Technology Bundle ID: TAB-3314

Chimeric Reporter West Nile/Dengue Viruses and Their Use for Assay Development

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Primary Inventors: 
Claire Kinney (CDC)
Development Stage: 
Pre-Clinical (in vitro)
Institute or Center: 
CDC

CDC researchers have engineered West Nile/dengue virus (WN/DENV) chimeras utilizing the replicative ability of the West Nile (WN) virus but presenting the immunogenic pre-membrane and envelope surface proteins of each of the four dengue virus serotypes (DENV 1-4). When coupled with a fluorescent reporter gene, each chimera is able to generate live chimeric reporter WN/DENV (R-WN/DENV) expressing the fluorescent protein in infected cells. These chimeric reporter viruses (CRVs) are used to develop faster and less hands-on high throughput neutralization assays for DENV. In addition, the same CRV platform was also used to develop a chimeric reporter-West Nile/Zika virus (R-WN/ZIKV) (see related technologies below) for the ZIKV neutralization assay. Using a robust WNV replicative vector combined with a reporter that emits a very strong signal, this platform permits live-image detection and measurement of infectivity in 24-30 hours after cell infection by these reporter viruses. A high throughput neutralization assay without labor intensive procedures (such as cell overlay, cell fixation, and/or immunostaining) has been successfully developed for all these 5 CRVs using a live-image based plate reader. With similar replication kinetics for these CRVs, the test protocols can be synchronized for all of these 5 viruses, which is advantageous for samples that may need to be tested against more than one of these viruses for confirmative serologic diagnosis.

Applications:
  • Diagnostic assays for antibody detection of all 4 serotypes of dengue virus
  • High throughput, rapid neutralization assays
  • Same chimeric reporter virus platform for dengue and Zika viruses, and likely for other flaviviruses
  • Antiviral screening and virology research (infection, replication, and pathogenesis)
  • Vaccine candidate immunogenicity testing and validation
  • Monitoring and public health surveillance studies
Advantages:
  • Offers high throughput (scale), easy-to-use procedures and a potentially more sensitive assay
  • Allows fast neutralizing antibody detection in as little as 24-30 hours post-infection
  • Removes the requirement of cellulose or agar overlay, cell fixation, and virus foci/plaque staining in assays
  • The reporter chimeric WN/DENVs replicate faster and more robustly than the whole DENVs
  • The assay developed with these reporter viruses can be directly measured from live cell cultures (live imaging by image-based cytometers or plate readers)
  • Easy and high yield production of the chimeric reporter viruses
  • Genetic and cytometry assays available to confirm the integrity of the reporter gene in the generated CRV stocks

Patents

PCT Application PCT/US2018/046909
Filed on 2018-08-17
US Application 16/639,652
Filed on 2020-02-17
US Application 62/547,527
Filed on 2017-08-18

Updated

Sep 10, 2018

Data Source: 
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