tools - engage benchmarking for amr gene detection · tools - engage benchmarking for amr gene...

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1 Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology, DTU-Food WHO Collaborating Centre for Antimicrobial Resistance in Foodborne Pathogens and Genomics European Union Reference Laboratory for Antimicrobial Resistance (EURL-AMR) The Use of Whole Genome Sequencing (WGS) in Monitoring of Antimicrobial Resistance EURL-AR Training course 2017

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Page 1: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

1

Tools - ENGAGE benchmarking for AMR gene detection

Pimlapas Leekitcharoenphon (Shinny)Research Group of Genomic Epidemiology, DTU-Food

WHO Collaborating Centre for Antimicrobial Resistance in Foodborne Pathogens and Genomics

European Union Reference Laboratory for Antimicrobial Resistance (EURL-AMR)

The Use of Whole Genome Sequencing (WGS) in Monitoring of Antimicrobial Resistance EURL-AR Training course 2017

Page 2: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Page 3: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

3

http://www.engage-europe.eu/resources/protocols-and-training

Page 4: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Page 5: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Page 6: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Page 7: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Page 8: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Page 9: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Page 10: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Page 11: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Page 12: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Global phylogeny of DT104

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Time1960 1970 1980 1990 2000 2010

1.E1

1.E2

1.E3

1.E4

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2.97 x 10-7 SNP/site/year

4619 variable sites

~ 1.47 SNP/year

1946 (1931 - 1959)

109

60 (n = 297)

103

MDR DT104

Leekitcharoenphon P. et al, Appl Environ Microbiol. 2016

Page 13: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Time1960 1970 1980 1990 2000 2010

1.E1

1.E2

1.E3

1.E4

Eff

ectiv

e po

pula

tion

size

Time1960 1970 1980 1990 2000 2010

Demographic history of global DT104

Leekitcharoenphon P. et al, Appl Environ Microbiol. 2016

Page 14: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Page 15: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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http://itol.embl.de

Page 16: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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DTU2013 1369 PRJ941 Vibrio cholerae P47 V cholerae El Tor Ogawa R1.trim.fq.sorted

DT

U2015 512 P

RJ1014 V

ibrio Cholerae C

MC

091 R1

3735-CDC China 1988 1

2011EL-2320 Haiti 2011 R1

V01-CDC Chile 1991 1

DTU

2015

-847

-PR

J101

4-V

ibrio

-cho

lera

e-13

AR

SC

MC

-030

-757

S8

L001

R1

001.

