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Page 1: ß-lactamases Clinical Impact and Epidemiology Professor ... · Professor Sebastian Amyes 3 The screen versions of these slides have full details of copyright and acknowledgements

ß-lactamasesClinical Impact and Epidemiology

Professor Sebastian Amyes

1The screen versions of these slides have full details of copyright and acknowledgements

β-lactamasesClinical Impact and Epidemiology

1

Professor Sebastian AmyesUniversity of Edinburgh

EdinburghScotland

β-lactamases in Gram-negative bacteria

>750 β-lactamases reported

…of clinical importance

Cephalosporins

2

Transferable class C β-lactamases Extended-spectrum β-lactamases (ESBLs)

CarbapenemsTransferable class B carbapenamsesTransferable class D carbapenamses

β-lactamase action

1. HO

H NH

N

SN

OO

2

N

S

H

3

Class & active site

Associatedsubstrate

ASerine

BMetal ion

CSerine

DSerine

Penicillin Carbapenem Cephalosporin Penicillin

COO-

β-lactamase

COO-H

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ß-lactamasesClinical Impact and Epidemiology

Professor Sebastian Amyes

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1. InductionInterference with repressor protein

2 De-repression

Enterobacter sppChromosomal β-lactamase expression

Chromosome

4

2. De repressionMutation in R gene

P Rβ-lactamase

Repressor

Emergence of the BIL-1 β-lactamaseMobilisation of chromosomal class C genes onto plasmids

ChromosomePlasmid

ResistantEnterobacteriaceae

ChromosomePlasmid

Klebsiella spp or Escherichia coli

5

P Rβ-lactamase

Dendrogram for AmpC β-lactamasesplasmid and chromsomal

E. coli K12

Ent. cloacaeACT-1

MIR-1

Ent. aerogenes

Citrobacter sppA. baumannii RYC

P. immobilis A5

M. morganii Y. enterocolitica IP97

DHA-2DHA-1 GN7471

1113

BIL-1 CMY-4 CMY-7CMY-3 CMY-6 LAT-4LAT-3 CMY-5 LAT-1

GN346

P99

6Data from: Philippon et al., (2002). Antimicrob. Agents Chemother. 46:1

Aeromonas spp

P. aeurginosa PAO1

O. anthropiP. stuartii VDG96

H. alvei

L. lactamgenus YK90

SLH62ACC-1

SLS73 SST-1 SRT-182116

2171

MOX-1MOX-2

CMY-1CMY-8CMY-9

CepS - A. sobria

CepH - A. hydrophila

AER14 - A. sobria

FOX-3FOX-4FOX-2FOX-5FOX-1

S. marcescens

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ß-lactamasesClinical Impact and Epidemiology

Professor Sebastian Amyes

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Transferable AmpC β-lactamasesCMY ACC ACT FOX MIR MOX Others CMY-1 ACC-1 ACT-1 FOX-1 MIR-1 MOX-1 CFE-1 CMY-2 ACC-2 ACT-2 FOX-2 MIR-2 MOX-2 CMY-3 ACC-3 ACT-3 FOX-3 MIR-3 MOX-3 DHA-1 CMY-4 ACC-4 ACT-4 FOX-4 MIR-4 MOX-4 DHA-2 CMY-5 ACT-5 FOX-5 MIR-5 MOX-5 CMY-6 ACT-6 FOX-6 MOX-6 LAT-1CMY-7 ACT-7 FOX-7 MOX-7CMY-8 CMY-9 CMY-10 CMY-11 CMY-12 CMY 13

7Total ~ 78

CMY-13 CMY-14 CMY-15 CMY-16 CMY-17 CMY-18 CMY-19 CMY-20 CMY-21 CMY-22

¦¦¦

CMY-44

~44 4 ~7 7 5 7 4

What are extended-spectrum β-lactamases?

