The Development of a Semi-automated Latex Agglutination Test for the Detection of Antibodies to Anaplasma marginale Using a Cell Culture-derived Antigen

Serologic diagnosis of anaplasmosis is currently done by the complement‐fixation, ELISA, and card agglutination tests. These tests have utilized A. marginale harvested from bovine erythrocytes as antigen which is often contaminated with erythrocyte stroma. We are currently testing A. marginale propa...

Full description

Saved in:
Bibliographic Details
Published in:Annals of the New York Academy of Sciences 1998-06, Vol.849 (1), p.282-292
Main Authors: RODGERS, S. J., SALIKI, J. T., BLOUIN, E. F., KOCAN, K. M.
Format: Article
Language:English
Subjects:
Anaplasma - immunology
Anaplasma - isolation & purification
Anaplasma marginale
Anaplasmosis - diagnosis
Animals
Antibodies, Bacterial - blood
Antigen-Antibody Reactions
Automation
Cattle
Cells, Cultured
Enzyme-Linked Immunosorbent Assay
Erythrocytes - microbiology
Ixodes scapularis
Ixodidae
Latex Fixation Tests - instrumentation
Latex Fixation Tests - methods
Reproducibility of Results
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Serologic diagnosis of anaplasmosis is currently done by the complement‐fixation, ELISA, and card agglutination tests. These tests have utilized A. marginale harvested from bovine erythrocytes as antigen which is often contaminated with erythrocyte stroma. We are currently testing A. marginale propagated in a Ixodes scapularis cell line as antigen for serologic tests. In this study, we report the use of the cell culture‐derived A. marginale as antigen for development of a rapid, semi‐automated latex agglutination test. Diluted serum and latex (polystyrene microspheres), sensitized with cell culture‐derived A. marginale proteins, were dispensed into 96‐well microtiter plates. An initial reading of light transmission was recorded by a computer‐interfaced scanning autoreader. After 30 minutes, the plates were mixed and read a second time, recording the delta % light transmittance. The sensitized latex microspheres (latex) agglutinated in the presence of A. marginale antibodies, thus producing an increase in light transmittance. In preliminary tests, 724/977 of the sera were positive for A. marginale antibodies with an apparent agreement of 83.3% when compared with the complement‐fixation test. Sensitization and sera dilution buffers were shown to have a marked effect on the sensitivity and specificity of this assay. Results will be presented on the optimization of buffers and the testing of sera from experimentally and field‐infected cattle.
ISSN:0077-8923
1749-6632
DOI:10.1111/j.1749-6632.1998.tb11060.x