Rapid point-of-care diagnosis of symptomatic and asymptomatic herpes infection

About This Grant

Herpes simplex virus (HSV) is highly contagious and can be transmitted via physical contact. HSV can be diagnosed by detecting the presence of the virus in lesions or the antibodies in the blood. Yet, viral shedding can happen from asymptomatic infections, highlighting the need for early and accurate detection of HSV to prevent transmission. The most common ways to detect HSV are nucleic acid testing of an active infection via qPCR or serological testing of antibody levels in patient serum. However, qPCR is only accurate if a person is symptomatic and in asymptomatic people both the FDA and the CDC recommend against serological testing due to issues with sensitivity. Additionally, current testing for CNS complications arising from HSV infections requires highly invasive cerebral spinal fluid (CSF) sampling to diagnose. Thus, rapid, accessible, sensitive, and accurate point- of-care tests are in dire need. In 2021, we published a watershed paper describing how we can leverage cell surface glycans that the SARS- CoV-2 virus uses to bind and infect cells, to capture it onto rapid test strips for sensitive detection of the virus (Kim et al, ACS Central Science). Inspired and motivated by our success with SARS-CoV-2 sensing, we propose a novel lateral flow strip assay (LFSA) device for rapid and reliable point-of-care antigen-based detection capable of differentiating between HSV-1 and HSV-2 infections and sensing of CNS complications through serum. As cell surface proteoglycans such as heparan sulfate play an important role in the binding and cell entry of HSV, we will leverage it as a universal binder and use type specific cell receptors to distinguish between HSV strains (Specific Aim 1). Higher selectivity will be achieved by exploring sensitivity to sulfonation of heparan sulfate and other glycocalyx proteins. Sensor performance will be evaluated in complex fluids such as human genital fluids or saliva, and in genital washings of HSV-infected mice. To enhance our ability to identify and subtype HSV, we will engineer tailored cell membranes to optimize their interactions with viral envelope proteins, and strip and print these cell-derived membranes on paper test strips in Specific Aim 2. In Specific Aim 3, we will develop a blood test for the rapid quantitative screening of glial fibrillary acidic protein (GFAP) and neurofilament light chain (NfL), upregulated biomarkers upon CNS damage. We will incorporate electrochemical signals for quantitative assessment. This Bluetooth device will enable early and fast triage of patients for further screening. Together, these devices will enable us rapid and cost-effective screening of high-risk populations, accurate subtyping, and a swift connection of patients with treatment.