“Extended-Spectrum β-lactamases (ESBLs) are enzymes that can be produced by bacteria making them resistant to cephalosporins”

8

making them resistant to cephalosporins

Health Protection Agency

http://www.hpa.org.uk/infections/topics_az/esbl/default.htm

OXA 1

TEM-2

SHV-1

Dominance of the TEM-1 β-lactamase

Accounts for 75% of all plasmid-encoded enzymes

9

OXA-1

PSE-1CARB-6

TEM-1OXA-2

SAR-2

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ß-lactamasesClinical Impact and Epidemiology

Professor Sebastian Amyes

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What happened in Liverpool in 1982?

TEM-1

Extensive ceftazidime use

10

TEM-1 TEM-1

Gene duplication –small increase in MIC

TEM-1 TEM-12

Mutation at codon 164 –MIC increases to 4mg/L

ß-3ß-4

α-10α-2

α-9

Binding of amoxycillin

Active site of the TEM-1 β-lactamase

11

α-7

α-3

α-5

α-4

Ω-loop

Ser-70

Asp-179

Arg-164

Glu-171

ß-3ß-4

α-10α-2

α-9

Active site of the TEM-1 β-lactamaseFailure to bind cephalosporins

12

α-7

α-3

α-5

α-4 Ser-70

Arg-164

Asp-179

Glu-171 Ω-loop

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ß-lactamasesClinical Impact and Epidemiology

Professor Sebastian Amyes

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ß-3ß-4

α-10α -2

α-9

Serine for arginine substitution at 164 (TEM-12)

13

α-7

α-3

α-5

α-4 Ser-70

Ser-164

Asp-179

Glu-171

Amino acid substitutions in TEM ESBLs

Leu21

Gln39

Ala42

Leu51

Gly92

Glu104

His153

Arg164

Met182

Gly218

Ala237

Gly238

Glu240

Arg244

Thr265

Ser268

PheTEM-4TEM-9TEM-25TEM-48TEM-49TEM-53TEM-63

ValTEM-42

AspTEM-57TEM-66

ArgTEM-21TEM-56

ThrTEM-20TEM-43TEM-52TEM-63TEM-72

LysTEM-5TEM-10TEM-24TEM-27TEM-28 TEM-42TEM-46 TEM-47TEM-48

TEM-49TEM-61TEM-68TEM-72

SerTEM-50Thr

TEM-5TEM-24

GlyTEM-22

MetTEM-4TEM-9 TEM-13 TEM-25 TEM-27 TEM-42 TEM-47 TEM-48 TEM-49 TEM-68

TEM-1

14

LysTEM-3TEM-4TEM-6TEM-8TEM-9TEM-14TEM-15TEM-16TEM-17TEM-18TEM-21

TEM-22TEM-24TEM-26TEM-43TEM-46TEM-50TEM-52TEM-56TEM-60TEM-63TEM-66

LysTEM-2TEM-3TEM-7TEM-8TEM-11TEM-13TEM-16TEM-18TEM-21TEM-22TEM-42TEM-46TEM-56TEM-60TEM-61TEM-66TEM-72

ProTEM-60

GluTEM-55

SerTEM-5TEM-7TEM-8TEM-9TEM-10TEM-12TEM-22TEM-25TEM-26TEM-46TEM-53TEM-60 TEM-63

HisTEM-6TEM-11TEM-16 TEM-27 TEM-28 TEM-29 TEM-43 TEM-61

SerTEM-3TEM-4TEM-8TEM-14TEM-15TEM-18TEM-19TEM-20TEM-21TEM-22TEM-25TEM-42TEM-47TEM-48TEM-49TEM-52TEM-66TEM-68TEM-72

LeuTEM-54

SerTEM-58

GlyTEM-49

Data from: Bradford (2001). Clin. Microbiol. Rev. 14:933

Sequential mutation of ESBLsby cephalosporins

3rd generation cephalosporins

R164S E240K A237T

15

TEM-12 TEM-10 TEM-5TEM-1

MIC (mg/l)

