Human caliciviruses are among the most important causes of gastroenteritis. Despite the increasing awareness of these agents as a major cause of diarrheal illness, assays for the diagnosis of human calicivirus infection are not available to most clinical laboratories. There are also no effective antivirals available that can be used therapeutically or prophylactically for this infection. This project will focus on high throughput approaches to develop rapid diagnostic assays for the detection of human caliciviruses and high throughput assays to identify potentially effective antiviral agents. We hypothesize that the reagents and strategies developed in the previous project period to target shared epitopes can be optimized and used to develop broadly reactive diagnostic assays for these antigenically diverse viruses.
In Specific Aim 1, we will further optimize broadly-reactive reagents that can be used to make diagnostic assays for human caliciviruses. Studies in this aim will optimize single chain (scFv) antibodies with a range of binding specificities and monoclonal antibodies that recognize conserved epitopes, both of which were developed and characterized in the previous funding period.
In Specific Aim 2, we will develop broadly-reactive diagnostic assays for human caliciviruses. Rapid formats that will be useable in the field to detect human caliciviruses will be identified. High affinity, cross-reactive single chain and monoclonal antibodies characterized in specific aim 1 will be used in assays that require minimal sample preparation and yield a result in less than 30 minutes. Formats to be evaluated are those that have been used successfully for the detection of other human viruses and include solid-phase immunoassays, and latex agglutination assays. In Speciflc Aim 3, we propose to develop high throughput assays to screen agents for antiviral activity against human noroviruses. A high throughput screen for inhibitors ofthe viral protease will be used to identify inhibitor candidates from small molecule libraries. The results obtained from project 1 studies will lead to the availability of new assays for the diagnosis of these important enteric pathogens and the identification of antiviral agents with the potential to diminish the disease burden caused these agents.

Public Health Relevance

Noroviruses are a major cause of diarrheal disease, affecting travelers (for example, those on cruise ships), young children, the elderiy. and other groups. However, there are no readily available tests for rapidly diagnosing infection, and there are no medicines available to treat or prevent such infections. This project will develop rapid diagnostic tests and will identify possible antiviral agents for norovirus infection.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program Projects (P01)
Project #
5P01AI057788-09
Application #
8450095
Study Section
Special Emphasis Panel (ZAI1-GPJ-M)
Project Start
Project End
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
9
Fiscal Year
2013
Total Cost
$307,628
Indirect Cost
$107,219
Name
Baylor College of Medicine
Department
Type
DUNS #
051113330
City
Houston
State
TX
Country
United States
Zip Code
77030
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Qu, Lin; Vongpunsawad, Sompong; Atmar, Robert L et al. (2014) Development of a Gaussia luciferase-based human norovirus protease reporter system: cell type-specific profile of Norwalk virus protease precursors and evaluation of inhibitors. J Virol 88:10312-26
Venkataram Prasad, B V; Air, Gillian M (2014) Editorial overview: virus-glycan interactions and pathogenesis. Curr Opin Virol 7:v-vi
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Katayama, Kazuhiko; Murakami, Kosuke; Sharp, Tyler M et al. (2014) Plasmid-based human norovirus reverse genetics system produces reporter-tagged progeny virus containing infectious genomic RNA. Proc Natl Acad Sci U S A 111:E4043-52
Ramani, Sasirekha; Atmar, Robert L; Estes, Mary K (2014) Epidemiology of human noroviruses and updates on vaccine development. Curr Opin Gastroenterol 30:25-33
Shanker, Sreejesh; Czako, Rita; Sankaran, Banumathi et al. (2014) Structural analysis of determinants of histo-blood group antigen binding specificity in genogroup I noroviruses. J Virol 88:6168-80
Venkataram Prasad, B V; Shanker, Sreejesh; Hu, Liya et al. (2014) Structural basis of glycan interaction in gastroenteric viral pathogens. Curr Opin Virol 7:119-27
Rogers, Jennifer D; Ajami, Nadim J; Fryszczyn, Bartlomiej G et al. (2013) Identification and characterization of a peptide affinity reagent for detection of noroviruses in clinical samples. J Clin Microbiol 51:1803-8

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