Simultaneous detection of antibodies to five simian viruses in nonhuman primates using recombinant viral protein based multiplex microbead immunoassays.

Routine screening for infectious agents is critical in establishing and maintaining specific pathogen free (SPF) nonhuman primate (NHP) colonies. More efficient, higher throughput, less costly reagent, and reduced sample consumption multiplex microbead immunoassays (MMIAs) using purified viral lysates have been developed previously to address some disadvantages of the traditional individual enzyme-linked immunosorbent assay (ELISA) methods. To overcome some of the technical and biosafety difficulties in preparing antigens from live viruses for viral lysate protein based MMIAs, novel MMIAs using recombinant glycoprotein D precursor (gD) protein of herpesvirus B and four viral gag proteins of simian immunodeficiency virus (SIV), simian T Cell lymphotropic virus (STLV), simian foamy virus (SFV), and simian betaretrovirus (SRV) as antigens have been developed in the current study. The data showed that the recombinant viral protein based MMIAs detected simultaneously antibodies to each of these five viruses with high sensitivity and specificity, and correlated well with viral lysate based MMIAs. Therefore, recombinant viral protein based MMIA is an effective and efficient routine screening method to determine the infection status of nonhuman primates.