Ceftazidime <0.1 4 64 64

Cefotaxime <0.1 <0.1 0.5 16

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ß-lactamasesClinical Impact and Epidemiology

Professor Sebastian Amyes

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Porin

Outside cell

Outer membrane

Interaction of cephalosporins with β-lactamases in Gram-negative bacteria

Ceftazidime

Porin

Ceftriaxone or Cefotaxime

16

Penicillin binding proteinsβ-lactamase

Periplasmicspace

Cytoplasm β-lactamase Penicillin binding proteins

Progression of TEM-1 to TEM-5 β-lactamaseEffect on hydrolysis rate and inhibitor sensitivity

0.01

0.1

1

10

100

s)

CeftazidimeCefotaximeAmpicillinClavulanic acid

17Data from Du Bois, Marriott & Amyes (1995). J. Antimicrob. Chemother. 35: 7

TEM-1 TEM-12 TEM-10 TEM-51

10

100

10000.001

1000

100

10

1

Kca

t(1

/s

Cla

vula

nic

acid

I 50(n

M)

ß-3ß-4

α-10α-2

α-9

Substitutions of the TEM-1 β-lactamase(ESBL formation)

18α-7

α-3

α-5

α-4

Ω-loop

Ser-70

Arg-164

Ala-237

Gly-238

Glu-240Glu-104

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ß-lactamasesClinical Impact and Epidemiology

Professor Sebastian Amyes

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Lys SerLys Pro GluSer His Leu Gly

Amino acid substitutions in TEM ESBLs

Leu21

Gln39

Ala42

Leu51

Gly92

Glu104

His153

Arg164

Met182

Gly218

Ala237

Gly238

Glu240

Arg244

Thr265

Ser268

PheTEM-4TEM-9TEM-25TEM-48TEM-49TEM-53TEM-63

ValTEM-42

AspTEM-57TEM-66

ArgTEM-21TEM-56

ThrTEM-20TEM-43TEM-52TEM-63TEM-72

LysTEM-5TEM-10TEM-24TEM-27TEM-28 TEM-42TEM-46 TEM-47TEM-48

TEM-49TEM-61TEM-68TEM-72

SerTEM-50Thr

TEM-5TEM-24

GlyTEM-22

MetTEM-4TEM-9 TEM-13 TEM-25 TEM-27 TEM-42 TEM-47 TEM-48 TEM-49 TEM-68

TEM-1

19

LysTEM-3TEM-4TEM-6TEM-8TEM-9TEM-14TEM-15TEM-16TEM-17TEM-18TEM-21

TEM-22TEM-24TEM-26TEM-43TEM-46TEM-50TEM-52TEM-56TEM-60TEM-63TEM-66

SerTEM-3TEM-4TEM-8TEM-14TEM-15TEM-18TEM-19TEM-20TEM-21TEM-22TEM-25TEM-42TEM-47TEM-48TEM-49TEM-52TEM-66TEM-68TEM-72

LysTEM-2TEM-3TEM-7TEM-8TEM-11TEM-13TEM-16TEM-18TEM-21TEM-22TEM-42TEM-46TEM-56TEM-60TEM-61TEM-66TEM-72

ProTEM-60

GluTEM-55

SerTEM-5TEM-7TEM-8TEM-9TEM-10TEM-12TEM-22TEM-25TEM-26TEM-46TEM-53TEM-60 TEM-63

HisTEM-6TEM-11TEM-16 TEM-27 TEM-28 TEM-29 TEM-43 TEM-61

LeuTEM-54

SerTEM-58

GlyTEM-49

Data from: Bradford (2001). Clin. Microbiol. Rev. 14:933

ß-3ß-4

α-10α-2

α-9

Substitution of the SHV-1 β-lactamase

Leu-205

20

α-7

α-3

α-5

α-4 Ser-70

Arg-164

Gly-238Glu-240

Interaction of the extended-spectrumSHV β-lactamases

SHV-2

SHV-1

G238S

SHV ?

