Loading presentation...

Present Remotely

Send the link below via email or IM

Copy

Present to your audience

Start remote presentation

  • Invited audience members will follow you as you navigate and present
  • People invited to a presentation do not need a Prezi account
  • This link expires 10 minutes after you close the presentation
  • A maximum of 30 users can follow your presentation
  • Learn more about this feature in our knowledge base article

Do you really want to delete this prezi?

Neither you, nor the coeditors you shared it with will be able to recover it again.

DeleteCancel

Visualizing real-time influenza virus infection, transmission

No description
by

Kevin Popham

on 12 November 2015

Comments (0)

Please log in to add your comment.

Report abuse

Transcript of Visualizing real-time influenza virus infection, transmission

CA/09-PA NLuc
Reverse Genetics
NanoLuc
Animal Experiements
Sites of Infection
Patterns of Disease
Background
Methods
Results
Visualizing real-time influenza virus infection, transmission and protection in ferrets

Paper by:
Erik A. Karlsson, Victoria A. Meliopoulos, Chandra Savage, Brandi Livingston, Andrew Mehle & Stacey Schultz-Cherry
Kevin Popham & Thomas Gentile
Image courtesy of Promega
Modified deep sea shrimp luciferase
100 times more intense than from firefly and
Renilla
luciferases
NanoLuc gene was cloned onto the influenza polymerase subunit gene (PA)
Used a viral isolate from the 2009 H1N1 pandemic
Image courtesy of Smith/Science
Biological Applications
Eyes/Conjunctiva
Ears
Kidneys
Lower Respiratory Tract
Upper Respiratory Tract
Nose
Animals that were infected through direct contact had a greater diversity of tissue trophism
Traditional nasal titres are unable to detect infection in some cases
Site of primary infection may impact adaptive immunity
Seasonal Flu
Measure the dynamics of this year's flu strain
Help develop effective public health interventions
Test the efficacy of vaccine candidates
Further Applications
Created a new viral strain called A/California/04/2009 NLuc (CA/09-PA NLuc)
No loss of replicative abilities compared to parent strain
Study virulent influenza strains like the Spanish and avian flu
Study novel emerging and zoonotic diseases
Detect subclinical infections
Fluorescence-based microneutralization assays
Post Reverse Genetics
In Vitro Infections with MDCK Cell Line
Aliquots of infected culture
supernatant were collected
periodically post infection,
stored at -80C for determination
of virus titres.
Luminescent TCID50 Assays
Assay optimized to be read within 18hr after titrating virus on MDCK as compared to 72hr for traditional TCID50 Assay
Bioluminescent Imaging with Xenogen IVIS200 System + LivingImage Software
Nasal washes collected in a specimen cup, viral titres determined by TCID50 analysis.
Full transcript