listeria: how i learned to stop worrying and love the ......listeria monocytogenes was chosen for...
TRANSCRIPT
![Page 1: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/1.jpg)
Brendan Jackson, MD, MPHLCDR US Public Health Service
Division of Foodborne, Waterborne, and Environmental DiseasesCenters for Disease Control and Prevention
National Center for Emerging and Zoonotic Infectious DiseasesDivision of Foodborne, Waterborne, and Environmental Diseases
Listeria: How I Learned to Stop Worrying and Love the Sequencer
With apologies to Mr. Kubrick
![Page 2: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/2.jpg)
Do Cases Represent an Outbreak?
![Page 3: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/3.jpg)
Find Related Cases byPulsed-Field Gel Electrophoresis (PFGE)
=
![Page 4: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/4.jpg)
Limitation: Genetically Unrelated Isolate Might Appear Same by PFGE
=?
![Page 5: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/5.jpg)
Limitation: Genetically Related Isolate Might Appear Different By PFGE
=
?
![Page 6: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/6.jpg)
Listeria WGS Pilot Project
Started September 2013 Goal: Sequence all Listeria monocytogenes isolates Near real-time (<1 week for patient isolates)
Public Health Agency of Canada
National Center for Biotechnology Information
![Page 7: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/7.jpg)
Why Listeria monocytogenes? Illness is rare but serious, costly,
and commonly outbreak associated
Current subtyping methods (e.g., pulsed-field gel electrophoresis) are not ideal
Listeria genome is fairly small and relatively easy to sequence and analyze
Strong epidemiologic surveillance (Listeria Initiative)
Strong regulatory component
![Page 8: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/8.jpg)
Listeria WGS Analysis
Kmer
High-quality single nucleotide polymorphism (hqSNP)
Whole-genome multilocussequence typing (wgMLST)
![Page 9: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/9.jpg)
SNPs wgMLSTallele
differences
1 1
gene 1 gene 2 gene 3
Relationship Between SNPs and wgMLST Alleles
![Page 10: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/10.jpg)
SNPs wgMLSTallele
differences
1 1
2 2
gene 1 gene 2 gene 3
Relationship Between SNPs and wgMLST Alleles
![Page 11: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/11.jpg)
SNPs wgMLSTallele
differences
1 1
2 2
5 2
gene 1 gene 2 gene 3
Relationship Between SNPs and wgMLST Alleles
![Page 12: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/12.jpg)
Listeria Cluster Metrics Before and After WGS
14
0 2 613
19
6 5 4
2021
6 93
98
0
20
40
60
80
100
120
No. of clustersdetected
No. of clustersdetected sooneror only by WGS
No. of outbreakssolved
(food sourceidentified)
Median no. ofcases per
cluster
No. of caseslinked to food
source
Pre-WGS (Sept 2012–Aug 2013)
WGS Year 1 (Sept 2013–Aug 2014)
WGS Year 2 (Sept 2014–Aug 2015)
![Page 13: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/13.jpg)
How WGS Influenced Investigations Show that apparent PFGE clusters are not single-source
outbreaks or are pseudo-clusters
Refine outbreak case definitions by excluding cases
Show that isolates with different PFGE patterns are related
Increase confidence in the link between human and product isolates
Characterize the ecology of long-term pathogen reservoirs in the food chain
![Page 14: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/14.jpg)
BREAKING UP PFGE CLUSTERS
![Page 15: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/15.jpg)
PFGE Cluster Possibly Associated with Uncommon Deli Meats, 2013
0
1
2
3
Week of Isolation
Num
ber o
f Iso
late
s
92% (12/13)reported eating deli meat
(incl. liverwurst, head cheese)
![Page 16: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/16.jpg)
WGS for PFGE Cluster
wgMLST (<All Characters>)
100
959085807570 Id
PNUSAL000156PNUSAL000250PNUSAL000200PNUSAL000146PNUSAL000428PNUSAL000079PNUSAL000029PNUSAL000309PNUSAL000350
63[50-73]
46[26-59]
242[26-274]
251[26-525]
240[26-525]
Allele median[min-max] differences
![Page 17: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/17.jpg)
wgMLST (<All Characters>)
100
959085807570 Id
PNUSAL000156PNUSAL000250PNUSAL000200PNUSAL000146PNUSAL000428PNUSAL000079PNUSAL000029PNUSAL000309PNUSAL000350
63[50-73]
46[26-59]
242[26-274]
251[26-525]
240[26-525]
Allele median[min-max] differences
CLUSTER
![Page 18: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/18.jpg)
REFINING CASE DEFINITIONS
![Page 19: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/19.jpg)
2013 Outbreak Linked to French-Style Soft Cheese
![Page 20: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/20.jpg)
WGS Refines Case Definition (Same PFGE)“How Close is Close?”