fast

q.so

rted 93284-C

DC

China 1993 1

VC-9 Nepal 2010 R1

CP1040

Zam

bia 20

04 R

1

4339 India 2004 R1

IDH

O1

726

Indi

a 20

09 R

1

DT

U2015 483 P

RJ1014 V

ibrio Cholerae E

VR

003 R1

DT

U2015 516 P

RJ1014 V

ibrio Cholerae G

MH

033 R1

HFU-02 Haiti 2010 R1

1627-CD

C C

hina 1997 1

DTU

2015

506

PR

J101

4 V

ibrio

Cho

lera

e V

SM

718

R1

D118-CDC China 1961 1

DTU2013 933 PRJ923 Vibrio cholera 330 2A R1

DTU

2013 1341 PR

J941 Vibrio cholerae V

cholerae El Tor Inaba 22115 R

1.trim.fq.sorted

VC-4 N

epal 2010 R1

7682

Ken

ya 2

009

R1

A184 Colombia 1992 1

SF20016 Bangladesh 1976

1593

-CDC C

hina

199

2 1

GD196110-CDC China 1961 1

DTU2013 919 PRJ923 Vibrio cholera 068 9A R1

4679

Ban

glad

esh

1999

1

S27R

G11 P

akistan 2010 1

VC22

48-1

-CD

C C

hina

200

8 1

2010EL-1792 Haiti 2010

PR

L64 India 1992 1

HC-68A1 Haiti 2010 R1

DTU

2013 1346 PR

J941 Vibrio cholerae V

cholerae El Tor O

gawa 22126 R

1.trim.fq.sorted

DTU

2013

135

4 PR

J941

Vib

rio c

hole

rae

P37

V ch

oler

ae E

l Tor

Inab

a R

1.tri

m.fq

.sor

ted

4121 Vietnam

2004 R1

VC-13 N

epal 2010 R1

HC-41A1 Haiti 2010 R1

DTU2013

1338

PRJ9

41 V

ibrio

chole

rae 5

01 77

1 R1.t

rim.fq

.sorte

d

A22 Bangladesh 1979 1

DTU

2015

467

PR

J101

4 Vi

brio

Cho

lera

e C

MC

019

R1

DTU2013 915 PRJ923 Vibrio cholera 021 5A R1

DTU2013 923 PRJ923 Vibrio cholera 150 7A R1

DTU

2015

495

PR

J101

4 V

ibrio

Cho

lera

e B

RH

120

R1

HC-42A1 Haiti 2010 R1

2015EL-1158 Kenya unkn 1

A346 1 Bangladesh 1994 1

9-CD

C C

hina 2001 1

HC-46A1 Haiti 2010 R1

4672 Bangladesh 2000 R1

3075-CDC China 1974 1

S26

R24

Pak

ista

n 20

10 1

A10

Bang

lade

sh 1

979

1

DTU2013 1360 PRJ941 Vibrio cholerae P46 V cholerae El Tor Inaba R1.trim.fq.sorted

DTU2013 918 PRJ923 Vibrio cholera 066 13A R1

DTU2013

1337

PRJ9

41 V

ibrio

chole

rae 8

16 ct

634 R

1.trim

.fq.so

rted

DT

U20

15 4

86 P

RJ1

014

Vib

rio C

hole

rae

EV

R 1

21 R

1

A29 Peru 1991 1

1583

-CDC

Chi

na 1

991

1

DTU

2013 1345 PR

J941 Vibrio cholerae V

cholerae El Tor O

gawa 22125 R

1.trim.fq.sorted

2865-CDC China 1961 1

DTU2013 1362 PRJ941 Vibrio cholerae P8 V cholerae El Tor Ogawa R1.trim.fq.sorted

MJ1236 Bangladesh 1994

SF20011 Bangladesh 1969

VC-22 Nepal 2010 R1

1362 Mozam

bique 2005 R1

DTU2013 917 PRJ923 Vibrio cholera 063 9A R1

2011

EL-

1141

Afg

hani

stan

200

8 R

1

NH

CC

-004

A B

angl

ades

h 20

10 R

1

X190-CDC Peru 1991 1

S24R

G6 P

akistan 2010 1

2010EL-1799 Haiti 2010 R1

SF20012 Bangladesh 1969

HC-49A2 Haiti 2010 R1

4646 India 2007 R1

HC-70A1 Haiti 2010 R1

2011EL-2317 Haiti 2011 R1

DTU2013 932 PRJ923 Vibrio cholera 315 4A R1

DTU2013 921 PRJ923 Vibrio cholera 094 13A R1

2015EL-1156 Kenya unkn 1

DT

U20

15 4

88 P

RJ1

014

Vib

rio C

hole

rae

EV

R 1

23 R

1

2010EL-2010N Haiti 2010

CP

1050

Ban

glad

esh

2010

R1

CP1041

Zam

bia 20

04 R

1

GP160 India 1980 1

P27457 Bangladesh 1976

4538 India 2007 R1

S9KCH9 Pakistan 2010 1

7684

Ken

ya 2

009

R1

2308-CDC China 1978 1

6201

Ken

ya 20

07 R

1

DT

U2015 510 P

RJ1014 V

ibrio Cholerae C

MC

089 R1

DTU2013 930 PRJ923 Vibrio cholera 281 8A R1

DTU20

13 1

375

PRJ9

41 V

ibrio

cho

lera

e VC

O1

9 R1.

trim

.fq.s

orte

d

DTU2013 931 PRJ923 Vibrio cholera 285 11A R1

RND6878 Russia 2012

DT

U20

15 5

14 P

RJ1

014

Vib

rio C

hole

rae

GM

H 0

31 R

1

DTU2

013

1355

PRJ

941

Vibr

io c

hole

rae

P38

V ch

oler

ae E

l Tor

Inab

a R1

.trim

.fq.s

orte

d

DTU2013 1365 PRJ941 Vibrio cholerae P33 V cholerae El Tor Ogawa R1.trim.fq.sorted

3497-CDC China 1980 1

A397 Bangladesh 1987 1

3014-CDC China 1969 1

VC-5 Nepal 2010 R1

DTU

2015

479

PR

J101

4 V

ibrio

Cho

lera

e C

MC

031

R1

2744-CDC China 1962 1

HC-20A2 Haiti 2010

2011EL-2318 Haiti 2011 R1

HC-47A1 Haiti 2010 R1

2981-CDC China 1965 1

DTU

2013

136

4 PR

J941

Vib

rio c

hole

rae

P25

V ch

oler

ae E

l Tor

Oga

wa

R1.

