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Chapter 68 : Susceptibility Test Methods: Dilution and Disk Diffusion Methods

Authors: Jean B. Patel, Fred C. Tenover, John D. Turnidge, James H. Jorgensen
Content Type: Reference
Publication Year: 2011

Category: Clinical Microbiology

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Abstract:

There are a number of methods for antimicrobial susceptibility testing of bacteria, and they are categorized into dilution methods that generate Minimum Inhibitory Concentration (MIC) results and disk diffusion methods that generate zone diameter results. The Clinical and Laboratory Standards Institute (CLSI) reference methods are broth macrodilution, broth microdilution, agar dilution, and disk diffusion. Dilution methods are also readily adaptable to automated test systems. Stock solutions are prepared as discussed in the CLSI document on dilution testing and are the same as those used for agar dilution tests. In fact, the broth microdilution method is now considered the international reference susceptibility testing method. This chapter focuses on in-house preparation and use of broth microdilution panels. Most of the principles and practices discussed in the chapter are pertinent to the broth microdilution method regardless of the source of the antibiotic panels. The dilution scheme for preparing broth microdilution panels is the same as that described for agar and broth macrodilution methods. The definitions of the interpretive categories and the comments concerning the use of the standards for agar and broth macrodilution methods are also applicable to broth microdilution methods. Like full-range dilution testing, breakpoint methods require the use of appropriately adjusted and supplemented Mueller-Hinton broth or agar. In addition, the standard inoculation, incubation, and interpretation procedures recommended for the full-range dilution methods should be followed.

Citation: Patel J, Tenover F, Turnidge J, Jorgensen J. 2011. Susceptibility Test Methods: Dilution and Disk Diffusion Methods, p 1122-1143. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch68

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Susceptibility Test Methods: Dilution and Disk Diffusion Methods
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Citation: Patel J, Tenover F, Turnidge J, Jorgensen J. 2011. Susceptibility Test Methods: Dilution and Disk Diffusion Methods, p 1122-1143. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch68
/content/10.1128/9781555816728.chap68.fig68-1
FIGURE 1

Example of the wild-type (WT) MIC distribution data that are used by EUCAST to set the epidemiological cutoff.

10.1128/9781555816728/fig68-1_thmb.gif
10.1128/9781555816728/fig68-1.gif
Image of FIGURE 1
FIGURE 1

Example of the wild-type (WT) MIC distribution data that are used by EUCAST to set the epidemiological cutoff.

Citation: Patel J, Tenover F, Turnidge J, Jorgensen J. 2011. Susceptibility Test Methods: Dilution and Disk Diffusion Methods, p 1122-1143. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch68
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References