21

SHV-4

SHV-5

R205L

SHV-3

R205L

E240K

E240K

SHV-?

SHV-?

SHV-7

I8F

I8FR43S

R43S

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ß-lactamasesClinical Impact and Epidemiology

Professor Sebastian Amyes

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Group A

Dendrogram of PFGE patterns in Klebsiella pneumoniae

22

Group BGroup C

Group D

Group E

ESBLs in Klebsiella pneumoniaeRoyal Infirmary Edinburgh 1999 - 2000

Group A 25 SHV-2 (21)SHV-2 + SHV-5 (4)

TEM-1 70 + 141

No ESBLs Other blas Plasmids (kb)

23

Group BGroup C

Group D

Group E

12

4

10

5

SHV-2

SHV-2

SHV-2

SHV-2

-

TEM-1

-

TEM-1

141

70 + 141

141

70 + 141

Extended-spectrum β-lactamasesTEM SHV CTX OXA

TEM-4 SHV-2 CTX-M-1 OXA-10 TEM-5 SHV-2a CTX-M-2 OXA-11 TEM-6 SHV-3 CTX-M-3 OXA-13 TEM-9 SHV-4 CTX-M-4 OXA-14 TEM-10 SHV-5 CTX-M-5 OXA-15 TEM-12 SHV-6 CTX-M-6 OXA-16 TEM-14 SHV-7 CTX-M-7 OXA-17 TEM-15 SHV-8 CTX-M-8 OXA-19 TEM-17 SHV-9 CTX-M-9 OXA-28 TEM-19 SHV-12 CTX-M-10 OXA-32 TEM-20 SHV-13 Toho-1 OXA-35 TEM-24 SHV-14 ¦ TEM-25 SHV-18 ¦ TEM-26 SHV-19 ¦ TEM-27 SHV-20 CTX-M-89 TEM-28 SHV-21 TEM 29 SHV 22

Original enzyme

24

TEM-29 SHV-22 TEM-43 SHV-24 TEM-47 ¦ TEM-48 ¦ TEM-49 ¦ TEM-50 SHV-127

¦ ¦ ¦

TEM-174

~160 ~127 ~89 11

Total ~ 360SlowFast FastSlowType of cephalosporin

Ceftazidime CeftazidimeCefotaxime/Ceftriaxone

Cefotaxime/Ceftriaxone

Example

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ß-lactamasesClinical Impact and Epidemiology

Professor Sebastian Amyes

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Emergence of CTX-M β-lactamases in Ramón y Cajal Hospital, Madrid

30

40

50

CTX-M-32CTX-M-15CTX-M-1/3of

Clo

nes 1st CTX-M-14, CTX-M-1/3

CTX-M-15

1st CTX-M-32

25

0

10

20

1988

198919

90199

1199

21993

1994199

51996

1997

199819

9920

00200

1200

2

CTX-M-14CTX-M-9CTX-M-10SHVTEM

Data from: Canton et al., (2006); Current Op Microbiol. 9: 466

Year

Num

ber

1st CTX-M-91st CTX-M(CTX-M-10)