trim

.fq.s

orte

d

DTU

2015

504

PR

J101

4 V

ibrio

Cho

lera

e V

SM

716

R1

A232 Mexico 1991 1

DTU2013 1368 PRJ941 Vibrio cholerae P45 V cholerae El Tor Ogawa R1.trim.fq.sorted

A488S

xx2xxE B

angladesh 2006 R1

HC-43A1 Haiti 2010

DT

U20

15 4

85 P

RJ1

014

Vib

rio C

hole

rae

EV

R 1

20 R

1

A18 India 1977 1

C6706-CDC Peru 1991 1

A186 Argentina 1992 1

2752-CDC China 1981 1

RND19191 Russia 2001

DTU2013 935 PRJ923 Vibrio cholera 342 3A R1

HC-06A1 Haiti 2010 R1

4210-CD

C C

hina 1999 1

G4222 South Africa 2001

2011EL-2316 Haiti 2011 R1

DTU

2015

493

PR

J101

4 V

ibrio

Cho

lera

e B

RH

118

R1

Country

Philipines

China

Thailand

Pakistan

Bangladesh

India

Nepal

Vietnam

Indonesia

Afghanistan

Bahrain

Malaysia

Haiti

USA

Mexico

Dominican_Republic

Kenya

Djibouti

Mozambique

South_Africa

Zambia

Mauritania

Tanzania

Zimbabwe

Peru

Argentina

Colombia

Bolivia

Chile

Russia

Cameroon

Continent

Asia

North_America

Africa

South_America

Russia

Biotype

El_Tor

El_Tor_variant

Tree scale: 0.01

S23HH18 Pakistan 2010 1

6191

Keny

a 2005 R

1

DTU2013 916 PRJ923 Vibrio cholera 041 A R1

3626-CDC China 1983 1

2015EL-1157 Kenya unkn 1

HC-77A1 Haiti 2010 R1

DT

U20

15 4

87 P

RJ1

014

Vib

rio C

hole

rae

EV

R 1

22 R

1

143-CD

C C

hina 2001 1

F1D

N4

Pak

ista

n 20

10 1

7685

Ken

ya 2

009

R1

VC-18 Nepal 2

010 R1

1627 Mozam

bique 2005 R1

DTU

2015

499

PR

J101

4 V

ibrio

Cho

lera

e V

SM

657

R1

DT

U20

15 4

91 P

RJ1

014

Vib

rio C

hole

rae

EV

R 2

87 R

1

3289-CDC China 1978 1

F18K

TH

3 Pakistan 201 0 1

DTU2013 913 PRJ923 Vibrio cholera 004 18A R1

VC-17 Nepal 2010 R1

DTU2016-1979-PRJunkn-Vibrio-cholerae-VC-3 S3 L001 R1 001.trim.fq.sorted

A481

Djib

outi

2007

R1

VC

-6 N

epal

201

0 R

1

DTU2013 936 PRJ923 Vibrio cholera 342 10A R1

1909-CD

C C

hina 1998 1

DTU2013 1366 PRJ941 Vibrio cholerae P36 V cholerae El Tor Ogawa R1.trim.fq.sorted

2015EL-1161 Kenya unkn 1

AHV1003-CDC China 2010 1

3304-CDC China 1979 1

DTU

2015

498

PR

J101

4 V

ibrio

Cho

lera

e V

SM

656

R1

SF20014 Bangladesh 1969

3564-CDC China 1982 1

A131 India 1989 1

VC-21113 N

epal 2003 R

1

2011EL-2314H Haiti 2011 R1

L-3226 Russia 2010

3546-06 India 2006 R1

V109 India 1990 1

4642 India 2006 R1

DTU

2015

509

PR

J101

4 V

ibrio

Cho

lera

e V

SM

721

R1

HC-81A1 Haiti 2010

2011EL-1841 Haiti 2011 R1

A241

Viet

nam

198

9 1

HC-28A1 Haiti 2010 R1

S8K

CH

18 Pakistan 2010 1

6310-CDC Indonesia 1961 1

A245

Vietna

m 1

989

1

HC-67A1 Haiti 2010 R1

2012V-1060 USA FL 2012 R1

DTU2013 1370 PRJ941 Vibrio cholerae P48 V cholerae Ei Tor Ogawa R1.trim.fq.sorted

