Validation of a commercially available kit to detect anti-phosphatidylserine/prothrombin antibodies in a cohort of systemic lupus erythematosus patients

Thromb Res. 2014 Mar;133(3):451-4. doi: 10.1016/j.thromres.2013.12.001. Epub 2013 Dec 7.

Abstract

Background: Antiprothrombin antibodies detection comprises two different ELISAs: prothrombin coated on irradiated plates (aPT) or phosphatidylserine/prothrombin (aPS/PT) as the antigen. While several commercial kits are available for the detection of aPT, aPS/PT are usually detected by in-house assays. Recently, a new commercially available kit was launched and, therefore, we decided to test its efficiency by comparing it to our in-house assay.

Methods: aPS/PT were tested by our in-house assay (aPS/PT(ih)) in 75 SLE patients, using Immulon 1 plates coated with phosphatidylserine, purified human prothrombin and 1%BSA-TBS-CaCl as blocking and diluents. Data from this assay were compared to those obtained by the QUANTA Lite aPS/PT screen, IgG and IgM Elisa (INOVA Diagnostics, Inc, San Diego, USA) commercial kits (aPS/PT(c)).

Results: aPS/PT were found in 41.3% and 46.7% of SLE patients by the aPS/PT(ih) and the aPS/PT(c), respectively. There was a positive correlation between IgG aPS/PT(ih) and aPS/PT(c) assays (R(2)=0. 861 by Spearman test, p=0.0027). Sensitivity and specificity for APS were 62.2% and 97.4% (AUC 0.780) for the aPS/PT(ih) assay and 70.3% and 84.2% (AUC 0.858) for the aPS/PT(C). Shorter running times were also seen when comparing the aPS/PT(ih) vs. aPS/PT(c) (7hours vs. 3hours, respectively).

Conclusion: The aPS/PT(C) is a reproducible and accurate test for the detection of aPS/PT, bringing also the advantage of shorter running times.

Keywords: QUANTA Lite™ aPS/PT; antiphospholipid syndrome; phosphatidylserine-prothrombin complex; prothrombin.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Adult
  • Antibodies, Antiphospholipid / analysis*
  • Antibodies, Antiphospholipid / blood
  • Antibodies, Antiphospholipid / immunology
  • Antiphospholipid Syndrome / blood
  • Antiphospholipid Syndrome / immunology
  • Biomarkers / blood
  • Cohort Studies
  • Female
  • Humans
  • Lupus Erythematosus, Systemic / blood
  • Lupus Erythematosus, Systemic / immunology*
  • Male
  • Phosphatidylserines / immunology*
  • Predictive Value of Tests
  • Prothrombin / immunology*
  • Reagent Kits, Diagnostic / standards*

Substances

  • Antibodies, Antiphospholipid
  • Biomarkers
  • Phosphatidylserines
  • Reagent Kits, Diagnostic
  • Prothrombin