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Singapore Society of Pathology Singapore Society of Pathology Society of Infectious Diseases, Singapore Society of Infectious Diseases, Singapore & & Antimicrobial Antimicrobial Susceptibility Susceptibility Testing Testing Faculty members Dr. Koh Tse Hsien Dr. Nancy Tee Mr. Lim Tze Peng Dr. Jaime Chien Dr. Tan Thean Yen Ms. Lily Ng Siew Yong Ms. Tan Peck Lay Ms. Irene Tan organised in conjunction with Department of Pathology, Singapore General Hospital, 21-23 rd September 2010, Singapore Sponsors: We gratefully acknowledge the support of the following organisations: Slides Pictures and text provided courtesy of: Dr. Koh Tse Hsien Singapore General Hospital We also acknowledge support from the following companies: Siemens Healthcare Diagnostics Pte Ltd

Demonstration AST Plates

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Page 1: Demonstration AST Plates

Singapore Society of PathologySingapore Society of Pathology

Society of Infectious Diseases, SingaporeSociety of Infectious Diseases, Singapore&&

AntimicrobialAntimicrobial

SusceptibilitySusceptibility

TestingTesting

Faculty members

Dr. Koh Tse Hsien

Dr. Nancy Tee

Mr. Lim Tze Peng

Dr. Jaime Chien

Dr. Tan Thean Yen

Ms. Lily Ng Siew Yong

Ms. Tan Peck Lay

Ms. Irene Tan

organised in conjunction with

Department of Pathology,

Singapore General Hospital,

21-23rd September 2010,

Singapore

Sponsors:

We gratefully acknowledge the

support of the following

organisations:

Slides

Pictures and text provided

courtesy of:

Dr. Koh Tse Hsien

Singapore General Hospital

We also acknowledge support from the

following companies:

Siemens Healthcare Diagnostics Pte Ltd

Page 2: Demonstration AST Plates

Klebsiella producing DMA plasmid AmpC beta-lactamase. IPI: Imipenem+MBL (metallo-beta-lactamase) inhibitor (EDTA), IP: ImipenemThere is some non-specific decrease in imipenem MIC in the presence of the inhibitor but not as dramatic as what you will see with a true MBL producer. DHA is a class C enzyme that is inhibited by boronic acid and cloxacillin.

Page 3: Demonstration AST Plates

Klebsiella pneumoniae KPC ATCC control strain. There is no significant fall of imipenem in the presence of MBL inhibitor. This is becase KPC is a class A enzyme that is inhibited by clavulanic acid

Page 4: Demonstration AST Plates

Klebsiella class B metallo-beta-lactamase (NDM-1) producer. There is a dramatic decrease in imipenem MIC in thepresence of inhibitor. The inhibitor (EDTA) is a metal chelator that prevents zinc ions from reaching the active site of the enzyme.

Page 5: Demonstration AST Plates

Clockwise from bottom right. Meropenem; Meropenem + Boronic acid; Meropenem +Cloxacillin; Meropenem +EDTAKlebsiella plasmid AmpC DHA producer. Class C enzymes are inhibited by boronic acid and cloxacillin. This strain also shows some non-specific inhibition with EDTA.

Page 6: Demonstration AST Plates

Clockwise from bottom right. Meropenem; Meropenem + EDTA; Meropenem +Cloxacillin; Meropenem +boronic acidKlebsiella plasmid ATCC KPC control. KPC enzymes are only inhibited by boronic acid

Page 7: Demonstration AST Plates

Clockwise from bottom right. Meropenem; Meropenem + boronic acid ; Meropenem +Cloxacillin; Meropenem +EDTA. Klebsiella metallo-beta-lactamase NDM-1 producer. There is a large zone of inhibition around the meropenem disk with added EDTA indicating the presence of a MBL. There is also a suggestion of inhibition around the meropenem disk with added boronic acid. This strain produced both NDM-1 MBL and CMY and DHA plasmid AmpC enzymes-but the MBL is swamping the effects of the plasmid AmpC so that the AmpC phenotype is not seen.


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