HC-19A1 Haiti 2010 R1

DTU

2013

135

7 PR

J941

Vib

rio c

hole

rae

P42

V ch

oler

ae E

l Tor

Inab

a R

1.tri

m.fq

.sor

ted

F8D

25 Pakistan 2010 1

HC-57A2 Haiti 2010 R1

DTU

2015

475

PR

J101

4 Vi

brio

Cho

lera

e C

MC

027

R1

A316 Argentina 1993 1

4536 India 2007 R1

S17

HH

3 P

akis

tan

2010

1

4784

Tanz

ania

2009

R1

DTU

2013

135

9 PR

J941

Vib

rio c

hole

rae

P44

V ch

oler

ae E

l Tor

Inab

a R

1.tri

m.fq

.sor

ted

DTU2013 929 PRJ923 Vibrio cholera 278 8A R1

C6706 Peru 1991

HC-80A1 Haiti 2010 R1

6193 Kenya

2005 R1

2009V-1096 USA SxxIndiaxxE

2009

M66-2 Indonesia 1937

DTU

2015

496

PR

J101

4 V

ibrio

Cho

lera

e B

RH

121

A R

1

V5 India 1989 1

DTU

2013 1347 PR

J941 Vibrio cholerae V

cholerae El Tor O

gawa 22127 R

1.trim.fq.sorted

VC-21 Nepal 2010 R1

A185 Colombia 1992 1

936-CD

C C

hina 1998 1

DTU

2015

503

PR

J101

4 V

ibrio

Cho

lera

e V

SM

661

R1

2010EL-2026 Haiti 2010 R1

2011EL-1818 Haiti 2011 R1

2894-CDC China 1961 1

3024-CDC China 1973 1

DTU

2014

105

5 PR

J941

Vib

rio c

hole

rae

4053

0243

03 R

1.tri

m.fq

.sor

ted

GP152 India 1979 1

VC-12 N

epal 2010 R1

F4D

48 P

akis

tan

2010

1

4070-CD

C C

hina 1994 1

DTU

2015

471

PR

J101

4 Vi

brio

Cho

lera

e C

MC

023

R1

DT

U2015 515 P

RJ1014 V

ibrio Cholerae G

MH

032 R1

A6 Indonesia 1957 1

4039-CD

C C

hina 1993 1

2011EL-2315 Haiti 2011 R1

DTU

2015

477

PR

J101

4 Vi

brio

Cho

lera

e C

MC

029

R1

2010EL-1787 Haiti 2010 R1

1346 Mozam

bique 2005 R1

DTU2013 938 PRJ923 Vibrio cholera 355 1A R1

VC

-19

Nep

al 2

010

R1

A32 Peru 1991 1

SF20015 Bangladesh 1969 2011EL-2323 Haiti 2011 R1

V212-1 India 1991 1

KS72

0 Ba

ngla

desh

201

1 R

1

F5D

38 P

akis

tan

2010

1

3633-CDC China 1984 1

956-CD

C C

hina 2001 1

2710

-CDC C

hina

1986

1

A200 Argentina 1992 1

NH

CC

-010

F B

angl

ades

h 20

10 R

1

2012EL-2176 Haiti 2012

2010EL-1795 Haiti 2010 R1

4322 India 2004 R1

MJ1485 Bangladesh 1994 1

VC-21106 N

epal 2003 R

1

H1 Haiti 2010

DTU

2013 1348 PR

J941 Vibrio cholerae V

cholerae El Tor O

gawa 22128 R

1.trim.fq.sorted

642345-CDC China 1964 1

VC-7 Nepal 2010 R1

DT

U20

15 5

13 P

RJ1

014

Vib

rio C

hole

rae

GM

H 0

19 R

1

2011V-1021 Dominican Republic 2011

DTU

2015

465

PR

J101

4 Vi

brio

Cho

lera

e C

MC

017

R1

