A Multiplex Assay for Simultaneous Detection of Three Retroviruses in Nonhuman Primates Based on the Luminex xTAG Platform

Abstract

Nonhuman primates (NHPs) are widely used for investigating retroviral pathogenesis.Before experiments are conducted and breeding programs established, the animals must be judged negative for endogenous retroviruses.We developed a multiplex high-throughput detection method using multiplexed Luminex fluorescent microbeads and the xTAG system (MMxTAG).We tested three retroviral DNA detection assays for use in a single tube reaction by taking advantage of the Luminex xTAG platform.The assay showed a high sensitivity that was comparable to qPCR methods.The detection limits approached 1-10 fg/μL of plasmid DNA.We compared the specificity, efficiency and accuracy with ELISA assays and found our assay was superior in all regards.The MMxTAG performed well with clinical specimens and could selectively detect a single virus in the presence of the other two.This MMxTAG procedure can be applied to pathogen monitoring in animals and is present in a high throughput format that has low operating and development costs for protecting monkey contacts against these infectious diseases.