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1. Andrews, J. M., for the BSAC Working Party on Susceptibility Testing. 2001. BSAC standardized disc susceptibility testing method. J. Antimicrob. Chemother. 48 (Suppl. 1): 43– 57.
2. Andrews, J. M. 2001. Determination of minimum inhibitory concentrations. J. Antimicrob. Chemother. 48 (Suppl. 1): 5– 16.
3. Andrews, J. M.,, F. J. Boswell,, and R. Wise. 2000. Evaluation of the Oxoid Aura image system for measuring zones of inhibition with the disc diffusion technique. J. Antimicrob. Chemother. 46: 535– 540.
4. Baker, C. N.,, S. A. Stocker,, D. H. Culver,, and C. Thornsberry. 1991. Comparison of the E Test to agar dilution, broth microdilution, and agar diffusion susceptibility testing techniques by using a special challenge set of bacteria. J. Clin. Microbiol. 29: 533– 538.
5. Barry, A. L.,, R. E. Badal,, and R. W. Hawkinson. 1983. Influence of inoculum growth phase on microdilution susceptibility tests. J. Clin. Microbiol. 18: 645– 651.
6. Barry, A. L.,, L. B. Reller,, G. H. Miller,, J. A. Washington,, F. D. Schoenknect,, L. R. Peterson,, R. S. Hare,, and C. Knapp. 1992. Revision of standards for adjusting the cation content of Mueller-Hinton broth for testing susceptibility of Pseudomonas aeruginosa to aminoglycosides. J. Clin. Microbiol. 30: 585– 589.
7. Bauer, A. W.,, W. M. Kirby,, J. C. Sherris,, and M. Turck. 1966. Antibiotic susceptibility testing by a standardized single disk method. Am. J. Clin. Pathol. 45: 493– 496.
8. Bauer, A. W.,, and J. C. Sherris. 1964. The determination of sulfonamide susceptibility of bacteria. Chemotherapy 15: 1– 19.
9. Bell, S. M. 1988. Additions and modifications to the range of antibiotics tested by the CDS method of antibiotic sensitivity testing. Pathology 20: 303– 304.
10. Bell, S. M. 1975. The CDS disc method of antibiotic sensitivity testing (calibrated dichotomous sensitivity test). Pathology 7( Suppl. 1): 1– 48.
11. Berke, I.,, and P. M. Tierno, Jr. 1996. Comparison of efficacy and cost-effectiveness of BIOMIC VIDEO and Vitek antimicrobial susceptibility test systems for use in the clinical microbiology laboratory. J. Clin. Microbiol. 34: 1980– 1984.
12. Brenner, V. C.,, and J. C. Sherris. 1972. Influence of different media and bloods on results of diffusion antibiotic susceptibility tests. Antimicrob. Agents Chemother. 1: 116– 122.
13.British Society for Antimicrobial Chemotherapy. 1991. A guide to sensitivity testing. Report of the Working Party on Antibiotic Sensitivity Testing of the British Society for Antimicrobial Chemotherapy. J. Antimicrob. Chemother. 27 (Suppl. D): 1– 50.
14. Brown, D. F. 1990. The comparative methods of antimicrobial susceptibility testing—time for a change? J. Antimicrob. Chemother. 25: 307– 310.
15. Brown, D. F. J. 1994. Developments in antimicrobial susceptibility testing. Rev. Med. Microbiol. 5: 65– 75.
16. Brunden, M. N.,, G. E. Zurenko,, and B. Kapik. 1992. Modification of the error-rate bounded classification scheme for use with two MIC break points. Diagn. Microbiol. Infect. Dis. 15: 135– 140.
17. Chaitram, J. M.,, L. A. Jevitt,, S. Lary,, and F. C. Tenover. 2003. The World Health Organization’s External Quality Assurance System Proficiency Testing Program has improved the accuracy of antimicrobial susceptibility testing and reporting among participating laboratories using NCCLS methods. J. Clin. Microbiol. 41: 2372– 2377.
18. Clinical and Laboratory Standards Institute. 2008. Development of In Vitro Susceptibility Testing Criteria and Quality Control Parameters. Approved guideline M23-A3. Clinical and Laboratory Standards Institute, Wayne, PA.
19. Clinical and Laboratory Standards Institute. 2006. Methods for Antimicrobial Dilution and Disk Susceptibility Testing of Infrequently Isolated or Fastidious Bacteria. Proposed guideline M45-P. Clinical and Laboratory Standards Institute, Wayne, PA.
20. Clinical and Laboratory Standards Institute. 2009. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically, 8th ed. Approved standard M7-A8. Clinical and Laboratory Standards Institute, Wayne, PA.
