CVI Accepts, published online ahead of print on 7 January 2009

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Clin. Vaccine Immunol. doi:10.1128/CVI.00350-08
Copyright (c) 2009, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Serological Diagnosis of Human HSV-1 and HSV-2 Infection by Luciferase Immunoprecipitation Systems (LIPS) Assay

Peter D. Burbelo, Yo Hoshino, Hannah Leahy, Tammy Krogmann, Ronald L. Hornung, Michael J. Iadarola, and Jeffrey I. Cohen^*

Neurobiology and Pain Therapeutics Section, Laboratory of Sensory Biology, National Institute of Dental and Craniofacial Research, Medical Virology Section, Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Disease, National Institutes of Health, Bethesda, MD 20892; and Clinical Services Program, SAIC-Frederick, Inc., NCI-Frederick, Frederick, Maryland 21702

^* To whom correspondence should be addressed. Email: jcohen{at}niad.nih.gov.

Highly quantitative and high-throughput serological tests to evaluate humoral responses to herpes simplex virus (HSV)-1 and HSV-2 are not available. The efficacy of luciferase immunoprecipitation systems (LIPS) assays for antibody profiling and serologic diagnosis of HSV-1 and HSV-2 infection was investigated using a panel of 5 recombinant HSV antigens. Plasma from subjects seropositive for HSV-1 and/or HSV-2 or seronegative for HSV-1 and HSV-2 that had been previously analyzed by Western blot and Focus Plexus Immunoassay were evaluated. The LIPS test measuring anti-gG1 antibody titers was 96% sensitive and 96% specific for detecting HSV-1 infection compared with the Focus Immunoassay and was 92% sensitive and 96% specific compared with Western blotting. Results from the anti-gG2 LIPS test for HSV-2 precisely matched Western blotting with 100% sensitivity and 100% specificity and showed robust antibody titers in all the HSV-2 infected samples that were over 1,000 times higher than HSV-2 negative or HSV-1 positive samples. Antibodies to 3 additional HSV-2 proteins, gB, gD and ICP8 were detected in many of the HSV-1 and/or HSV-2-infected plasma and showed preferentially higher immunoreactivity in HSV-2 infected plasma. The antibody titers to these 3 HSV-2 antigens also significantly correlated with each other (R=0.75 to 0.81; p<0.0001). These studies indicate that the robust anti-gG1 and anti-gG2 antibody responses detected by LIPS assays are useful for HSV-1 and HSV-2 diagnosis and suggest that profiling of antibody responses to a panel of HSV proteins may be useful for characterizing individual humoral responses to infection and for monitoring responses to vaccines.

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