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Chapter 5.16 : Synergism Testing: Broth Microdilution Checkerboard and Broth Macrodilution Methods

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Synergism Testing: Broth Microdilution Checkerboard and Broth Macrodilution Methods, Page 1 of 2

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Abstract:

Microbiologists commonly use a two-dimensional, two-agent broth microdilution checkerboard to evaluate combinations of antimicrobial agents against microorganisms. A broth macrodilution limited series, agar dilution method, or disk diffusion method can also be used. Test methods are based on CLSI broth dilution susceptibility methods ( ) for evaluating the inhibitory or bactericidal activity ( ) of specific concentrations in combination at a fixed time. interactions are calculated algebraically and interpreted as synergistic, indifferent, or antagonistic depending on whether the antibacterial activity of the combination is greater than, equivalent to, or less than the activities of the individual agents, respectively. interactions can also be represented geometrically with isobolograms ( ).

Citation: Leber A. 2016. Synergism Testing: Broth Microdilution Checkerboard and Broth Macrodilution Methods, p 5.16.1-5.16.23. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.16
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Figures

Image of Example of Broth Macrodilution Showing Synergism
Example of Broth Macrodilution Showing Synergism

Example of Broth Macrodilution Showing Synergism

Citation: Leber A. 2016. Synergism Testing: Broth Microdilution Checkerboard and Broth Macrodilution Methods, p 5.16.1-5.16.23. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.16
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Image of Example of Broth Macrodilution Showing Antagonism
Example of Broth Macrodilution Showing Antagonism

Example of Broth Macrodilution Showing Antagonism

Citation: Leber A. 2016. Synergism Testing: Broth Microdilution Checkerboard and Broth Macrodilution Methods, p 5.16.1-5.16.23. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.16
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References

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1. CLSI. 2012. Methods for Dilution Susceptibility Tests for Bacteria That Grow Aerobically, 9th ed. Approved standard M7-A9. CLSI, Wayne, PA.
2. NCCLS. 1999. Methods for Determining Bactericidal Activity of Antimicrobial Agents; Approved Guideline. NCCLS document M26-A. NCCLS, Wayne, PA.
3. Berenbaum MC. 1978. A method for testing synergy with any number of agents. J Infect Dis 137:122130.
4. Eliopoulos GM, Eliopoulos CT. 1988. Antibiotic combinations: should they be tested? Clin Microbiol Rev 1:139156.
5. Pillai SK, Moellering RCJr, Eliopoulos GM. 2005. Antimicrobial combinations, p 365–440. In Lorian V (ed), Antibiotics in Laboratory Medicine, 5th ed. Lippincott Williams & Wilkins Co., Philadelphia, PA.
6. Rahal JJJr. 1978. Antibiotic combinations: the clinical relevance of synergy and antagonism. Medicine 57:179195.
7. Moody J. 2010. Synergism testing: broth microdilution checkerboard and broth macrodilution methods, p 5.12.1–5.12.21. In Garcia LS (ed), Clinical Microbiology Procedures Handbook, 3 ed. ASM Press, Washington, DC.
8. Weinstein AJ. 1977. Principles of concomitant antibiotic therapy. Med J Aust Spec Suppl 2:1922.
9. Aaron SD, Vandemheen KL, Ferris W, Fergusson D, Tullis E, Haase D, Berthiaume Y, Brown N, Wilcox P, Yozghatlian V, Bye P, Bells S, Chan F, Rose B, Jeanneret A, Stephenson A, Noseworthy M, Freitag A, Paterson N, Doucette S, Harbour C, Ruel M, MacDonald N. 2005. Combination antibiotic susceptibility testing to treat exacerbations of cystic fibrosis associated with multiresistant bacteria: a randomized, double-blind, controlled clinical trial. Lancet 366:463471.
10. Bayer A, Morrison JO. 1984. Disparity between time-kill and checkerboard methods for determination of in vitro bactericidal interaction of vancomycin plus rifampin versus methicillin-susceptible and-resistant Staphylococcus aureus. Antimicrob Agents Chemother 26:220223.
11. Petersen PJ, Labthavikul P, Jones CH, Bradford PA. 2006. In vitro antibacterial activities of tigecycline in combination with other antimicrobial agents determined by chequerboard and time-kill kinetic analysis. J Antimicrob Chemother 57:573576.
12. Rand KH, Houck HJ, Brown P, Bennett D.. 1993. Reproducibility of the microdilution checkerboard method for antibiotic synergy. Antimicrob Agents Chemother 37:613615.
13. Greco WR, Bravo G, Parsons JC. 1995. The search for synergy: a critical review from a response surface perspective. Pharmacol Rev 47: 331385.
14. Johnson MD, MacDougall C, Ostrosky-Zeichner L, Perfect JR, Rex JH. 2004. Combination antifungal therapy. Antimicrob Agents Chemother 48: 693715.
15. Moellering RCJr. 1979. Antimicrobial synergism--an elusive concept. J Infect Dis 140:639641.
16. Odds FC. 2003. Synergy, antagonism, and what the chequerboard puts between them. J Antimicrob Chemother 52:1. doi: 10.1093/jac/dkg301
1. Eliopoulos GM, Moellering RCJr. 1991. Antimicrobial combinations, p 432–492, Fig. 13.2. In Lorian V (ed), Antibiotics in Laboratory Medicine, 3rd ed. The Williams & Wilkins Co., Baltimore, MD.
1. Pillai SK, Moellering RCJr, Eliopoulos GM. 2005. Antimicrobial combinations, p 365–440. In Lorian V (ed), Antibiotics in Laboratory Medicine, 5th ed. Lippincott Williams & Wilkins Co., Philadelphia, PA.

Tables

Generic image for table
Examples of reported combination interactions

Examples of reported combination interactions

Citation: Leber A. 2016. Synergism Testing: Broth Microdilution Checkerboard and Broth Macrodilution Methods, p 5.16.1-5.16.23. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.16
Generic image for table
Example of format of broth microdilution checkerboard panela

Example of format of broth microdilution checkerboard panel

Citation: Leber A. 2016. Synergism Testing: Broth Microdilution Checkerboard and Broth Macrodilution Methods, p 5.16.1-5.16.23. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.16
Generic image for table
Dilution schematics for two-agent broth microdilution checkerboard

Dilution schematics for two-agent broth microdilution checkerboard

Citation: Leber A. 2016. Synergism Testing: Broth Microdilution Checkerboard and Broth Macrodilution Methods, p 5.16.1-5.16.23. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.16
Generic image for table
Untitled

Citation: Leber A. 2016. Synergism Testing: Broth Microdilution Checkerboard and Broth Macrodilution Methods, p 5.16.1-5.16.23. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.16
Generic image for table
Untitled

Citation: Leber A. 2016. Synergism Testing: Broth Microdilution Checkerboard and Broth Macrodilution Methods, p 5.16.1-5.16.23. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.16
Generic image for table
Dilution schematics for two-agent broth macrodilution limited checkerboard

Dilution schematics for two-agent broth macrodilution limited checkerboard

Citation: Leber A. 2016. Synergism Testing: Broth Microdilution Checkerboard and Broth Macrodilution Methods, p 5.16.1-5.16.23. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.16

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