21. Clinical and Laboratory Standards Institute. 2009. Performance Standards for Antimicrobial Disk Susceptibility Tests, 10th ed. Approved standard M2-A10. Clinical and Laboratory Standards Institute, Wayne, PA.
22. Clinical and Laboratory Standards Institute. 2010. Performance Standards for Antimicrobial Susceptibility Testing. Nineteenth informational supplement. M100-S20. Clinical and Laboratory Standards Institute, Wayne, PA.
23. Clinical and Laboratory Standards Institute. 2006. Protocols for Evaluating Dehydrated Mueller-Hinton Agar. Approved standard M6-A2. Clinical and Laboratory Standards Institute, Wayne, PA
24. Clinical and Laboratory Standards Institute/NCCLS. 2001. Evaluation of Lots of Dehydrated Mueller-Hinton Broth for Antimicrobial Susceptibility Testing. Proposed guideline M23-P, vol. 21. NCCLS, Wayne, PA.
25. Coyle, M. B.,, M. F. Lampe,, C. L. Aitken,, P. Feigl,, and J. C. Sherris. 1976. Reproducibility of control strains for antibiotic susceptibility testing. Antimicrob. Agents Chemother. 10: 436– 440.
26. Daly, J. S.,, R. A. Dodge,, R. H. Glew,, D. T. Soja,, B. A. DeLuca,, and S. Hebert. 1997. Effect of zinc concentration in Mueller-Hinton agar on susceptibility of Pseudomonas aeruginosa to imipenem. J. Clin. Microbiol. 35: 1027– 1029.
27. D’Amato, R. F.,, C. Thornsberry,, C. N. Baker,, and L. A. Kirven. 1975. Effect of calcium and magnesium ions on the susceptibility of Pseudomonas species to tetracycline, gentamicin [sic] polymyxin B, and carbenicillin. Antimicrob. Agents Chemother. 7: 596– 600.
28. de Lencastre, H.,, A. M. Sa Figueiredo,, C. Urban,, J. Rahal,, and A. Tomasz. 1991. Multiple mechanisms of methicillin resistance and improved methods for detection in clinical isolates of Staphylococcus aureus. Antimicrob. Agents Chemother. 35: 632– 639.
29. Deutsches Institut für Normung. 2002. Susceptibility Testing of Pathogens to Antimicrobial Agents, part 3. Agar Diffusion Test, vol. DIN 58940-3. Beuth Verlag, Berlin, Germany.
30. Deutsches Institut für Normung. 2003. Susceptibility Testing of Pathogens to Antimicrobial Agents,part 6. Determination of the Minimum Inhibitory Concentration (MIC) with the Agar Dilution Method, vol. DIN 58940-6. Beuth Verlag, Berlin, Germany.
31. Diekema, D. J.,, K. Lee,, P. Raney,, L. A. Herwaldt,, G. V. Doern,, and F. C. Tenover. 2004. Accuracy and appropriateness of antimicrobial susceptibility test reporting for bacteria isolated from blood cultures. J. Clin. Microbiol. 42: 2258– 2260.
32. Doern, G. V. 1987. Breakpoint susceptibility testing. Clin. Microbiol. Newsl. 9: 81– 84.
33. Ericsson, H. M.,, and J. C. Sherris. 1971. Antibiotic sensitivity testing. Report of an international collaborative study. Acta Pathol. Microbiol. Scand. Sect. B Suppl. 217: 1– 90.
34.European Committee for Antimicrobial Susceptibility Testing. 2000. Determination of minimum inhibitory concentrations (MICs) of antibacterial agents by agar dilution. Clin. Microbiol. Infect. 6: 509– 515.
35. Ferraro, M. J. 2001. Should we reevaluate antibiotic breakpoints? Clin. Infect. Dis. 33( Suppl. 3): S227– S229.
36. Fiebelkorn, K. R.,, S. A. Crawford,, M. L. McElmeel,, and J. H. Jorgensen. 2003. Practical disk diffusion method for detection of inducible clindamycin resistance in Staphylococcus aureus and coagulase-negative staphylococci. J. Clin. Microbiol. 41: 4740– 4744.
37. Fuchs, P. C.,, A. L. Barry,, and S. D. Brown. 2000. Daptomycin susceptibility tests: interpretive criteria, quality control, and effect of calcium on in vitro tests. Diagn. Microbiol. Infect. Dis. 38: 51– 58.
38. Gales, A. C.,, A. O. Reis,, and R. N. Jones. 2001. Contemporary assessment of antimicrobial susceptibility testing methods for polymyxin B and colistin: review of available interpretative criteria and quality control guidelines. J. Clin. Microbiol. 39: 183– 190.
39. Hayden, M. K.,, K. Rezai,, R. A. Hayes,, K. Lolans,, J. P. Quinn,, and R. A. Weinstein. 2005. Development of daptomycin resistance in vivo in methicillin-resistant Staphylococcus aureus. J. Clin. Microbiol. 43: 5285– 5287.