WGS analysis by Enteric Diseases Laboratory Branch, CDC
2013L-55432013L-5598
2013L-56702013L-57472013L-5743
2013L-53742013L-5418
2013L-51212013L-5449
2013L-55352013L-5687
2013L-5688
Median: 80SNPs
Median: 9.5 SNPs
Duplicate isolates All isolates shown part of
a PFGE-defined Listeriacluster investigation
Varying levels of relatedness within the PFGE cluster
Can fine-tune analyses by focusing on cases at varying levels of genetic relatedness
Median: 33 SNPs
![Page 21: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/21.jpg)
Epidemiology and Exposure Data Still Needed
WGS analysis by Enteric Diseases Laboratory Branch, CDC
2013L-55432013L-5598
2013L-56702013L-57472013L-5743
2013L-53742013L-5418
2013L-51212013L-5449
2013L-55352013L-5687
2013L-5688
Median: 80SNPs
Median: 9.5 SNPs
Duplicate isolates
Median: 33 SNPsNone of these patients had eaten kale
Isolate from kale
Isolate from kale fairly closely related to other isolates
None of the patients had eaten kale, however
Although more specific, WGS data still need to be interpreted in the context of all the evidence
![Page 22: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/22.jpg)
TYING PFGE PATTERNS TOGETHER
![Page 23: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/23.jpg)
Identified Listeriosis Cluster Not Evident by PFGE
5 cases, 3 PFGE patterns, 3 states All patients originally from
Eastern Europe
8 hqSNPs13.5 hqSNPs
2013 isolate
2 hqSNPs
68 hqSNPsNearest neighbor
WGS analysis by Enteric Diseases Laboratory Branch, CDC
State 1 isolateState 2 isolateState 1 isolate
State 3 isolateState 1 isolate
State 1State 1State 1State 2State 3
![Page 24: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/24.jpg)
Examples from Outbreaks Linked to Ice Cream and Middle Eastern-Style Soft Cheese
wgMLST (<All Characters>)
100
9998
cdc_id
2015L-62872015L-66282014L-63882014L-64172015L-64342013L-55772013L-57182014L-66322013L-54222013L-57482013L-51862015L-61032015L-63282015L-61232014L-66082013L-53462012L-55102013L-51532014L-61822012L-51912010L-21562014L-62392015L-64582015L-65872013L-5195
SourceSite
BLOODBloodBloodBloodBloodBloodBloodBLOODBloodBloodCSFBloodBloodStoolFluidBloodParacentesis BloodBloodBloodBlood, NOSBloodBloodBloodCSFQueso Fresco
14[4-26]
15[4-39]
15[4-101]
wg M LS T (< A ll C ha ra c te rs > )1
00
90
80
70
60
50
40
30
20
10
12.5[0-18]
2571
Allele median[min-max] differences at node
PFG E-AscI-pattern
G X6A16.0026G X6A16.0026G X6A16.0061G X6A16.0020G X6A16.0282
PFG E-ApaI-pa ttern
G X6A12.0227G X6A12.0227G X6A12.0026G X6A12.0227G X6A12.0355
Ice Cream
Soft Cheese
![Page 25: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/25.jpg)
LINK BETWEEN HUMAN AND PRODUCT ISOLATES
![Page 26: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/26.jpg)
0
1
2
3
4
1 8 15 22 29 6 13 20 27 3 10 17 24 31
June July August
Num
ber o
f Cas
es
Week of isolation in 2014
States:AL, IL (4), MA, TN
Cluster Epi
• 6/7 (86%) male • Ages 40-85 years• Two deaths• LI forms for 2 patients • Both ate queso fresco• Matching cheese and
environmental isolates noted from Indiana from 2011
July 2014: PFGE Cluster of Listeriosis Found by PulseNet
![Page 27: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/27.jpg)
hqSNP PhylogenyAug 13, 2014
CA__M14X00709|02/27/2014|2014L-6114
IN__14REF0128|11/17/2013|2013L-5692
IL__131878|09/20/2013|2013L-5579
IL__132446|11/02/2013|2013L-5672
IL__125821|06/12/2013|2013L-5282
IL__134857|07/11/2014|2014L-6385
MA_14EN4302|06/15/2014|2014L-6389
CT__339767001|08/28/2013|2013L-5500
MA_13EN6104|08/06/2013|2013L-5430
MA_13EN7591|missing|2013L-5572
CT__322656001|07/04/2013|2013L-5352
PA__M13016330001A|09/09/2013|2013L-5517
PA__M13017889001A|10/10/2013|2013L-5564
IL__124338|03/28/2013|2013L-5149
IL__134808|06/18/2014|2014L-6325
IL__134856|07/11/2014|2014L-6384