DTU2013 939 PRJ923 Vibrio cholera 322 3E R1

F7D

30 P

akis

tan

2010

1

RND19187 Russia 2010

2011

EL-

1938

Indi

a 20

09 R

1

S12

P76

Pak

ista

n 20

10 1

HC-48B2 Haiti 2010 R1

S21H

H15 P

akistan 2010 1

2574-CDC China 1980 1

DTU2013 1361 PRJ941 Vibrio cholerae P1 V cholerae El tor Ogawa R1.trim.fq.sorted

F16K

TH6

Pak

ista

n 20

10 1

4585 India 2007 R1

6215

Ken

ya 20

05 R

1

2012EL-1410 Haiti 2012 R1

F17K

TH4

Pak

ista

n 20

10 1

S19

HH

5 P

akis

tan

2010

1

VC51 India 1992 1

F2D

59 P

akis

tan

2010

1

S1K

CH

15 Pakistan 2010 1

S10P

57 Pakistan 2010 1

DTU2016-1930-PRJunkn-Vibrio-cholerae-VC-2 S2 L001 R1 001.trim.fq.sorted

VC-20 Nepal 2010 R1

7686

Ken

ya 2

009

R1

VC-2 Nepal 2010 R1

VC-14 Nepal 2010 R1

2010EL-1796 Haiti 2010 R1

GP140 Malaysi

a 1978 1

2010EL-1961 Haiti 2010

SF20013 Bangladesh 1969

HC-48A1 Haiti 2010 R1

4551 India 2007 R1

V011506-C

DC

China 2001 1

147-CD

C C

hina 2002 1

DT

U20

15 4

89 P

RJ1

014

Vib

rio C

hole

rae

EV

R 2

84 R

1

4656 India 2006 R1

A180 Colombia 1992 1

A488S

xx1xxE B

angladesh 2006 R1

HC-61A1 Haiti 2010 R1

2833-CDC China 1979 1

CIR

S 1

01 B

angl

ades

h 20

02

HC-39A1 Haiti 2010 R1

DTU2013 914 PRJ923 Vibrio cholera 011 17A R1

2010EL-1791 Haiti 2010 R1

DTU2013 920 PRJ923 Vibrio cholera 089 10A R1

6194

Ken

ya 20

07 R

1

A27 Peru 1991 1

2580-CDC China 1989 1

DTU2014 1053 PRJ941 Vibrio cholerae 4053022989 R1.trim.fq.sorted

2011EL-1137 S

outh Africa 2009 R

1

A201 Argentina 1992 1

GP60 In

dia 19

73 1

DTU

2013 1342 PR

J941 Vibrio cholerae V

cholerae El Tor Inaba 22116 R

1.trim.fq.sorted

T21-

CDC Tha

iland

199

0 1

2657-CD

C C

hina 1995 1

2011EL-2322 Haiti 2011 R1

S16H

H1 P

akistan 2010 1

DTU2014 1057 PRJ941 Vibrio cholerae 4053024306 R1.trim.fq.sorted

MB

RN

14 India 2004 R1

HC-23A1 Haiti 2010 R1

VC-15 Nepal 2

010 R1

2011EL-1300 Haiti 2011 R1

DTU2013

133

4 PRJ9

41 V

ibrio

chole

rae

349

264

R1.tri

m.fq

.sorte

d

4663 Bangladesh 2001 R1

VC-3 N

epal 2010 R1

2881-CDC China 1961 1

DTU

2015

-837

-PR

J101

4-Vi

brio

-cho

lera

e-13

ARSC

MC

-018

-706

S2

L001

R1

001.

fast

q.so

rted

DTU

2015

472

PR

J101

4 Vi

brio

Cho

lera

e C

MC

024

R1

2011EL-1089 Haiti 2010

GP143 B

ahrain 197

8 1

200106-CD

C C

hina 2001 1

DTU2013 1363 PRJ941 Vibrio cholerae P17 V cholerae El Tor Ogawa R1.trim.fq.sorted