40. Health Care Financing Administration. 1992. Clinical Laboratory Improvement Amendments of 1988, final rule. Fed. Regist. 57: 7001– 7186.
41. Huang, M. B.,, C. N. Baker,, S. Banerjee,, and F. C. Tenover. 1992. Accuracy of the E test for determining antimicrobial susceptibilities of staphylococci, enterococci, Campylobacter jejuni, and gram-negative bacteria resistant to antimicrobial agents. J. Clin. Microbiol. 30: 3243– 3248.
42. Huang, M. B.,, T. E. Gay,, C. N. Baker,, S. N. Banerjee,, and F. C. Tenover. 1993. Two percent sodium chloride is required for susceptibility testing of staphylococci with oxacillin when using agar-based dilution methods. J. Clin. Microbiol. 31: 2683– 2688.
43. International Organizationfor Standardization. 2006. Susceptibility Testing of Infectious Agents and Evaluation of Performance of Antimicrobial Susceptibility Devices, part 1. Reference Method for Testing the In Vitro Activity of Antimicrobial Agents against Bacteria Involved in Infectious Diseases, vol. ISO/DIS 20776-1. International Organization for Standardization, Geneva, Switzerland.
44. International Organization for Standardization. 2006. Susceptibility Testing of Infectious Agents and Evalutaion of Performance of Antimicrobial Susceptibility Devices, part 2. Evaluation of Performance of Antimicrobial Susceptibility Test Devices, vol. ISO/DIS 20776-2. International Organization for Standardization, Geneva, Switzerland.
45. Japanese Society for Chemotherapy. 1993. Report of the Committee for Japanese Standards for Antimicrobial Susceptibility Testing for Bacteria. Chemotherapy 41: 183– 189.
46. Japanese Society for Chemotherapy. 1990. Report of the Committee for Japanese Standards for Antimicrobial Susceptibility Testing for Bacteria. Chemotherapy 38: 102– 105.
47. Jevitt, L. A.,, G. M. Thorne,, M. M. Traczewski,, R. N. Jones,, J. E. McGowan, Jr.,, F. C. Tenover,, and S. D. Brown. 2006. Multicenter evaluation of the Etest and disk diffusion methods for differentiating daptomycin-susceptible from nondaptomycin-susceptible Staphylococcus aureus isolates. J. Clin. Microbiol. 44: 3098– 3104.
48. Jorgensen, J. H. 1993. Selection criteria for an antimicrobial susceptibility testing system. J. Clin. Microbiol. 31: 2841– 2844.
49. Jorgensen, J. H. 1993. Selection of antimicrobial agents for routine testing in a clinical microbiology laboratory. Diagn. Microbiol. Infect. Dis. 16: 245– 249.
50. Jorgensen, J. H.,, and M. J. Ferraro. 1998. Antimicrobial susceptibility testing: general principles and contemporary practices. Clin. Infect. Dis. 26: 973– 980.
51. Jorgensen, J. H.,, J. M. Swenson,, F. C. Tenover,, M. J. Ferraro,, J. A. Hindler,, and P. R. Murray. 1994. Development of interpretive criteria and quality control limits for broth microdilution and disk diffusion antimicrobial susceptibility testing of Streptococcus pneumoniae. J. Clin. Microbiol. 32: 2448– 2459.
52. Kahlmeter, G.,, D. F. Brown,, F. W. Goldstein,, A. P. MacGowan,, J. W. Mouton,, I. Odenholt,, A. Rodloff,, C. J. Soussy,, M. Steinbakk,, F. Soriano,, and O. Stetsiouk. 2006. European Committee on Antimicrobial Susceptibility Testing (EUCAST) technical notes on antimicrobial susceptibility testing. Clin. Microbiol. Infect. 12: 501– 503.
53. King, A.,, and D. F. Brown. 2001. Quality assurance of antimicrobial susceptibility testing by disc diffusion. J. Antimicrob. Chemother. 48 (Suppl. 1): 71– 76.
54. Lewis, J. S., II,, and J. H. Jorgensen. 2005. Inducible clindamycin resistance in staphylococci: should clinicians and microbiologists be concerned? Clin. Infect. Dis. 40: 280– 285.
55. Lewis, J. S., II,, A. Owens,, J. Cadena,, K. Sabol,, J. E. Patterson,, and J. H. Jorgensen. 2005. Emergence of daptomycin resistance in Enterococcus faecium during daptomycin therapy. Antimicrob. Agents Chemother. 49: 1664– 1665.
56. Li, X. Z. 2005. Quinolone resistance in bacteria: emphasis on plasmid-mediated mechanisms. Int. J. Antimicrob. Agents 25: 453– 463.
