Serological Evaluation of Immunity to the Varicella-Zoster Virus Based on a Novel Competitive Enzyme-Linked Immunosorbent Assay

Abstract

Varicella-zoster virus (VZV) is a highly contagious agent of varicella and herpes zoster. Varicella can be lethal to immunocompromised patients, babies, HIV patients and other adults with impaired immunity. Serological evaluation of immunity to VZV will help determine which individuals are susceptible and evaluate vaccine effectiveness. A collection of 110 monoclonal antibodies (mAbs) were obtained by immunization of mice with membrane proteins or cell-free virus. The mAbs were well characterized, and a competitive sandwich ELISA (capture mAb: 8H6; labelling mAb: 1B11) was established to determine neutralizing antibodies in human serum with reference to the FAMA test. A total of 920 human sera were evaluated. The competitive sandwich ELISA showed a sensitivity of 95.6%, specificity of 99.77% and coincidence of 97.61% compared with the fluorescent-antibody-to-membrane-antigen (FAMA) test. The capture mAb 8H6 was characterized as a specific mAb for VZV ORF9, a membrane-associated tegument protein that interacts with glycoprotein E (gE), glycoprotein B (gB) and glycoprotein C (gC). The labelling mAb 1B11 was characterized as a complement-dependent neutralizing mAb specific for the immune-dominant epitope located on gE, not on other VZV glycoproteins. The established competitive sandwich ELISA could be used as a rapid and high-throughput method for evaluating immunity to VZV.

Improving immunization strategies in patients with inflammatory bowel disease

“…Gastroenterologists need to be aware of the limitations of VZV serology using the commercially available immunoassays when they measure varicella immunity in those immunized. We need to rely more upon the patient’s history of immunization, rather than current commercial ELISA, to evaluate the immunity to varicella [54].…”

High Constitutive Interleukin 10 Level Interferes With the Immune Response to Varicella-Zoster Virus in Elderly Recipients of Live Attenuated Zoster Vaccine

“…Two methods were used to evaluate humoral immunity. One was the fluorescent antibody to membrane antigen (FAMA) assay [12], which yields results that correlate with protection from varicella and is accepted as the gold standard against which other methods to measure protective immunity are compared [13]. The other was the glycoprotein enzymelinked immunosorbent assay (gpELISA), which yields results that do not correlate with protection but is the method most commonly used to assess immunity to VZV [10,11] .…”

Current Methods for the Detection of Antibodies of Varicella-Zoster Virus: A Review

“…In addition to indirect ELISA, some researchers have developed a competitive ELISA and a gE double-antigen sandwich ELISA for VZV antibody detection [ 56 , 57 ]. Firstly, for the competitive ELISA (schematic shown in Figure 2 B), an anti-ORF9 antibody is used to capture VZV particles, and then an HRP-labeled anti-gE antibody is used to compete with serum anti-VZV antibodies for virus binding.…”