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Norovirus Testing: What Happens at “The Lab”

Norovirus Testing: What Happens at “The Lab”. Dr. Amy Woron Amy.Woron@TN.gov 615-262-6462. Molecular Biologist TN Dept. of Health Laboratory Services. Norovirus GI and GII Assay. Differentiates between GI and GII Can have results in as little as 5 hours. 2009 data below. GI. GII.

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Norovirus Testing: What Happens at “The Lab”

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  1. Norovirus Testing:What Happens at “The Lab” Dr. Amy Woron Amy.Woron@TN.gov 615-262-6462 Molecular Biologist TN Dept. of Health Laboratory Services

  2. Norovirus GI and GII Assay • Differentiates between GI and GII • Can have results in as little as 5 hours 2009 data below GI GII

  3. CaliciNet • There are multiple patients all with norovirus GII. • Are they all related? Maybe.

  4. CaliciNet USA • Best tool is the epi investigation ….. • CaliciNet can help National Norovirus Outbreak Network

  5. Current Pitfalls in Norovirus Surveillance No national laboratory surveillance system Tracking multi-state (international) outbreaks with a potential common source is a challenge Only a fraction of the states report to the CDC No standard protocol among laboratories for sequence analysis and genotyping

  6. Objectives of CaliciNet Improve surveillance of norovirus (sapovirus, other GI viruses) Allow for real-time data exchange at the state and national level Rapid genotyping and compare norovirus sequences Link geographically different clusters of illness to a common (food) source Facilitate the identification of new emerging norovirus strains

  7. States Participating in CaliciNet WA WA ME ME ME MT MT ND ND MN MN VT OR OR ID NH MA SD SD NY NY WI WI WY WY RI MI MI CT PA PA IA IA NE NE NJ NV NV OH OH DE MD MD UT UT IN IN CO CO IL IL DC CA CA WV WV VA VA KS KS MO MO KY KY NC NC TN OK OK AZ AZ AR AR SC SC NM NM MS MS AL AL GA GA HI TX TX LA LA AK AK FL FL States with CaliciNet certified participants March 2010 (n = 16)

  8. Confirmed and Suspected Etiology of 1270 Foodborne Outbreaks Reported to CDC, 2006 Other/Multiple 2% Unknown 29% Norovirus 40% Chemical 5% Parasites 1% Bacteria CDC 2009 MMWR 23%

  9. Norovirus testing: From Stool Cup to Sequence RNA Extraction Stool specimen Conventional RT-PCR RT-PCR testing Real-time RT-PCR positive Compare norovirus sequence with database Cycle sequencing

  10. Norovirus regions targeted by detection and typing RT-PCR assays VP2 Hel Pro Pol VP1 7654 1 3' 5' ORF 3 ORF 1 ORF 2 RT* C D 85/98 bp 330/344 bp 177/253 bp * RT = TaqMan realtime RT-PCR Norovirus typing Norovirus detection

  11. Required fields for CN upload Lab Epi Page 8; CN Manual v.3

  12. Multiple Alignment

  13. Dendrogram

  14. Nucleotide Polymorphisms

  15. GI Reference Strains

  16. GII Reference Strains

  17. Similarity to PulseNet • BioNumerics • Encourage state level participation • Increase real-time information flow • Ability to view/compare outbreaks at the national level • Another tool in the epidemiology tool belt

  18. CaliciNet Summary A novel national surveillance system for linking norovirus outbreak clusters potentially to a common source Network for standardized norovirus genotyping and classification Potential to identify new emerging norovirus strains

  19. CDC CaliciNet Team Contact information: Leslie Barclay lbarclay@cdc.gov (CaliciNet scripts) Nicole Gregoricus ngregoricus@cdc.gov (Laboratory protocols) Jan Vinjé jvinje@cdc.gov (Supervisor CaliciNet)

  20. Human Astrovirus testing … Human Sapovirus testing … Food testing for norovirus … Slides with the CDC CaliciNet logo courtesy of Dr. Jan Vinjé, CaliciNet Team Leader, CDC Future Study

  21. Questions?

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