57. MacGowan, A. P.,, and R. Wise. 2001. Establishing MIC breakpoints and the interpretation of in vitro susceptibility tests. J. Antimicrob. Chemother. 48 (Suppl. 1): 17– 28.
58. McDermott, S. N.,, and T. F. Hartley. 1989. New datum handling methods for the quality control of antibiotic solutions and plates used in the antimicrobial susceptibility test. J. Clin. Microbiol. 27: 1814– 1825.
59. Medeiros, A. A.,, and J. Crellin. 2000. Evaluation of the Sirscan automated zone reader in a clinical microbiology laboratory. J. Clin. Microbiol. 38: 1688– 1693.
60. Members of the SFM Antibiogram Committee. 2003. Comite de l’Antibiogramme de la Societe Francaise de Microbiologie report 2003. Int. J. Antimicrob. Agents 21: 364– 391.
61. Metzler, C. M.,, and R. M. DeHaan. 1974. Susceptibility tests of anaerobic bacteria: statistical and clinical considerations. J. Infect. Dis. 130: 588– 594.
62. Olsson-Liljequist, B.,, P. Larsson,, M. Walder,, and H. Miorner. 1997. Antimicrobial susceptibility testing in Sweden. III. Methodology for susceptibility testing. Scand. J. Infect. Dis. Suppl. 105: 13– 23.
63. Pfaller, M. A.,, and R. N. Jones. 2006. Performance accuracy of antibacterial and antifungal susceptibility test methods: report from the College of American Pathologists Microbiology Surveys Program (2001-2003). Arch. Pathol. Lab. Med. 130: 767– 778.
64. Queenan, A. M.,, and K. Bush. 2007. Carbapenemases: the versatile β-lactamases. Clin. Microbiol. Rev. 20: 440– 458.
65. Ringertz, S.,, B. Olsson-Liljequist,, G. Kahlmeter,, and G. Kronvall. 1997. Antimicrobial susceptibility testing in Sweden. II. Species-related zone diameter breakpoints to avoid interpretive errors and guard against unrecognized evolution of resistance. Scand. J. Infect. Dis. Suppl. 105: 8– 12.
66. Rosenberg, J.,, F. C. Tenover,, J. Wong,, W. Jarvis,, and D. J. Vugia. 1997. Are clinical laboratories in California accurately reporting vancomycin-resistant enterococci? J. Clin. Microbiol. 35: 2526– 2530.
67. Sahm, D. F.,, C. N. Baker,, R. N. Jones,, and C. Thornsberry. 1984. Influence of growth medium on the in vitro activities of second and third generation cephalosporins against Streptococcus faecalis. J. Clin. Microbiol. 20: 561– 567.
68. Steers, E.,, E. L. Foltz,, B. S. Graves,, and H. J. Suriano. 1959. Comparison of bacterial susceptibility to antibiotics as determined by the plate dilution method and by the disc method. Antibiot. Annu. 7: 604– 613.
69.Swedish Reference Group for Antibiotics. 1981. A revised system for antibiotic sensitivity testing. Scand. J. Infect. Dis. 13: 148– 152.
70. Swenson, J. M.,, K. F. Anderson,, D. R. Lonsway,, A. Thompson,, S. K. McAllister,, B. M. Limbago,, R. B. Carey,, F. C. Tenover,, and J. B. Patel. 2009. Accuracy of commercial and reference susceptibility testing methods for detecting vancomycin-intermediate Staphylococcus aureus. J. Clin. Microbiol. 47: 2013– 2017.
71. Swenson, J. M.,, W. B. Brasso,, M. J. Ferraro,, D. J. Hardy,, C. C. Knapp,, D. Lonsway,, S. McAllister,, L. B. Reller,, H. S. Sader,, D. Shortridge,, R. Skov,, M. P. Weinstein,, B. L. Zimmer,, and J. B. Patel. 2009. Correlation of cefoxitin MICs with the presence of mecA in Staphylococcus spp. J. Clin. Microbiol. 47: 1902– 1905.
72. Swenson, J. M.,, and F. C. Tenover. 2005. Results of disk diffusion testing with cefoxitin correlate with presence of mecA in Staphylococcus spp. J. Clin. Microbiol. 43: 3818– 3823.
73. Tenover, F. C.,, M. V. Lancaster,, B. C. Hill,, C. D. Steward,, S. A. Stocker,, G. A. Hancock,, C. M. O’Hara,, S. K. McAllister,, N. C. Clark,, and K. Hiramatsu. 1998. Characterization of staphylococci with reduced susceptibilities to vancomycin and other glycopeptides. J. Clin. Microbiol. 36: 1020– 1027.
74. Tenover, F. C.,, and L. C. McDonald. 2005. Vancomycinresistant staphylococci and enterococci: epidemiology and control. Curr. Opin. Infect. Dis. 18: 300– 305.
75. Tenover, F. C.,, J. M. Swenson,, C. M. O’Hara,, and S. A. Stocker. 1995. Ability of commercial and reference antimicrobial susceptibility testing methods to detect vancomycin resistance in enterococci. J. Clin. