Validation of multiplex microbead immunoassay for simultaneous serodetection of multiple infectious agents in laboratory mouse.

Authors

R Ravindran
I H. Khan
V V. Krishnan
M Ziman
L V. Kendall
J M. Frasier
R Bates
S M. Griffey
J R. Fahey
P A. Luciw

Document Type

Article

Publication Date

2010

Keywords

Antibodies-Bacterial, Antibodies-Viral, Antigens-Bacterial, Antigens-Viral, Bacterial-Infections, Immunoassay, Mice, Microspheres, Sensitivity-and-Specificity, Serologic-Tests, Virus-Diseases

First Page

51

Last Page

59

JAX Source

J Immunol Methods 2010 Dec; 363(1):51-9.

Abstract

Multiplex methodologies enable simultaneous detection of antibodies against several infectious agents allowing sample conservation, cost effectiveness, and amenability to high-throughput/automation. We have previously described a multiplex microbead immunoassay for serodetection of ten, high-priority mouse infectious pathogens. Here, we present a validation of this multiplex diagnostic system using approximately four hundred serum samples from different groups of mice. Computer assisted multivariate analysis of the resulting high volume data (8000 data points) was performed. This computational approach enabled presentation of data in a variety of easily interpretable formats (e.g., correlation tables and heat maps). Importantly, this computer aided approach was instrumental for the evaluation of assay accuracy, sensitivity, specificity, and robustness during the study. Crucial pieces of information were obtained to make timely adjustments for assay refinement. This progressive approach to developing an implementation-ready clinical assay, facilitated by computational analysis, produced a highly efficient, accurate and dependable serodiagnostics system. This system has effectively replaced the current state-of-the-art methodology (ELISA) used in mouse colony health management at the University of California and the Jackson Laboratory. A pathway to develop multiplex serology tests for infectious disease diagnosis described here serves as a model for multiplex immunoassay design, clinical validation, refinement and implementation.

Recommended Citation

Ravindran R, Khan IH, Krishnan VV, Ziman M, Kendall LV, Frasier JM, Bates R, Griffey SM, Fahey JR, Luciw PA. Validation of multiplex microbead immunoassay for simultaneous serodetection of multiple infectious agents in laboratory mouse. J Immunol Methods 2010 Dec; 363(1):51-9.