Detection of Antibodies to Varicella-Zoster Virus in Recipients of the Varicella Vaccine by Using a Luciferase Immunoprecipitation System Assay

Abstract

A high-throughput test to detect varicella-zoster virus (VZV) antibodies in varicella vaccine recipients is not currently available. One of the most sensitive tests for detecting VZV antibodies after vaccination is the fluorescent antibody to membrane antigen (FAMA) test. Unfortunately, this test is labor-intensive, somewhat subjective to read, and not commercially available. Therefore, we developed a highly quantitative and high-throughput luciferase immunoprecipitation system (LIPS) assay to detect antibody to VZV glycoprotein E (gE). Tests of children who received the varicella vaccine showed that the gE LIPS assay had 90% sensitivity and 70% specificity, a viral capsid antigen enzyme-linked immunosorbent assay (ELISA) had 67% and 87% specificity, and a glycoprotein ELISA (not commercially available in the United States) had 94% sensitivity and 74% specificity compared with the FAMA test. The rates of antibody detection by the gE LIPS and glycoprotein ELISA were not statistically different. Therefore, the gE LIPS assay may be useful for detecting VZV antibodies in varicella vaccine recipients. (This study has been registered at ClinicalTrials.gov under registration no. NCT00921999.) V aricella-zoster virus (VZV) causes both chickenpox and zoster. A live attenuated varicella vaccine derived from the Oka strain of the virus was developed by Takahashi and colleagues in the 1970s and was licensed for routine use in the United States in 1995. One of the most sensitive tests for detecting VZV antibodies after vaccination is the fluorescent antibody to membrane antigen (FAMA) test. For this test, serial dilutions of human serum are incubated with live VZV-infected human fibroblasts, incubated with fluorescein-tagged anti-human immunoglobulin, and examined by fluorescence microscopy (1). The test detects antibodies to surface glycoproteins on live VZV-infected cells. While the FAMA test is highly predictive of protection from varicella infection after vaccination (2), the test is labor-intensive and somewhat subjective to read. Therefore, the FAMA assay is not applicable for largescale or commercial testing, nor is it readily available.Most laboratories use commercial enzyme-linked immunosorbent assays (ELISAs) to determine VZV seropositivity. A comparison of the commercially available ELISA with the FAMA test in recipients of the varicella vaccine indicates that the ELISA has a sensitivity of 74% and a specificity of 89% (3) (assuming that the FAMA has 100% sensitivity and 100% specificity). Thus, the ELISA is not considered sufficiently sensitive for reliably detecting antibodies after varicella immunization. Several studies have reported failures to seroconvert after immunization even after 2 doses, based on ELISA (4), and these are thought to represent a failure to detect antibody responses rather than a failure of the vaccine.Modified FAMA tests have been developed, including ones that use fixed cells (5) and a flow cytometry-based FAMA assay using virus-infected cells (6). The fixed-cells test is subjective to ...

Herpesvirus Infections and Childhood Arterial Ischemic Stroke

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Antibody Engineering for Pursuing a Healthier Future

“…Rare proteins can be accumulated up to 10,000-fold by IP (Dwane and Kiely, 2011). Luciferase immunoprecipitation systems (LIPS) have been developed for the rapid detection of antibodies against peste des petits ruminants virus (PPRV) (Berguido et al, 2016), varicella-zoster virus (VZV) (Cohen et al, 2014), zinc transporter (ZnT8) autoantibodies (Ustinova et al, 2014), and pancreatic and duodenal homeobox 1 autoantibodies (PAA) (Donelan et al, 2013).…”

Lethal Hemorrhagic Disease and Clinical Illness Associated with Elephant Endotheliotropic Herpesvirus 1 Are Caused by Primary Infection: Implications for the Detection of Diagnostic Proteins

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