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Chapter 5.14 : 5.14 T T A B A

Editor: Amy L. Leber1
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Affiliations: 1: Editor in chief, Department of Laboratory Medicine, Nationwide Children’s Hospital, Columbus, Ohio
Content Type: Reference
Publication Year: 2016

Category: Clinical Microbiology

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5.14 T T A B A, Page 1 of 2

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Abstract:

The minimum bactericidal concentration (MBC) test can be used to assess the ability of an antimicrobial agent to kill a bacterial isolate. MBC tests are performed after consultation in very select clinical situations when it is necessary to determine the bactericidal activity of an antimicrobial agent against a bacterial isolate. These situations include those in which immune mechanisms offer little help in eradicating the infecting organisms, such as in patients with endocarditis or osteomyelitis or in immunosuppressed patients, particularly those with neutropenia. Bactericidal tests provide a rough prediction of bacterial eradication, but other factors may also impact the bacteriologic responses of patients. MBC testing is an accepted parameter in the evaluation of new antimicrobial agents and is frequently used as a research tool.

Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14
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5.14 Tests To Assess Bactericidal Activity
Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14
/content/10.1128/9781555818814.chap5.14.fig0019
MIC/MBC QC
MIC/MBC QC
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10.1128/9781555818814/fig0019.gif
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MIC/MBC QC

MIC/MBC QC

Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14
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MBC Worksheet
MBC Worksheet
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MBC Worksheet

MBC Worksheet

Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14
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Figure 5.14.2–A1
Dilution scheme for bacterial colony counts.
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Image of Figure 5.14.2–A1
Figure 5.14.2–A1

Dilution scheme for bacterial colony counts.

Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14
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Figure 5.14.2–A2
Plate configuration for inoculating 0.025 ml of each dilution from reaction tube to agar plate for colony counts. (Adapted and reprinted with permission from Pillai SK, Moellering Jr RC Jr, Eliopoulos GM. 2005. Antimicrobial combinations, p 365–440, Fig. 9.4 [p 373]. Lorian V [ed], , 5th ed. The Williams & Wilkins Co, New York, NY. doi:10.1128/9781555818814.ch5.14.2.fA2
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10.1128/9781555818814/fig5.14.2-a2.gif
Image of Figure 5.14.2–A2
Figure 5.14.2–A2

Plate configuration for inoculating 0.025 ml of each dilution from reaction tube to agar plate for colony counts. (Adapted and reprinted with permission from Pillai SK, Moellering Jr RC Jr, Eliopoulos GM. 2005. Antimicrobial combinations, p 365–440, Fig. 9.4 [p 373]. Lorian V [ed], , 5th ed. The Williams & Wilkins Co, New York, NY. doi:10.1128/9781555818814.ch5.14.2.fA2

Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14
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Time-Kill Assay Worksheet
Time-Kill Assay Worksheet
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Time-Kill Assay Worksheet

Time-Kill Assay Worksheet

Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14
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Time-Kill Assay Worksheet
Time-Kill Assay Worksheet
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Time-Kill Assay Worksheet

Time-Kill Assay Worksheet

Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14
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References