IL__134803|06/07/2014|2014L-6304
IL__124895|04/23/2013|2013L-5175
0.02
1a
Isolate before June 2014 (GX6A16.0319|GX6A12.0246)Cluster 1407MLGX6-1 (GX6A16.0319|GX6A12.0246)
Isolate before June 2014 (GX6A16.0319|GX6A12.0244)
Phylogeny by EDLB
38 [29-47]
46.5 [34-58]
74
2.5 [0-7]
36
New England
clade
Excluded clade
(mostly IL)
PA clade
![Page 28: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/28.jpg)
0
1
2
3
4
1 8 15 22 29 6 13 20 27 3 10 17 24 31
June July August
Num
ber o
f Cas
es
Week of isolation in 2014
Highly-related by WGS
Less related by WGS
Cluster Epidemic Curve
IL ILIL IL MI
![Page 29: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/29.jpg)
Counties where cases are located
Suspicious, but running out of leads
![Page 30: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/30.jpg)
September 4: A Crack in the Case
Isolates uploaded with matching PFGE pattern from mung bean sprouts and sprout irrigation water
Collected as part of routine FDA inspection
Where was the facility?
![Page 31: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/31.jpg)
Exposure Data
LI forms 1 Illinois patient ate sprouts Other patient interviewed with version of questionnaire that didn’t
include sprouts; now unreachable 1/1 sprout consumption vs. 35/760 (5%) historical
cases OR = 20.1 (95% CI, 1.06–infinity) But one case?
![Page 32: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/32.jpg)
Information from Michigan
Patient denies sprout consumption, but reports eating pad thai at several Asian-style restaurants
At least two serve mung bean sprouts
![Page 33: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/33.jpg)
87[87-87]
5[2-6]5[2-54]
55[2-69]
64[2-191]
Environmental IsolateSprout
Allele Median [min-max]
Whole-genome Multilocus Sequence Typing (wgMLST)
100
999897969594
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
TennesseePennsylvaniaIllinoisIllinois
IllinoisIllinoisPennsylvaniaMassachusettsAlabamaIllinois
TennesseePennsylvania
IllinoisIllinoisEnvironmental IsolateSproutIllinoisIllinois
PennsylvaniaMassachusetts
AlabamaIllinois
0.02
68 hqSNPs
1 ± 1 hqSNPs [0-3]
58 hqSNPs [0-72]65.5 hqSNPs [54-72]
275 hqSNPs
WGS analysis by Enteric Diseases Laboratory Branch, CDC
hqSNP Analysis
Patient Isolates Virtually Identical to Sprout Isolates by WGS
Isolate from MI also highly-related (not
shown)
![Page 34: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/34.jpg)
Listeria monocytogenes Outbreak Linked to Mung Bean Sprouts
Despite limited epi data (2/2 consuming sprouts), WGS results and inspection findings enough to take action
CDC and states recommended that consumers not eat and retailers not buy sprouts from the producer
Ultimately the company agreed to permanently cease operations
Federal government later filed lawsuit to prevent firm from resuming production
![Page 35: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/35.jpg)
Caution: Isolates in an Outbreak Are Not Always Highly Related
100
9590858075706560555045
4 [0–13]
5 [0–14]
1563 [0–1696]
Allele differences at node: median [min–max]
Patient isolatesApple and environmental isolates
Cluster 1
Cluster 2
Example of polyclonality
WGS analysis by Enteric Diseases Laboratory Branch, CDC
![Page 36: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/36.jpg)
ECOLOGY OF PATHOGEN RESERVOIRS
![Page 37: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/37.jpg)
2013 Outbreak Linked to Farmstead CheeseCFSAN006121|CFSAN006123CFSAN006122CFSAN004369|CFSAN004358
CFSAN004366|CFSAN004352|
CFSAN004362CFSAN0043492013L-5214CFSAN004354CFSAN004376
CFSAN004367CFSAN004359
CFSAN004372CFSAN004375
CFSAN004371CFSAN004370
CFSAN004357CFSAN004364CFSAN004353CFSAN004368CFSAN004348CFSAN004351CFSAN004350
CFSAN004355CFSAN004365
CFSAN004360CFSAN004363CFSAN004373
