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Cephalosporinase - Plates

Requirements for Plates !

• Inactivation of a broad spectrum of cephalosporins

• Inactivation of residuals of disinfectants (especially contact plates)

ß-Lactam-Antibiotics

• PenicillinsAmpicillin, Mezlocillin, Penicillin

• 1st Generation Cephalosporins: Cefazolin, Cefalotin, Cefalexim

• 2nd Generation Cephalosporins: Cefamandol, Cefuroxim, Cefoxitin (Cefomycin or Methoxa-Cephalosporine)

• 3rd Generation Cephalosporins:Cefotaxim, Ceftriaxon (Rocephin), Ceftazidim

• 4th Generation Cephalosporins:• Cefquinom, Cefepim

• Carbapenems: Meropenem, Imipenem, Ertapenem

Activity of ß-Lactamase I

• ß-Lactamase I (= Penicillinase or Penase) hydrolyses the ß-Lactam ring of Penicillins

• Definition - Activity of Penicillinase: 1 Unit = Cleavage of 1 Micromol

Penicillin G per minute at pH 7 and 25°C

Activity of ß-Lactamase II

• ß-Lactamase II (= Cephalosporinase or Cephase) hydrolyses the ß-lactam ring of Cephalosporins

• Definition - Activity of ß-Lactamase II: 1 Unit = Cleavage of 1 Micromol

Cephalosporin C per minute at pH 7 and 25°C

Activity of different ß-Lactamase II Media - Tests using antibiotic

discs -Inactivation of different Cephalosporins

(test strain S. aureus ATCC 6538 / (antibiotic discs)

10

14

34

1920

43

24 25 25

31

0 0 0 0 0 0

13

10

23

28

0

75

13 12

21

14

18

24

30

-1

4

9

14

19

24

29

34

39

44

49

Cefexim 5 Cefazolin 30 Meropenem 10 Cephotaxim 30 Ceftriaxon 30 Imipenem 10 Cefuroxim 30 Cefquinom 10 Cefepim 30 Cefoxitin 30

diam

eter

of i

nhib

ition

zon

e (m

m)

TSA-Contact +LTHTh+ßLactamaseII - ICR (2281e) TSA Contact +LT +ßLactamaseII - ICR+ (825) Biomerieux 3P Contact TSA+Neutr.+ßLact.

Activity of different ß-Lactamase II Media - Tests using anitbiotic

discs -Inactivation of different Cephalosporins

(test strain S. aureus ATCC 6538 / (antibiotic discs)

0 0 0

14 14

22

16

19

27

41

0 0 0

13 13

1715

19

26

33

0 0 0

1113

23

16

19

27

34

-1

4

9

14

19

24

29

34

39

44

Cefexim 5 Cefazolin 30 Meropenem 10 Cephotaxim 30 Ceftriaxon 30 Imipenem 10 Cefuroxim 30 Cefquinom 10 Cefepim 30 Cefoxitin 30

diam

eter

of i

nhib

ition

zon

e (m

m)

TSA+ß-Lactamase II -ICR (03079e) TSA+LT+ßLactamaseII -ICR (03080e) Biomerieux 3P Sedimentation TSA+Neutr.+ßLact.

Summary of spectrum of efficacy for TSA Contact +LT +ß-Lactamase II

(Article number 825) • Good inactivation in antibiotic disc experiments:

– Cefixim 5 µg /disc– Cefazolin 30 µg /disc – Meropenem 10 µg /disc – Cefotaxim 30 µg /disc – Ceftriaxon 30 µg /disc – Imipenem 10 µg /disc

• Moderate inactivation in antibiotic disc experiments:– Cefuroxim 30 µg /disc – Cefquinom 10 µg /disc

• Poor or missing inactivation in antibiotic disc experiments:– Cefepim 30 µg /disc – Cefoxitin 30 µg /disc