/content/book/10.1128/9781555818814.chap5.14
1. CLSI. 2015. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically, 10th ed. Approved standard M7-A10. CLSI, Wayne, PA.
2. NCCLS. 1999. Methods for Determining Bactericidal Activity of Antimicrobial Agents. Approved guideline. NCCLS document M26-A. NCCLS, Wayne, PA.
3. CLSI. 2015. Performance Standards for Antimicrobial Susceptibility Testing, 25th informational supplement. Approved supplement M100-S25. CLSI, Wayne, PA.
4. Reimer LG, Stratton CW, Reller LB. 1981. Minimum inhibitory and bactericidal concentrations of 44 antimicrobial agents against three standard control strains in broth with and without human serum. Antimicrob Agents Chemother 19: 1050– 1055.
5. Pearson RD, Steigbigel RT, Davis HT, Chapman SW. 1980. Method for reliable determination of minimal lethal antibiotic concentrations. Antimicrob Agents Chemother 18: 699– 708.
6. Taylor PC, Schoenknecht FD, Sherris JC, Linner EC. 1983. Determination of minimal bactericidal concentrations of oxacillin for Staphylococcus aureus: influence and significance of technical factors. Antimicrob Agents Chemother 23: 142– 150.
7. Gunnison JB, Fraher MA, Jawetz E. 1963. Persistence of Staphylococcus aureus in penicillin in vitro. J Gen Microbiol 34: 335– 349.
8. Woolfrey BF, Lally RT, Ederer MN. 1985. Evaluation of oxacillin tolerance in Staphylococcus aureus isolates by a novel method. Antimicrob Agents Chemother 28: 381– 388.
9. Eagle H, Musselman AD. 1948. The rate of bacterial action of penicillin in vitro as a function of its concentration, and its paradoxically reduced activity at high concentrations against certain organisms. J Exp Med 88: 99– 131.
10. Handwerger S, Tomasz A. 1985. Antibiotic tolerance among clinical isolates of bacteria. Rev Infect Dis 7: 368– 386.
11. Shanholtzer CJ, Peterson LR, Mohn ML, Moody JA, Gerding DN. 1984. MBCs for Staphylococcus aureus as determined by macrodilution and microdilution techniques. Antimicrob Agents Chemother 26: 214– 219.
12. Peterson LR, Shanholtzer CJ. 1992. Tests for bactericidal effects of antimicrobial agents: technical performance and clinical relevance. Clin Microbiol Rev 5: 420– 432.
13. Moody J, Knapp C. 2010. Minimum bactericidal concentration testing. In Garcia LS (ed), Clinical Microbiology Procedures Handbook, 3 ed. ASM, Press, Washington, DC.
14. Amsterdam D. 2005. Susceptibility testing of antimicrobials in liquid media, p 61–143. In Lorian V (ed), Antibiotics in Laboratory Medicine, 5th ed. The Williams & Wilkins Co, New York, NY.
15. Gorbach SL, Bartlett JG, Blacklow NB. 2004. Bactericidal testing, p 142. In Gorbach SL (ed). Infectious Diseases, 3rd ed. Lippincott Williams & Wilkins, Philadelphia, PA.
1. CLSI. 2010. Methods for Antimicrobial Dilution and Disk Susceptibility Testing of Infrequently Isolated or Fastidious Bacteria. Approved standard M45-A2. CLSI, Wayne, PA.
1. NCCLS. 1999. Methods for Determining Bactericidal Activity of Antimicrobial Agents. Approved guideline M26-A. NCCLS, Wayne, PA.
2. Woolfrey BF, Lally RT, Ederer MN. 1985. Evaluation of oxacillin tolerance in Staphylococcus aureus isolates by a novel method. Antimicrob Agents Chemother 28: 381– 388.
3. Caron F, Gold HS, Wennerstein CB, Farris MG, Moellering RC, Eliopoulos GM. 1997. Influence of erythromycin resistance, inoculum growth phase and incubation time on assessment of the bactericidal activity of RP 59500 (quinupristin-dalfopristin) against vancomycin-resistant Enterococcus faecium. Antimicrob Agents Chemother 41: 2749– 2753.
4. Moody J, Knapp C. 2010. Time-kill assay. In Garcia LS (ed), Clinical Microbiology Procedures Handbook, 3 ed. ASM Press, Washington, DC.
21. Messick CR, Rodvold KA, Pendland SL. 1999. Modified time-kill assay against multidrug-resistant Enterococcus faecium with novel antimicrobial combinations. J Antimicrob Chemother 44: 831– 834.
22. Zhou Y, Hou Z, Fang C, Xue X, Da F, Wang Y, Bai H, Luo X. 2013. Comparison of microplate and macrodilution methods in time-kill study of new antimicrobial drugs. Folia Microbiol 58: 9– 16.
1. CLSI. 2015. Methods for Dilution and Disk Susceptibility Testing of Infrequently encountered or Fastidious Bacteria. Approved Guideline. 3rd ed. Approved Guideline M45-A3. CLSI, Wayne, PA.
1. Pillai SK, Moellering RC Jr, Eliopoulos GM. 2005. Antimicrobial combinations, p 365–440. In Lorian V (ed], Antibiotics in Laboratory Medicine, 5th ed. The Williams & Wilkins Co, New York, NY.
1. NCCLS. 1999. Methods for Determining Bactericidal Activity of Antimicrobial Agents. Approved guideline M26-A. NCCLS, Wayne, PA.
2. Eliopoulos GM, Eliopoulos CT. 1988. Antibiotic combinations: should they be tested? Clin Microbiol Rev 1: 139– 156.
3. Pillai SK, Moellering RC Jr, Eliopoulos GM. 2005. Antimicrobial combinations, p 365–440. In Lorian V (ed), Antibiotics in Laboratory Medicine, 5th ed. The Williams & Wilkins Co, New York, NY.
4. Moody J, Knapp C. 2010. Time-kill assay for determining synergy. In Garcia LS (ed), Clinical Microbiology Procedures Handbook, 3 ed. ASM Press, Washington, DC.

Tables

5.14 Tests To Assess Bactericidal Activity
Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14
/content/10.1128/9781555818814.chap5.14.table0073
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Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14
/content/10.1128/9781555818814.chap5.14.table0074
Untitled
Generic image for table
Untitled

Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14
/content/10.1128/9781555818814.chap5.14.table0075
MBC testing conditions for various bacteria for microdilution and macrodilution proceduresa
MBC testing conditions for various bacteria for microdilution and macrodilution procedures
Generic image for table
MBC testing conditions for various bacteria for microdilution and macrodilution proceduresa

MBC testing conditions for various bacteria for microdilution and macrodilution procedures

Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14
/content/10.1128/9781555818814.chap5.14.table0076
Rejection value and calculated sensitivity and specificity for each initial concentration based on duplicate 0.01-ml samplesa
Rejection value and calculated sensitivity and specificity for each initial concentration based on duplicate 0.01-ml samples
Generic image for table
Rejection value and calculated sensitivity and specificity for each initial concentration based on duplicate 0.01-ml samplesa

Rejection value and calculated sensitivity and specificity for each initial concentration based on duplicate 0.01-ml samples

Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14
/content/10.1128/9781555818814.chap5.14.table0077
MBC testing conditions for various bacteria for microdilution and macrodilution proceduresa
MBC testing conditions for various bacteria for microdilution and macrodilution procedures
Generic image for table
MBC testing conditions for various bacteria for microdilution and macrodilution proceduresa

MBC testing conditions for various bacteria for microdilution and macrodilution procedures

Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14
/content/10.1128/9781555818814.chap5.14.table0078
Untitled
Generic image for table
Untitled

Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14
/content/10.1128/9781555818814.chap5.14.table0079
Untitled
Generic image for table
Untitled

Citation: Leber A. 2016. 5.14 T T A B A, p 5.14.1.1-5.14.3.6. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch5.14

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