CFSAN004377CFSAN004374CFSAN004356
2010L-1790CFSAN004361
2013L-52832013L-5275
2013L-53572013L-5554
2013L-52232013L-5337
2013L-5298
Outbreak cases, 2013<10 high quality
SNPs
Environmental and food samples from cheese-making facility, 2010–2013
2013L-5514
Patient isolate, 2010
WGS analysis by Enteric Diseases Laboratory Branch, CDC
Isolates from 2010 virtually identical to isolates from 2013
![Page 38: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/38.jpg)
2013 Outbreak Linked to Mexican-Style Soft Cheese
663 hqSNPs
6 hqSNPs[2-18]
Cheese ACheese ACheese ACheese BCheese BCheese CCheese ACheese DCheese ECheese FClinical isolateClinical isolateClinical isolateClinical isolateClinical isolateClinical isolate2013 isolate
0.1
Nearest neighbor
WGS analysis by Enteric Diseases Laboratory Branch, CDC
![Page 39: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/39.jpg)
2013–2014 Outbreak Linked to Central and South American-Style Soft Cheeses
wgMLST (<All Characters>)
100
Allele median[min-max] differences at node
13[2-22]
7[2-11]
12[4-18]
5
4[6-9]
7[4-10]
SourceSite
Environmental SwabsEnvironmental SwabsBLOODCheeseCheeseplacentaCSFCheeseBloodBloodplacentaEnvironmental SwabsEnvironmental SwabsCSFUrineBlood
Allele median[min-max] differences at node
13[2-22]
7[2-11]
12[4-18]
5
4[6-9]
7[4-10]
Different PFGE pattern from others
WGS analysis by Enteric Diseases Laboratory Branch, CDC
![Page 40: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/40.jpg)
Listeriosis Linked to Recalled Stone Fruits
• July 2014 recall receives extensive media coverage
• Hundreds of inquiries from concerned clinicians and public
• Stone fruit isolates obtained from company
• 4 human isolates in 2014 with PFGE match
• Was it an outbreak?
![Page 41: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/41.jpg)
MMWR. Mar 20, 2015.
![Page 42: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/42.jpg)
Patients with Recurrent Listeria Infections
~70 year old man: 3 positive blood cultures,attributed to pacemaker lead infection
June 2013 Sept 2013 May 2014
![Page 43: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/43.jpg)
Patients with Recurrent Listeria Infections
~70 year old man: 3 positive blood cultures,attributed to pacemaker lead infection
June 2013 Sept 2013 May 2014
3 hqSNPs
All food removed from home
![Page 44: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/44.jpg)
Patients with Recurrent Listeria Infections
~70 year old man: 3 positive blood cultures,attributed to pacemaker lead infection
June 2013 Sept 2013 May 2014
3 hqSNPs 22 hqSNPs
23 hqSNPs
All food removed from home
![Page 45: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/45.jpg)
Patients with Recurrent Listeria Infections
~70 year old man: 3 positive blood cultures,attributed to pacemaker lead infection
~80 year old man: 2 positive blood cultures 14 months apart; isolates 19 hqSNPs different
June 2013 Sept 2013 May 2014
3 hqSNPs 22 hqSNPs
23 hqSNPs
All food removed from home
![Page 46: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/46.jpg)
Listeria WGS Discussion
Helped detect more clusters and solve more outbreaks
Focused epidemiological resources
Identified new food sources of listeriosis
Epidemiologic data remain essential
WGS not just for Listeria anymore
![Page 47: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/47.jpg)
For more information please contact Centers for Disease Control and Prevention
1600 Clifton Road NE, Atlanta, GA 30333Telephone: 1-800-CDC-INFO (232-4636)Visit: www.cdc.gov | Contact CDC at: 1-800-CDC-INFO or www.cdc.gov/info
The findings and conclusions in this report are those of the authors and do not necessarily represent the official position of the Centers for Disease Control and Prevention.
