Abstract
Foodbone norovirus (NoV) is the leading cause of acute gastroenteritis worldwide. Candidate vaccines are being developed, however, no licensed vaccines are currently available for managing NoV infections. Screening for stimulated antibodies with broad-spectrum binding activities can be performed for the development of NoV polyvalent vaccines. In this study, we aimed to develop an indirect enzyme-linked immunosorbent assay (ELISA) for testing the broad spectrum of anti-NoV antibodies. Capsid P proteins from 28 representative NoV strains (GI.1–GI.9 and GII.1–GII.22 except GII.11, GII.18, and GII.19) were selected, prepared, and used as coating antigens on one microplate. Combined with incubation and the horseradish peroxidase chromogenic reaction, the entire process for testing the spectrum of unknown antibodies required 2 h for completion. The intra-assay and inter-assay coefficients of variation were less than 10%. The new method was successfully performed with monoclonal antibodies and polyclonal antibodies induced by multiple antigens. In conclusion, the indirect ELISA assay developed in this study had a good performance of reliability, convenience, and high-throughput screening for broad-spectrum antibodies.
Highlights
Foodbone Norovirus (NoV) is one of the leading causes of epidemic acute gastroenteritis (AGE) worldwide, accounting for one-fifth of all gastroenteritis cases globally (Lopman et al, 2016)
glutathione S-transferase (GST) bound with GST monoclonal antibodies at optical density (OD) values of 0.98, 0.96, and 0.98, indicating that the developed enzyme-linked immunosorbent assay (ELISA) technique was reliable
Owing to the high frequencies of NoV outbreaks (Hall et al, 2011; Verhoef et al, 2013; Grytdal et al, 2016) and severe acute gastroenteritis (Atmar and Mullen, 2013; Cardemil et al, 2017), high risk of infection transmission (Sukhrie et al, 2010; Teunis et al, 2010; Verhoef et al, 2015), and enormous socioeconomic burden after infection (Pires et al, 2015), the development of NoV vaccines is critical for improving public health
Summary
Foodbone Norovirus (NoV) is one of the leading causes of epidemic acute gastroenteritis (AGE) worldwide, accounting for one-fifth of all gastroenteritis cases globally (Lopman et al, 2016). NoV has been estimated to cause 677 million episodes of diarrheal disease (95% uncertainty interval [UI]: 468–1,153 million) and 213,515 deaths (95% UI: 171,783–266,561) for all ages and among all modes of transmission (Pires et al, 2015). NoV is extremely contagious, and transmission occurs directly from person to person via fecal-oral and vomit-oral routes, but can be caused by food-borne, water-borne, or environmental factors (Verhoef et al, 2015). Vomitus and feces of infected patients contain a considerable number of virions, whereas as few Testing of NoV Antibody as 10 infectious particles are sufficient to cause AGE (Teunis et al, 2010). Owing to the significant social and economic burden associated with the disease, adequate preventive measures against this virus should be formulated
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