S5KCH10 Pakistan 2010 1

2757

-CDC C

hina

198

7 1

DTU

2015

497

PR

J101

4 V

ibrio

Cho

lera

e B

RH

121

B R

1

HC-32A1 Haiti 2010 R1

DTU

2015 484 PR

J1014 Vibrio C

holerae EV

R 004 R

12530-C

DC

China 1996 1

4488 India 2006 R1

2921-CDC China 1962 1

F15K

TH7

Pak

ista

n 20

10 1

DTU

2015

500

PR

J101

4 V

ibrio

Cho

lera

e V

SM

658

R1

2009V-1085 USA SxxSri L

ankaxxESxxI

ndiaxxE 2009

VC-25 Nepal 2010 R1

2010EL-1786 Haiti 2010 R1

PC

S-0

23 B

angl

ades

h 20

10 R

1

7687

Ken

ya 2

009

R1

A177 Colombia 1992 1

63244-CDC China 1963 1

DT

U20

15 4

90 P

RJ1

014

Vib

rio C

hole

rae

EV

R 2

85 R

1

HCUF01 Haiti 2010 R1

S4KCH16 Pakistan 2010 1

VC-8 Nepal 2010 R1

DTU

2013

135

6 PR

J941

Vib

rio c

hole

rae

P39

V ch

oler

ae E

l Tor

Inab

a R

1.tri

m.fq

.sor

ted

HC-33A2 Haiti 2010 R1

2010EL-1788 Haiti 2010 R1

6312-CDC Indonesia 1961 12015EL-1160 Kenya unkn 1

S6KCH7 Pakistan 2010 1

DTU2

013

1353

PRJ

941

Vibr

io c

hole

rae

P35

V ch

oler

ae E

l Tor

Inab

a R

1.tri

m.fq

.sor

ted

3500-05 India 2005 R1

A482

Djib

outi

2007

R1

DTU2013 925 PRJ923 Vibrio cholera 196 6A R1

DTU

2013 1344 PR

J941 Vibrio cholerae V

cholerae El Tor Inaba 22118 R

1.trim.fq.sorted

A346 2 Bangladesh 1994 1

2011

EL-

1939

Indi

a 20

09 R

1

SC2000-C

DC

China 2000 1

2010EL-1801 Haiti 2010 R1

HC-38A1 Haiti 2010 R1

1575

-CDC

Chi

na 1

990

1

VC-10 N

epal 2010 R1

DTU

2015

469

PR

J101

4 Vi

brio

Cho

lera

e C

MC

021

R1

MB

N17 India 2004 R

1 F12D

1 P

akis

tan

2010

1

DTU

2015

468

PR

J101

4 Vi

brio

Cho

lera

e C

MC

020

R1

A487Sxx1xxE Bangladesh 2007 R1

CP1042 Thailand 2010 R1

GP145 India 1979 1

3582-05 Pakistan 2005 R

12010EL-2010H Haiti 2010

SC1998 242-1-C

DC

China 1998 1

DTU

2015

473

PR

J101

4 Vi

brio

Cho

lera

e C

MC

025

R1

DTU2

013

1352

PRJ

941

Vibr

io c

hole

rae

P34

V ch

oler

ae E

l Tor

Inab

a R

1.tri

m.fq

.sor

ted

4111 Vietnam

2002 R1

6210

Ken

ya 20

07 R

1

2015EL-1162 Kenya unkn 1

DTU2013

1335

PRJ9

41 V

ibrio

chole

rae 5

10 77

R1.t

rim.fq

.sorte

d

VC-26 Nepal 2010 R1

2848

-CDC

Chi

na 1

988

1

2010EL-1749 Cameroon SxxAfrica W

xxE 2010

S25

R22

Pak

ista

n 20

10 1

F11D

4 P

akis

tan

2010

1

S20H

H14 P

akistan 2010 1

2914-CDC China 1966 1

2015EL-1159 Kenya unkn 1

DT

U2015 481 P

RJ1014 V

ibrio Cholerae E

VR

001 R1

HC-7A1 Haiti 2010

S13P

83 Pakistan 2010 1

DTU

2015

508

PR

J101

4 V

ibrio

Cho

lera

e V

SM

720

R1

3554-08 US

A SxxN

epalxxE 2008 20

09V-

1046

US

A S

xxP

akis

tanx

xE 2

009

2873-CDC China 1961 1

1283-CDC China 1984 1

S2KCH17 Pakistan 2010 1

DTU

2015

480

PR

J101

4 Vi

brio

Cho

lera

e C

MC

032

R1

DTU

2013

135

8 PR

J941

Vib

rio c

hole

rae

P43

V ch

oler

ae E

l Tor

Inab

a R

1.tri

m.fq

.sor

ted

DTU2013 924 PRJ923 Vibrio cholera 151 4A R1

4605 India 2007 R1

VC-11 N

epal 2010 R1

DTU

2015

470

PR

J101

4 Vi

brio

Cho

lera

e C

MC

022

R1

2009V-1131 USA SxxIndiaxxE 2009

2957-CDC China 1964 1

G7555 Bangladesh 1969

DTU2013 926 PRJ923 Vibrio cholera 238 5A R1

HC-71A1 Haiti 2010 R1

2009

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116

US

A S

xxP

akis

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C C

hina 1994 1

S18

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2010

1

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4675

Ban

glad

esh

2001

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2015

505

PR

J101

4 V

ibrio

Cho

lera

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717

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A19 B

angla

desh

1971

1

HC-65A1 Haiti 2010 R1

A130 India 1989 1

DTU

2015

502

PR

J101

4 V

ibrio

Cho

lera

e V

SM

660

R1

DTU2013 927 PRJ923 Vibrio cholera 278 1A R1

6311-CDC Indonesia 1961 1

HC-64A1 Haiti 2010 R1

DTU

2015

507

PR

J101

4 V

ibrio

Cho

lera

e V

SM

719

R1

2011EL-2321 Haiti 2011 R1

2783-CDC China 1964 1

HC-40A1 Haiti 2010 R1

132-CDC China 1978 1

4110 Vietnam

1995 1

DTU

2015

474

PR

J101

4 Vi

brio

Cho

lera

e C

MC

026

R1

A31 Peru 1991 1

MG

1160

25 B

angl

ades

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91 1

DTU

2015

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4-Vi

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lera

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fast

q.so

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hina 1977 1

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2013

135

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rio c

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rae

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339

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4593 India 2007 R1

CP

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1551-CDC China 1979 1

4623 India 2007 R1

2011EL-2319 Haiti 2011 R1

4552 India 2007 R1

S7KCH20 Pakistan 2010 1

DTU

2015

492

PR

J101

4 V

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lera

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RH

117

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1

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2015

501

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659

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RJ1014 V

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hina 2000 1

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hina 1993 1

2012V-1001 USA 2011 R1

DTU2016-1929-PRJunkn-Vibrio-cholerae-VC-1 S1 L001 R1 001.trim.fq.sorted

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2011EL-1941 India 2010 R1

DT

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F19KTH

2 Pakistan 2010 1