National Center for Emerging and Zoonotic Infectious DiseasesDivision of Foodborne, Waterborne, and Environmental Diseases
State, Local, and Territorial Health Departments
Advanced Molecular Detection Initiative
Food and Drug Administration
US Department of Agriculture’s Food Safety and Inspection Service
Enteric Diseases Laboratory Branch
Enteric Diseases Epidemiology Branch
Outbreak Response and Prevention Branch
Epidemic Intelligence Service
Atlanta Research and Education Foundation
Acknowledgments
![Page 48: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/48.jpg)
An Early Use of WGS for ListeriaJune 27–July 4, 2013
Day 0Minnesota Dept. of Health notes 2 PFGE-matched
isolates
Day 0PulseNet query identifies 3 more cases in other
states
Day 0All 5 patients report
soft cheeses on Listeria Initiative form
OR = 10.8 (1.8–∞)
Day 13 patients
re-interviewed; 2 name Company A cheese
Day 1 (Friday)Matching isolates found
previously from Company A plant; inspection begins
Day 7 (4th of July)Company A and FDA
announce recall
Day 6 Cheese prelim.
positive for Lm; public warning issued
![Page 49: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/49.jpg)
Pre-Packaged Salad Recall, 2014WGS Links Sporadic Case
March 13: Lettuce recall based on Listeria testing in Canada
April 21: OH inquires about PFGE pattern of lettuce isolate Finds matching case with onset
in late March 6th most common pattern in US
April 25: State learns that patient consumed multiple types of pre-packaged lettuce, including recalled brand
May 13: Canada reports clinical and lettuce isolates have 0 SNP difference among ~2.7 million base pairs examined
![Page 50: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/50.jpg)
100
9998
CN lettuce
WGS Analysis by EDLB, CDC
66 hqSNPs [46-84]
55 hqSNPs [5-66]
24 hqSNPs [0-94]
43 hqSNPs
21.5 hqSNPs [0-33]
58 hqSNPs [46-58]
2013L-5439
2013L-5527
2013L-5545
2013L-5556
2013L-5203
2013L-5204
2013L-5308
2013L-5473
2013L-5085
2013L-5587
2013L-5333
2013L-5314
2013L-5182
2013L-5622
2013L-5623
2013L-5521
2013L-5345
2014L-6017
2013L-5548
Lettuce isolateClinical isolate
2013L-5540
2013L-5359
2013L-5297
2013L-5194
2013L-5560
2014L-6086 0.02
5 hqSNPs
wgMLSThqSNP
![Page 51: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/51.jpg)
An Analogy Attempt
Your OutbreakManuscript
(5,000 pages)
Your manuscript draft returns with edits…
![Page 52: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/52.jpg)
An Analogy Attempt
Your manuscript draft returns with edits…
Lots Your OutbreakManuscript
(5,000 pages)
![Page 53: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/53.jpg)
An Analogy Attempt
Your manuscript draft returns with edits…
Lots
From all 25 coauthors
Your OutbreakManuscript
(5,000 pages)
![Page 54: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/54.jpg)
An Analogy Attempt
Your manuscript draft returns with edits…
Lots
From all 25 coauthors (Don’t these CDC people have
anything better to do?)
Your OutbreakManuscript
(5,000 pages)
![Page 55: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/55.jpg)
An Analogy Attempt
Your manuscript draft returns with edits…
Lots
From all 25 coauthors (Don’t these CDC people have
anything better to do?)
No Track Changes and some of the pages are out of order
Your OutbreakManuscript
(5,000 pages)
![Page 56: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/56.jpg)
How Can You Quantify the Changes?
Kmer Same-length confetti
Your OutbreakManuscript
(5,000 pages)
![Page 57: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/57.jpg)
Kmer Same-length confetti
SNP Every letter
(SNP)
Your OutbreakManuscript
(5,000 pages)
CAT CAT
How Can You Quantify the Changes?
![Page 58: Listeria: How I Learned to Stop Worrying and Love the ......Listeria monocytogenes was chosen for this project because it is rare but serious, costly, and commonly outbreak associated,](https://reader030.vdocuments.mx/reader030/viewer/2022040403/5e8db520949075293679c949/html5/thumbnails/58.jpg)
Kmer Same-length confetti
SNP Every letter
(SNP)
wgMLST Every section
(gene)
Your OutbreakManuscript
(5,000 pages)
How Can You Quantify the Changes?