European spread of CTX-M β-lactamases

CTX-M-2

CTX-M-1, -3, -32

CTX M 9 14 18 19 20 21

CTX-M-1, -3, -32CTX-M-9, -14

CTX-M-1, -3, -10, -15

CTX-M-40

26CTX-M-1, -3, -32

CTX-M-4

CTX-M-15

CTX-M-9, -14, -18, -19, -20, -21

CTX-M-1, -3, -15

CTX-M-2,5

EndemicSporadicNo dataCTX-M-1CTX-M-2CTX-M-8CTX-M-9

Data from: Canton et al., (2006); Current Op Microbiol. 9: 466

CTX-M-1, -10, -15, -32

CTX-M-2

CTX-M-9, -14

Origins of CTX-M β-lactamases

CTX-M-31

CTX-M-6

CTX-M-5Toho-1

CTX-M-20

CTX-M-7CTX-M-4

CTX-M-2 CTX-M-2

CTX-M-27CTX-M-24

CTX M 19

CTX-M-16CTX-M-9

CTX-M-14CTX-M-18

CTX-M-9

27

Toho-2

CTX-M-3

CTX-M-28CTX-M-22

CTX-M-10CTX-M-11

CTX-M-12CTX-M-29

CTX-M-30CTX-M-15CTX-M-33

CTX-M-23CTX-M-1

CTX-M-1

CTX-M-19

CTX-M-13CTX-M-17

CTX-M-21

CTX-M-26CTX-M-25 CTX-M-25

CTX-M-8

CTX-M-8CTX-M-1 CTX-M-2 CTX-M-9 CTX-M-25 CTX-M-8

Year 1989 1986 1996 1994 2000

Country Germany Japan Brazil Spain Canada

Origin K. ascobata K. ascobata K. georgiana K. georgiana

www.lahey.org

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ß-lactamasesClinical Impact and Epidemiology

Professor Sebastian Amyes

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ESBL activity

Vitek 2 tests for extended-spectrum β-lactamases

28

1684 3224126321.51.0.75.5 TZ4 1 .

5

.758 6 3 1 .5 .38

.25

.19

.125

TZ

L testESBL

ε-test for extended-spectrum β-lactamases

Why should we measure ESBL activity?

Many ESBLs are a combination of sequential mutations

Early detection of the initial mutations is essential

S d thi d t ti i id i i MIC

29

Second or third mutations may give very rapid rises in MIC

Ceftazidime and cefotaxime breakpoints are set for clinical efficacy and are too high to detect early ESBL mutations

ESBL-producing Klebsiella pneumoniaeAntibiotic time kill

56789

10

ount

(CFU

/ml) Growth controlPiperacillin-tazobactam

Cefepime

30Data from Burgess & Hall (2004); Diagnostic Microbiology and Infectious Disease 49: 41–46

012345

0 4 8 12 16 20 24

Log

Col

ony

co

Time (hours)

Meropenem

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ß-lactamasesClinical Impact and Epidemiology

Professor Sebastian Amyes

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Carbapenem resistance emerging in a CTX-M containing K. pneumoniae

Outbreak of ESBL producing Klebsiella pneumoniaein a hospital in Mallorca, Spain in 200551 patients infected with single clone –determined by PFGE

31

ESBL identified as CTX-M-1All but 1 strain resistant to ciprofloxacin, gentamicin, co-trimoxazole and tetracyclineIn 2 patients strains resistant to imipenem, meropenem and ertapenemTwo outer membrane proteins lost, Omp35 and Omp36Treatment unspecified!

Data from: Mena et al., (2006); J Clin Microbiol. 44: 2831

The original transferable carbapenemases

Sme-1

Sme-2

NMC-A

IMI-1

Class AS. marcescens

E. cloacae

UK, USA

France, USA

32

IMP

VIM

Class B

Data from Livermore et al., (2000) Curr. Opin. Microbiol 3: 489

24

23

OXA-23 β-lactamase

MIC = 16mg/L

Imipenem resistance in Acinetobacter baumannii from Scotland

33Paton et al., (1993); Int. J. Antimicrob. Agents. 2: 81Scaife et al., (1995); J. Antimicrob. Chemother. 36:585

MIC 16mg/Lβ-lactamase not induced− pI of β-lactamase is 6.65

Gene located on 40kB plasmid & is transferable to A. juniiDNA sequence shows close similarity with OXA β-lactamasesIst plasmid-mediated class D carbapenemaseStrain is sensitive to sulbactam

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ß-lactamasesClinical Impact and Epidemiology

Professor Sebastian Amyes

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Pairwise (OG:100%,UG:0%) (FAST:2,10) Gapcost:3%

