Microarray ELISA for Autoantibody Screening in Connective Tissue Diseases 200-206
Dr. K.D. Kumble, Pictor Limited, 24 Balfour Road,
Parnell, Auckland 1052, New Zealand.
Phone: +64 9 309 0950
This study was performed to demonstrate the use of an ELISA-based microarray technology which is termed as ‘PictArrays’, to identify autoantibody expression patterns in patients with symptoms of autoimmune connective tissue disease.
Eight commonly tested antigens were simultaneously tested on specially designed 16-well slides for their autoantibody expression patterns. The assay specificity, sensitivity and reproducibility for each of the antigens were measured. The results were analyzed by using specially developed algorithms to identify seropositive samples.
The multiplex assay could identify specific antigen binding by autoimmune sera on the arrays. The PictArray sensitivity was similar to that which was obtained in established immunoassays, and the assay reproducibility was within limits which were acceptable for diagnostic uses. The software could correctly identify the positive antigen reactivity at concentrations as low as 2 units/ ml of the antibody.
The data demonstrated the use of a multiplex platform to simultaneously measure multiple autoimmune antibodies. PictArrays offer significant advantages over other multiplex technologies, which include (i) the use of document scanners to read the test results (ii) ease of operation which requires no specialized technical training beyond that which is required for using the conventional ELISA kits (iii) reduction in errors through software-based data analysis, and (iv) inclusion of internal controls to monitor the assay performance of each sample. These features permit the use of PictArrays in resourceconstrained laboratories using existing infrastructure without significant capital expenditure.