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1048

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glad

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angl

ades

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DT

U2015 512 P

RJ1014 V

ibrio Cholerae C

MC

091 R1

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2011EL-2320 Haiti 2011 R1

V01-CDC Chile 1991 1

DTU

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China 1993 1

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bia 20

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a 20

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1

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033 R1

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C C

hina 1997 1

DTU

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506

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J101

4 V

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Cho

lera

e V

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718

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DTU2013 933 PRJ923 Vibrio cholera 330 2A R1

DTU

2013 1341 PR

J941 Vibrio cholerae V

cholerae El Tor Inaba 22115 R

1.trim.fq.sorted

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epal 2010 R1

7682

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hina

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DTU2013 919 PRJ923 Vibrio cholera 068 9A R1

4679

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glad

esh

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akistan 2010 1

VC22

48-1

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C C

hina

200

8 1

2010EL-1792 Haiti 2010

PR

L64 India 1992 1

HC-68A1 Haiti 2010 R1

DTU

2013 1346 PR

J941 Vibrio cholerae V

cholerae El Tor O

gawa 22126 R

1.trim.fq.sorted

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2013

135

4 PR

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rio c

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rae

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oler

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epal 2010 R1

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1338

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01 77

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467

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brio

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lera

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019

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DTU2013 915 PRJ923 Vibrio cholera 021 5A R1

DTU2013 923 PRJ923 Vibrio cholera 150 7A R1

DTU

2015

495

PR

J101

4 V

ibrio

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lera

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2015EL-1158 Kenya unkn 1

A346 1 Bangladesh 1994 1

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C C

hina 2001 1

HC-46A1 Haiti 2010 R1

4672 Bangladesh 2000 R1

3075-CDC China 1974 1

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ista

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DTU2013 1360 PRJ941 Vibrio cholerae P46 V cholerae El Tor Inaba R1.trim.fq.sorted

DTU2013 918 PRJ923 Vibrio cholera 066 13A R1

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1337

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41 V

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chole

rae 8

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rio C

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na 1

991

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TU2013 1345 P

RJ941 V

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l Tor Ogaw

a 22125 R1.trim

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2865-CDC China 1961 1

DTU2013 1362 PRJ941 Vibrio cholerae P8 V cholerae El Tor Ogawa R1.trim.fq.sorted

MJ1236 Bangladesh 1994

SF20011 Bangladesh 1969

VC-22 Nepal 2010 R1

1362 Mozam

bique 2005 R1

DTU2013 917 PRJ923 Vibrio cholera 063 9A R1

2011

EL-

1141

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hani

stan

200

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angl

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akistan 2010 1

2010EL-1799 Haiti 2010 R1

SF20012 Bangladesh 1969

HC-49A2 Haiti 2010 R1

4646 India 2007 R1

HC-70A1 Haiti 2010 R1

2011EL-2317 Haiti 2011 R1

DTU2013 932 PRJ923 Vibrio cholera 315 4A R1

DTU2013 921 PRJ923 Vibrio cholera 094 13A R1

2015EL-1156 Kenya unkn 1

DT

U20

15 4

88 P

RJ1

014

Vib

rio C

hole

rae

EV

R 1

23 R

1

2010EL-2010N Haiti 2010

CP

1050

Ban

glad

esh

2010

R1

CP1041

Zam

bia 20

04 R

1

GP160 India 1980 1

P27457 Bangladesh 1976

4538 India 2007 R1

S9KCH9 Pakistan 2010 1

7684

Ken

ya 2

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2308-CDC China 1978 1

6201

Ken

ya 20

07 R

1

DT

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RJ1014 V

ibrio Cholerae C

MC

089 R1

DTU2013 930 PRJ923 Vibrio cholera 281 8A R1

DTU20

13 1

375

PRJ9

41 V

ibrio

cho

lera

e VC

O1

9 R1.

trim

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orte

d

DTU2013 931 PRJ923 Vibrio cholera 285 11A R1

RND6878 Russia 2012

DT

U20

15 5

14 P

RJ1

014

Vib

rio C

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rae

GM

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31 R

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DTU2

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941

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io c

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rae

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V ch

oler

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l Tor

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DTU2013 1365 PRJ941 Vibrio cholerae P33 V cholerae El Tor Ogawa R1.trim.fq.sorted