100

9590858075706560 EuropeS. AmericaAsia

Dendrogram of OXA β-lactamases in Acinetobacter baumannii

OXA-23

34Brown et al., (2006); J. Antimicrob. Chemother. 57:1-3Rasmussen et al., (2006); J. Antimicrob. Chemother. 57:373-383

EuropeAsiaAfricaS. AmericaN. America

EuropeAsiaS America

OXA-51

EuropeOXA-40

OXA-58

1 OXA-23 MN--KYFTCYVVASLFLSGCTVQHNLIN-ETPSQIVQG--HNQVIHQYFDEKNTSGVLVI 552 OXA-40 MKKFILPIFSISILVSLSACSSIKTKSEDNFHIS-SQQ--HEKAIKSYFDEAQTQGVIII 573 OXA-51 MN--IKTLLLITSAIFISACSPYIVTANPNHSASKSDE--KAEKIKNLFNEVHTTGVLVI 564 OXA-58 MK-LLKILSLVCLSISIGACAEHSMSRAKTSTIPQVNNSIIDQNVQALFNEISADAVFVT 59

1 OXA-23 QTDKKINLYGNALSRANTEYVPASTFKMLNALIGLENQKTDINEIFKWKGEKRSFTAWEK 1152 OXA-40 KEGKNLSTYGNALARANKEYVPASTFKMLNALIGLENHKATTNEIFKWDGKKRTYPMWEK 1173 OXA-51 QQGQTQQSYGNDLARASTEYVPASTFKMLNALIGLEHHKATTTEVFKWDGQKRLFPEWEK 1164 OXA-58 YDGQNIKKYGTHLDRAKTAYIPASTFKIANALIGLENHKATSTEIFKWDGKPRFFKAWDK 119

OXA β-lactamase subgroupsSequence similarities

Subgroup

35

1 OXA-23 DMTLGEAMKLSAVPVYQELARRIGLDLMQKEVKRIGFGNAEIGQQVDNFWLVGPLKVTPI 1752 OXA-40 DMTLGEAMALSAVPVYQELARRTGLELMQKEVKRVNFGNTNIGTQVDNFWLVGPLKITPV 1773 OXA-51 DMTLGDAMKASAIPVYQDLARRIGLELMSKEVKRVGYGNADIGTQVDNFWLVGPLKITPQ 1764 OXA-58 DFTLGEAMQASTVPVYQELARRIGPSLMQSELQRIGYGNMQIGTEVDQFWLKGPLTITPI 179

1 OXA-23 QEVEFVSQLAHTQLPFSEKVQANVKNMLLLEESNGYKIFGKTGWAMDIKPQVGWLTGWVE 2352 OXA-40 QEVNFADDLAHNRLPFKLETQEEVKKMLLIKEVNGSKIYAKSGWGMGVTPQVGWLTGWVE 2373 OXA-51 QEAQFAYKLANKTLPFSPKVQDEVQSMLFIEEKNGNKIYAKSGWGWDVDPQVGWLTGWVV 2364 OXA-58 QEVKFVYDLAQGQLPFKPEVQQQVKEMLYVERRGENRLYAKSGWGMAVDPQVGWYVGFVE 239

1 OXA-23 QPDGKIVAFALNMEMRSEMPASIRNELLMKSLKQLNII--- 2732 OXA-40 QANGKKIPFSLNLEMKEGMSGSIRNEITYKSLENLGII--- 2753 OXA-51 QPQGNIVAFSLNLEMKKGIPSSVRKEITYKSLEQLGIL--- 2744 OXA-58 KADGQVVAFALNMQMKAGDDIALRKQLSLDVLDKLGVFHYL 280

40

50

60

ffici

ency

Carbapenemase efficiency (kcat/Km)related to benzylpenicillin

360

10

20

30

OXA-23 OXA-40 OXA-51 OXA-58Subgroup 1 2 3 4

Perc

enta

ge e

f

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ß-lactamasesClinical Impact and Epidemiology

