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


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.

Potential Commercial Applications: Competitive Advantages:
  • 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
 
  • 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


Related Invention(s):
E-168-2014-0
E-177-2016-0
E-178-2016-0


Inventors:

Claire Kinney (CDC)  ➽ more inventions...


Intellectual Property:
US Application No. 62/547,527
PCT Application No. PCT/US2018/046909

Collaboration Opportunity:

The CDC Technology Transfer Office (TTO) is seeking statements of capability or interest from parties interested in collaborative research to further develop, evaluate, or commercialize: Chimeric Reporter West Nile/Dengue Viruses and Their Use for Assay Development. For collaboration opportunities, please contact CDC TTO at tto@cdc.gov or 1-404-639-1330.


Licensing Contact:
Jeremiah Mitzelfelt,
Email: jeremiah.mitzelfelt@nih.gov
Phone: 301-443-8518

OTT Reference No: E-126-2017-0
Updated: Sep 10, 2018