Please use this identifier to cite or link to this item: http://ahro.austin.org.au/austinjspui/handle/1/9156
Title: Detection of anti-phosphatidylethanolamine antibodies using flow cytometry.
Authors: Drouvalakis, K A;Neeson, P J;Buchanan, Russell R C
Affiliation: Department of Medicine, The University of Melbourne, Austin & Repatriation Medical Centre, Heidelberg, Victoria, Australia
drouvaka@leland.stanford.edu
Issue Date: 1-May-1999
Citation: Cytometry; 36(1): 46-51
Abstract: The finding that lupus anticoagulant (LA) is significantly associated with anti-phosphatidylethanolamine (PE) activity has led to great interest in its relation to the clinical features of the antiphospholipid syndrome (APS). Considerable variability has, however, been reported in the prevalence of anti-PE antibodies in APS patients using enzyme-linked immunosorbent assay (ELISA) methodology. The lack of standardization and differences in technique may in part explain these discrepancies. PE binds variably to different types of microtiter wells, reflected in the consequent detection, or lack of detection, of anti-PE antibodies. This study describes the use of flow cytometry as an alternative method for the detection of anti-PE antibodies.Six LA-positive plasma samples were used in this original study. Polystyrene beads were coated with PE overnight. These were subsequently incubated with patient plasma. Both IgG and IgM binding were detected by flow cytometry using a cocktail of fluorescently labelled anti-human Ig isotypes.When these results were compared with those from ELISA, flow cytometric analysis provided an apparent enhanced detection of anti-PE antibodies. It was found that 6/6 were IgM anti-PE positive by flow cytometry, whereas 5/6 were IgM by ELISA; 2/6 negative for anti-cardiolipin antibodies by ELISA were positive by flow cytometry; and 2/6 positive for antiphosphatidylcholine antibodies in cytometry were negative by ELISA.With appropriate quantification, this method may be more sensitive than ELISA in detecting anti-PE antibodies in plasma samples of patients with APS.
Internal ID Number: 10331626
URI: http://ahro.austin.org.au/austinjspui/handle/1/9156
URL: http://www.ncbi.nlm.nih.gov/pubmed/10331626
Type: Journal Article
Subjects: Antibodies, Antiphospholipid.analysis.blood
Autoimmune Diseases.diagnosis.immunology
Enzyme-Linked Immunosorbent Assay.methods
Flow Cytometry.methods
Humans
Microspheres
Phosphatidylethanolamines.immunology
Polystyrenes
Appears in Collections:Journal articles

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