Professor Sebastian Amyes

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Carbapenem-hydrolysing β-lactamasesClass B metallo β-lactamase Class A serine β-lactamase Class D OXA serine β-lactamase

IMP VIM IND GES KPC SME OXA-23-

like OXA-40-

like OXA-51-

like OXA-58-

like IMP-1 VIM-1 IND-1 GES-1 KPC-1 SME-1 OXA-23 OXA-40 OXA-51 OXA-58IMP-2 VIM-2 IND-2 GES-2 KPC-2 SME-2 OXA-27 (OXA-24) OXA-64 OXA-96IMP-3 VIM-3 IND-2a GES-3 KPC-3 SME-3 OXA-49 OXA-25 OXA-65 OXA-97IMP-4 VIM-4 IND-3 GES-4 KPC-4 OXA-102 OXA-26 OXA-66IMP-5 VIM-5 IND-4 GES-5 KPC-5 OXA-103 OXA-72 OXA-67IMP-6 VIM-6 IND-5 GES-6 KPC-6 OXA-105 OXA-68IMP-7 VIM-7 IND-6 GES-7) KPC-7 OXA-133 OXA-69IMP-8 VIM-8 IND-7 GES-8 KPC-8 OXA-134 OXA-70IMP-9 VIM-9 GES-9 KPC-9 OXA-71IMP-10 VIM-10 GES-10 KPC-10 OXA-73IMP-11 VIM-11 GES-11 OXA-74

37

IMP-11 VIM-11 GES-11 OXA-74IMP-12 VIM-12 GES-12 OXA-75 IMP-13 VIM-13 OXA-76IMP-14 VIM-14 OXA-77IMP-15 VIM-15 OXA-78IMP-16 VIM-16 OXA-79IMP-17 VIM-17 OXA-80IMP-18 VIM-18 OXA-82IMP-19 VIM-19 ¦ IMP-20 VIM-20 ¦ IMP-21 VIM-21 ¦ IMP-22 VIM-22 ¦ IMP-23 VIM-23 ¦ IMP-24 OXA-132 24 23 8 12 10 3 8 4 56 3

55 25 ~ 71

Total > 150

OXA 71

Q194POXA-64

OXA-99

A5TA48V

Q57RK209M

OXA-87 OXA-86 OXA-67A5T

Q194L

V196A T74I

A31TV101I

A12T

T24SOXA-111 K109M

K146N D117N

A12T

OXA-93

G90S

OXA-75

P256SH29Y

K4Q

S267GK266R V151I

OXA-89

P20HA5T

A182T

OXA-78I255TE100AT24S

K

N225D

K

The OXA-51-like β-lactamases

OXA-98

OXA 113 L167V

OXA-51

38

K107EQ57H

OXA-71K107Q

OXA-91 OXA-68

195E

OXA-77

K146N

E36KD105NOXA-106OXA-70

D198H

OXA-94

OXA-80

OXA-83

OXA-84

E36V

Q239H K4Q

P130S

E36K

OXA-109OXA-82

P130Q

OXA-65 OXA-88

OXA-92OXA-69E36D W234S

L167V

OXA-110 OXA-107 OXA-112

A25TOXA-108 A156T

OXA-95

OXA-104 K146ND117N

OXA-79

OXA-76Q270K

OXA-113

OXA-80 P130L

Data from: Evans, Hamouda, Towner & Amyes (2008); Clin Microbiol Infect 14: 268–275

