatidyl-inositol) and β2-glycoprotein I in patients with arterial and venous thrombosis as screening method for APS with the following verification of APS by Enzyme-Linked Immunosorbent Assay (ELISA).
Objectives We examined 21 patients with arterial thrombosis (6 males and 15 females, age 38,0 years (lower quartile 26,0, upper quartile 48,0)) and 19 patients with venous thrombosis (19 females, age 36,0 years (lower quartile 24,0, upper quartile 45,0)).
Methods We performed qualitative detection of IgG and IgM antibodies to phospholipids (cardiolipin (aCL), phosphatidyl-serine (aPhS) and phosphatidyl-inositol (aPhI)) and β2-glycoprotein I (aβ2-GP-1) using Anti-Phospholipid Dot (GA GENERIC ASSAYS GmbH) according to the instructions of manufacturer. Each Anti-Phospholipid Dot kit includes 20 numbered strips. The strips consist of a membrane where 4 different autoantigenes lines and a positive control line sprayes on. IgG and IgM aCL, aβ2-GP-1 were assessed by Immunometric Enzyme Immunoassay for the quantitative determination (Orgentec, Germany) according to the instructions of manufacturer.
Results The patients with arterial and venous thrombosis had positive IgG aCL (100% and 100%, respectively) and IgG aβ2-GP-1 (90,5% (95% CI: 77,9-100%) and 78,9% (95% CI: 60,6-97,3%) respectively). It has also been detected that both groups of patients had increased frequency of occurence of positive IgG aPhS (76,2% (95% CI: 57,9-94,4%), (73,7% (95% CI: 53,9-93,5%) respectively) and aPhI (71,4% (95% CI: 52,1-90,8%), (68,4% (95% CI: 7,5-89,3%) respectively). High occurence of positive IgM aCL, IgM aβ2-GP-1 and IgM aPhS were revealed in patients with arterial thrombosis. The difference between frequency of positive IgG aPhS and IgG aPhI in patients with venous thrombosis was not significant. The frequency of positive IgM aβ2-GP-1 in patients with arterial thrombosis (81% (95% CI: 64,2-97,8%)) was higher than in patients with venous thrombosis (47,4% (95% CI: 24,9-69,8%)) (χ2=0,026). Increased levels of IgG and IgM aCL, aβ2-GP-1 revealed by Anti-Phospholipid Dot were confirmed with the help of ELISA.
Conclusions All studied patients with arterial and venous thrombosis demonstrated increased levels of antiphospholipid antibodies in the whole and the highest frequency of positive IgG aCL and aβ2-GP-1. Qualitative detection of IgG and IgM antibodies to phospholipids with the help of Anti-Phospholipid Dot (GA GENERIC ASSAYS GmbH) can be used to screen the patients with arterial and venous thrombosis for APS.
Myakis S., Lockshin M.D., Atsumi T. et al. International consensus statement on an update of the classification criteria for definite antiphospholipid syndrome. J Thromb and Haemost 2006;4:295-306.
Disclosure of Interest None Declared
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