Background:
HSV-1 and HSV-2 are common human pathogens that cause a variety of clinical manifestations. Adequate identification of HSV type-specific IgG is important for patient management and sero-epidemiological studies, and recently type-specific HSV-1/2 and ELISA assays have become available. A multiplex assay with equivalent sensitivity and specificity is highly desirable. Here we describe the development of a multiplex bead-based assay using Luminex xMAP technology (Austin, TX) for HSV type-specific antibody detection.

Objective:
Determine the accuracy of a Luminex-based HSV type-specific serology assay versus the ELISA

Method:
Samples: A panel consisting of 158 randomly submitted sera for HSV serology and 28 sera positive for CMV, EBV, ANA, HHV6 and rubella were tested.
HSV-1/2 multiplex assay: Luminex bead sets were covalently linked with specific HSV-1 and -2 recombinant antigens (gG1 and gG2), purified whole HSV-1 and -2 antigens, and a goat-anti-human IgG serving as an internal control. Phycoerythrin (PE) conjugated anti-human IgG was used to detect bound IgG. The amount of specific antibodies bound to the respective beads was expressed in MFIs. The HSV-1 and -2 type-specific ELISAs (HerpeSelect) were performed as described by the manufacturer (Focus Diagnostics Inc., CA). An HSV-1and-2 specific inhibition test was considered as the reference method for discrepant samples.


Result:
The sensitivity of the HSV-1/2 specific multiplex bead-based assay for HSV-1 and -2 compared to ELISA was 98.7% and 100%, respectively, with specificity of 98.8% and 99.1% respectively. None of the 28 cross-reactive samples were positive in the HSV-1/2 multiplex bead-based assay. The correlation between the two assay systems in assigning HSV type-specific serostatus to the sera tested was excellent (98.7%).

Conclusion:
The multiplex bead based immunoassays is a very sensitive and specific method for the combined detection of HSV-1/2 antibodies in serum.

Implications:
The Luminex HSV type-specific assay is a capable replacement for ELISA.