Developement of new Cephalosporinase with a

broad inactivation spectrum

LACTAMATORTM

Inactivating Properties of Different ß-Lactamase II Media -

Tests using Antibiotic Discs and S. aureus -

28

25

28

24

31

46

36

31

27

3234

32

38

25

21

18

21 21

40

29 2927

0 0 0 0

27

12

0 0

11

0 0

11

27

0 0 0

16

0 0 0 0 0 0 0 0 0 0 0 0 0

-1

5

11

17

23

29

35

41

47

Cefep

im (3

0 µg

)

Cefqu

inom

(10

µg)

Cefot

axim

(30 µ

g)

Ceftri

axon

(30

µg)

Cefur

oxim

(30

µg)

Imip

enem

(10

µg)

Mer

open

em (1

0 µg

)

Ertape

nem (1

0 µg

)

Cefox

itin (3

0 µg

)

Penici

llin (1

0 µg)

Ampic

illin

(25

µg)

Mez

lozilli

n (3

0 µg

)

Cefaz

olin

(30

µg)

Inh

ibit

ion

dia

met

er (

mm

)

TSA Contact + LTHTh - ICR+(article 820, withoutCephalosporinase)

TSA Contact + LTHTh + ß-Lactamase II - ICR (article2281e)

TSA Contact + LT + ß-Lactamase II - ICR+ (article825)

TSA Contact + LT +Cephase - ICR+ (article 823)

Method:1. Inoculation of >106 CFU of S. aureus ATCC 6538 - 2. Antibiotic disc is applied - 3. Incubation for 1d at 34°C - 4. Determination of inhibition zone (diameter in mm)

Inactivating Properties of Different ß-Lactamase II Media -

Tests using Antibiotic Discs and K. rhizophila -

38

34

3836

41

44

35

30

35

42

45

40

31

39

31 3032

29

40

26

29

33

0 0 0

12

39

25

1412

19

14

0

12

34

0 0 0

14

0 0 0 0 0 0 0 0

13

0 0 0 0

-1

5

11

17

23

29

35

41

47

Inh

ibit

ion

dia

mete

r (m

m)

TSA Contact + LTHTh - ICR+(article 820, withoutCephalosporinase)

TSA Contact + LTHTh +ß-Lactamase II - ICR (article2281e)

TSA Contact + LT + ß-Lactamase II - ICR+ (article825)

TSA Contact + LT +Cephase - ICR+ (article 823)

Method:1. Inoculation of >106 CFU of K. rhizophila ATCC 9341 - 2. Antibiotic disc is applied - 3. Incubation for 1d at 34°C - 4. Determination of inhibition zone (diameter in mm)

Summary of spectrum of efficacy for TSA Contact +LT +Cephase ICR+

(Article number 823) Good inactivation of the following antibiotics:

– Penicillins: • Ampicillin 25 µg, Mezlocillin 30 µg, Penicillin 10 µg

– Cephalosporins 1st generation: • Cefazolin 30 µg /disc

– Cephalosporins 2nd generation:• Cefuroxim 30 µg, Cefoxitin 30 µg (Cephamycin)

– Cephalosporins 3rd generation:• Cefotaxim 30 µg, Ceftriaxon 30 µg

– Cephalosporins 4th generation:• Cefepim 30 µg, Cefquinom 10 µg

– Carbapenems:• Ertapenem 10 µg, Imipenem, 10 µg, Meropenem 10 µg

New Products

• TSA Contact +LT + Cephase – ICR+ - article number 823Product available for advance orders

shelf life of 4 months completed in June 2009

• TSA +LT + Cephase – ICR – article number 03085eProduct available for advance orders

shelf life of 4 months completed end of July 2009

• LACTAMATORTM LCT-5 • (10 vials each with 50 IU Cephalosporinase and 500 IU

Penicillinase, lyophilised)Product available in April 2009

• LACTAMATORTM LCT-20• (1 vial with 2 kIU Cephalosporinase and 20 kIU

Penicillinase, lyophilised)Product available in April 2009