Abstract

BackgroundAntiphospholipid antibodies (aPL) can be detected in asymptomatic carriers and infectious patients. The aim was to investigate whether a novel line immunoassay (LIA) differentiates between antiphospholipid syndrome (APS) and asymptomatic aPL+ carriers or patients with infectious diseases (infectious diseases controls (IDC)).MethodsSixty-one patients with APS (56 primary, 22/56 with obstetric events only, and 5 secondary), 146 controls including 24 aPL+ asymptomatic carriers and 73 IDC were tested on a novel hydrophobic solid phase coated with cardiolipin (CL), phosphatic acid, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylserine, beta2-glycoprotein I (β2GPI), prothrombin, and annexin V. Samples were also tested by anti-CL and anti-β2GPI ELISAs and for lupus anticoagulant activity. Human monoclonal antibodies (humoAbs) against human β2GPI or PL alone were tested on the same LIA substrates in the absence or presence of human serum, purified human β2GPI or after CL-micelle absorption.ResultsComparison of LIA with the aPL-classification assays revealed good agreement for IgG/IgM aß2GPI and aCL. Anti-CL and anti-ß2GPI IgG/IgM reactivity assessed by LIA was significantly higher in patients with APS versus healthy controls and IDCs, as detected by ELISA. IgG binding to CL and ß2GPI in the LIA was significantly lower in aPL+ carriers and Venereal Disease Research Laboratory test (VDRL) + samples than in patients with APS. HumoAb against domain 1 recognized β2GPI bound to the LIA-matrix and in anionic phospholipid (PL) complexes. Absorption with CL micelles abolished the reactivity of a PL-specific humoAb but did not affect the binding of anti-β2GPI humoAbs.ConclusionsThe LIA and ELISA have good agreement in detecting aPL in APS, but the LIA differentiates patients with APS from infectious patients and asymptomatic carriers, likely through the exposure of domain 1.Electronic supplementary materialThe online version of this article (doi:10.1186/s13075-016-1018-x) contains supplementary material, which is available to authorized users.

Highlights

  • Antiphospholipid antibodies can be detected in asymptomatic carriers and infectious patients

  • We speculated on whether line immunoassay (LIA) demonstrating improved performance characteristics in comparison with enzyme-linked immunosorbent assay (ELISA) can be the first test able to detect aPL assisting in the differentiation of patients with antiphospholipid syndrome (APS) from asymptomatic aPL+ carriers and aPL-positive patients with infectious diseases

  • As disease controls we included 73 patients suffering from infectious diseases (infectious diseases controls (IDC)): 3 of these patients were infected with Epstein-Barr virus, 14 with Toxoplasma gondii, 24 with cytomegalovirus (CMV), 8 with Rubella virus, 1 with hepatitis C and 23 with Treponema pallidum displaying a positive Venereal Disease Research Laboratory test result (VDRL+))

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Summary

Introduction

Antiphospholipid antibodies (aPL) can be detected in asymptomatic carriers and infectious patients. The aim was to investigate whether a novel line immunoassay (LIA) differentiates between antiphospholipid syndrome (APS) and asymptomatic aPL+ carriers or patients with infectious diseases (infectious diseases controls (IDC)). The current international consensus for the classification of APS values clinical and laboratory criteria for the diagnosis of APS [2]. The latter criterion comprises the detection of persistent aPL by solid-phase assays, i.e., IgG and IgM to beta2 - glycoprotein I (β2GPI) and the cardiolipin (CL)-β2GPI complex, and by a functional clotting test, i.e., the lupus anticoagulant (LA). The relevance of aPL assay techniques involving the interaction of PL-binding proteins like β2GPI and PT with PLs other than CL, such as phosphatidylserine (PS), is still a matter of debate and not yet included in the classification criteria [13]

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