Multiplex binding antibody assay for confirmatory HSV-1/2 serological diagnosis

/ABSTRACT Herpes simplex virus types 1 and 2 (HSV-1/2) are major human viral pathogens that cause one of the most common, debilitating, stigmatized and lifelong infections of humankind with more than 143 million new infections every year and affecting more than 4 billion people around the world and more than 100 million Americans. Determining infection status is challenging, as viral latency makes PCR screening both ineffective and costly, while the inadequate specificity of serological IgG assays has led the CDC to recommend a two-step process that includes high throughput, sensitive but low specificity tests followed by more specific but unscalable and labor-intensive confirmatory assays. One of the two CDC-recommended confirmatory assays is the University of Washington (UW) Virology HSV Western Blot test (HSV-WB). The HSV-WB remains the FDA-recognized gold standard for confirmatory serological diagnosis of HSV-1 and HSV-2 infections and detects serum responses to multiple proteins within a viral lysate. This assay is highly sensitive and specific, but it is laborious, and thus costly and only available in the United States at one reference lab. Recurrent quality control problems with another confirmatory assay, biokit gG2-based membrane test, caused product recalls and lack of availability of the test in commercial reference labs. Therefore, making convenient, one-step, high-quality HSV serological testing more accessible is of compelling interest for national and global public health and a key component for development of a national HSV prevention program. In this project, we propose to create a new, high-throughput, one-step HSV serology test that combined the advantages of current FDA-authorized binding assays with the high-content, multiplex-driven specificity of the UW HSV-WB. Specifically, we propose to develop and clinically validate an HSV serologic multiplex bead-based binding antibody assay (HSV Sero-MBAA) that matches the performance characteristics of the HSV-WB, including high sensitivity, specificity, and differentiation of HSV type, on a scalable, inexpensive platform. HSV Sero-MBAA can be run both in central labs and can be kitted and licensed for future regulatory submissions that would allow it to be run in hundreds of clinical laboratories around the United States and world on already existing instrumentation. Project Number: 1R21AI190539-01A1 | Fiscal Year: 2026 | NIH Institute/Center: National Institute of Allergy and Infectious Diseases (NIAID) | Principal Investigator: Anton Sholukh (+1 co-PI) | Institution: FRED HUTCHINSON CANCER CENTER, SEATTLE, WA | Award Amount: $270,591 | Activity Code: R21 | Study Section: Etiology, Diagnostic, Intervention and Treatment of Infectious Diseases Study Section[EDIT] View on NIH RePORTER: https://reporter.nih.gov/project-details/1R21AI19053901A1