Rapid detection of anticardiolipin antibodies

Abstract

A rapid screening method for the detection of antiphospholipid antibodies is described. Dense, red dyed polystyrene beads coated with cardiolipin were incubated with test sera for a short period of time, then added to a microtube containing anti-human IgG in a gel provided within a precast card (DiaMed ID Microtyping System). The card was centrifuged at 150g for 5 min and then examined for movement of the beads through the gel. Beads without bound antibody travelled through the gel and formed a pellet on the bottom of the tube. Anti-human IgG within the gel matrix impeded cardiolipin-coated beads when antiphospholipid antibodies bound to the beads. Positivity was indicated by the formation of a layer of beads on the top of the gel matrix. Prospective analysis of 103 samples for the presence of antiphospholipid antibodies by flow cytometry and the gel-card technique showed good correlation between the two methods. All samples found to be positive by flow cytometry (23 of 103) were identified as positive by the gel-card technique. Two samples were identified as positive by the gel-card method but negative by flow cytometry. The technique is simple to perform and should prove useful as a rapid screening method for the detection of antiphospholipid antibodies. Am.

Cutaneous manifestations associated with antiphospholipid antibodies

“…Almost all patients with LA have detectable aCL antibodies (90%) and almost all patients with AAS have detectable aCL antibodies 3 . Low protein C and protein S levels are described in patients with AAS and are associated with an additional risk of thrombotic events 28–32 …”

“…If patients referred any symptoms of possible associated conditions, further laboratory tests were performed in an attempt to diagnose the condition. Patients with more systemic complications 7 and greater risk of recurrence were also tested for deficiency of protein S, protein C and resistance to activated protein C, and had their antithrombin III and fibrinogen levels measured 28–32 . Histologic samples from skin lesions (nodules, cutaneous necrosis and ulcers) was collected, fixed in formalin, processed in paraffin and stained with hematoxylin‐eosin 46 .…”

“…Lupus anticoagulant is measured by diluted Russell's viper venom time (dRVVT) or Kaolin clotting time (KCT). Low levels of reactive C protein, protein C and protein S indicate additional risk of thrombosis 28–32 …”

Antiphospholipid Antibodies: Laboratory and Pathogenetic Aspects

“…Other methods that have been used to detect aPL include ELISAs for the detection of antibodies against other PLs (e.g., PS, phosphatidylethanolamine) 159 or against a mixture of PLs (aPL ELISAs), 160,161 cardiolipin-coated microbeads, 162 flow cytometry, 163,164 and a chromogenic thrombin production assay that has been reported to be more sensitive than LA. 165 …”

Rapid detection of autoantibodies to dsDNA with the particle gel immunoassay (ID-PaGIA)

“…[5][6][7][8][9] ID card The ID-PaGIA assay format is adapted for the standard methodology and equipment of the ID-Micro Typing system using ID cards with buffer compositions specially adjusted to the requirements of the synthetic particles. The buffer used in the ID-dsDNA antibody test also contained rabbit antihuman antiglobulin serum.…”