3497-CDC China 1980 1

A397 Bangladesh 1987 1

3014-CDC China 1969 1

VC-5 Nepal 2010 R1

DTU

2015

479

PR

J101

4 V

ibrio

Cho

lera

e C

MC

031

R1

2744-CDC China 1962 1

HC-20A2 Haiti 2010

2011EL-2318 Haiti 2011 R1

HC-47A1 Haiti 2010 R1

2981-CDC China 1965 1

DTU

2013

136

4 PR

J941

Vib

rio c

hole

rae

P25

V ch

oler

ae E

l Tor

Oga

wa

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orte

d

DTU

2015

504

PR

J101

4 V

ibrio

Cho

lera

e V

SM

716

R1

A232 Mexico 1991 1

DTU2013 1368 PRJ941 Vibrio cholerae P45 V cholerae El Tor Ogawa R1.trim.fq.sorted

A488S

xx2xxE B

angladesh 2006 R1

HC-43A1 Haiti 2010

DT

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15 4

85 P

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rio C

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rae

EV

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20 R

1

A18 India 1977 1

C6706-CDC Peru 1991 1

A186 Argentina 1992 1

2752-CDC China 1981 1

RND19191 Russia 2001

DTU2013 935 PRJ923 Vibrio cholera 342 3A R1

HC-06A1 Haiti 2010 R1

4210-CD

C C

hina 1999 1

G4222 South Africa 2001

2011EL-2316 Haiti 2011 R1

DTU

2015

493

PR

J101

4 V

ibrio

Cho

lera

e B

RH

118

R1

Country

Philipines

China

Thailand

Pakistan

Bangladesh

India

Nepal

Vietnam

Indonesia

Afghanistan

Bahrain

Malaysia

Haiti

USA

Mexico

Dominican_Republic

Kenya

Djibouti

Mozambique

South_Africa

Zambia

Mauritania

Tanzania

Zimbabwe

Peru

Argentina

Colombia

Bolivia

Chile

Russia

Cameroon

Continent

Asia

North_America

Africa

South_America

Russia

Biotype

El_Tor

El_Tor_variant

Tree scale: 0.01

Philipines

Haiti

Cameroon

Page 17: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Benchmarking of de novo assembly tools: AMR gene detection

Page 18: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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• Take a well characterised set of 383 food pathogen isolates (164 Salmonella and 219 E.coli isolates).

• The set have been assessed for AMR using current gold standard phenotypic tests

• Determine the AMR profile using several of the currently available software for deriving resistance profile from the whole genome sequence data

Objective

Page 19: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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• The genome sequences from the two ENGAGE partner labs were shared in a common data store

• The genome sequences were used to determined genotypic AMR profiles using the following tools:

• ResFinder

• KmerResistance

• SRST2

• PHE genefinder

Methods

Page 20: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Page 21: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Page 22: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Page 23: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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Page 24: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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• The results of the benchmarking showed that predicting antimicrobial resistance using WGS is a feasible and realistic alternative to phenotypic susceptibility testing

• The miscorrelation cases, where the tools predicted a different antimicrobial profile than the expected, were 10 - 14% in Salmonella dataset

• These miscorrelations are suspected to be mistakes in the phenotypic susceptibility test especially the isolates were all tools have identical miscorrelations

Conclusions

Page 25: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

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http://www.engage-europe.eu/resources/benchmarking

Page 26: Tools - ENGAGE benchmarking for AMR gene detection · Tools - ENGAGE benchmarking for AMR gene detection Pimlapas Leekitcharoenphon (Shinny) Research Group of Genomic Epidemiology,

DTU Food, Technical University of Denmark Add Presentation Title in Footer via ”Insert”; ”Header & Footer”

Thank you for your attention

Pimlapas Leekitcharoenphon (Shinny), PhD

Research Group Genomic Epidemiology

WHO Collaborating Centre for Antimicrobial Resistance in Food borne Pathogens

and Genomics

European Union Reference Laboratory for Antimicrobial Resistance

National Food Institute, Technical University of Denmark

[email protected]

26