OXA-66

Dice (Opt:1.30%) (Tol 1.3%-1.3%) (H>0.0% S>0.0%) [0.0%-100.0%]Apa1

100

806040

Apa1

A1

A384

A388

A230

A387

A25

A4

Nottingham, United Kingdom

Oslo, Norway

Ioannina, Greece

Nottingham, United Kingdom

Ioannina, Greece

Nottingham, United Kingdom

Nottingham, United Kingdom

0.25

4

1

0.25

4

0.25

0.5

0.5

2

2

1

4

0.5

0.5

8

128

>256

>256

>256

32

32

OXA-69

OXA-69

OXA-92

OXA-66

OXA-66

OXA-66

OXA-66

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

+

+

-

+

-

-

-

+

+

-

+

+

+

-

-

+

-

+

-

-

-

+

+

-

+

-

-

Linkage of specific blaOXA-51-like genes to common multi-resistant clones

39

A6

A24

A13

A20

A478

A37

A397

A47

A320

A297

A392

A60

A377

Nottingham, United Kingdom

Nottingham, United Kingdom

Rotterdam, The Netherlands

Paris, France

LUH 5875 European clone III

Singapore

Thessaloniki, Greece

Berlin, Germany

RUH 134 European clone II

RUH 875 European clone I

Freiburg, Germany

Buenos Aires, Argentina

Regensburg, Germany

0.25

0.12

1

0.25

0.25

0.25

4

0.5

0.12

0.25

8

0.25

4

0.5

0.25

2

1

2

0.25

4

1

0.5

0.5

8

1

2

32

16

>256

32

64

128

>256

16

8

8

32

32

64

OXA-66

OXA-66

OXA-71

OXA-71

OXA-71

OXA-64

OXA-66

OXA-66

OXA-66

OXA-69

OXA-66

OXA-65

OXA-71

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

-

+

-

-

-

+

-

+

+

+

-

-

-

+

-

+

+

+

+

+

-

-

-

-

-

-

-

+

-

-

-

+

-

+

-

-

-

-

-

-

+

-

-

-

+

-

+

Data from: Evans, Hamouda, Towner & Amyes.(2007) ICAAC, Chicago

Page 14: ß-lactamases Clinical Impact and Epidemiology Professor ... · Professor Sebastian Amyes 3 The screen versions of these slides have full details of copyright and acknowledgements

ß-lactamasesClinical Impact and Epidemiology

Professor Sebastian Amyes

14The screen versions of these slides have full details of copyright and acknowledgements

OXA-66

ISAba1

Insertion of ISAba1 OXA-51-like β-lactamases now confer carbapenem resistance

40

OXA-69OXA-71blaOXA-51-like

Promoter

7 bases from start codon

Summary of OXA β-lactamases in Acinetobacter baumannii

Class D carbapenemases are now the major mechanism of carbapenem resistance in Acinetobacter baumanniiOXA-23 now found in 2 clones in 50 hospitals in and around London, EnglandOXA-58 found to be the predominant carbapenemase in parts of Asia

41

OXA-51-like is the largest group of OXA-β-lactamases in this genus; Can causes carbapenem resistance if ISAba1 is presentThe carriage of OXA-51-like β-lactamases in all Acinetobacter baumanniistrains worldwide has serious implications for the treatment of A. baumannii infections Pan-drug resistant Acinetobacter baumannii are prevalent in some hospitals

Acknowlegements

University of DundeeDr Hilary-Kay YoungDr Wendy ScaifeHelen Donald

University of EdinburghDr Susan BrownDr Sarah DaviesBen EvansDr Ahmed HamoudaDr Paul Higgins South Africa

ArgentinaDr Carlos Bantar

EnglandDr Neil WoodfordHPA Colindale

42

ggDr Stephen Du BoisDr Ali DashtiDr Rachel WalkerDr Fiona WalshDr Alexandra RolfeDr John HoodDr Robert PatonDr Rex MilesDr Ian Laurenson

South AfricaProf Heather Crewe-BrowneUSADr Fred TenoverCDC, Atlanta

Page 15: ß-lactamases Clinical Impact and Epidemiology Professor ... · Professor Sebastian Amyes 3 The screen versions of these slides have full details of copyright and acknowledgements

ß-lactamasesClinical Impact and Epidemiology

Professor Sebastian Amyes

15The screen versions of these slides have full details of copyright and acknowledgements

43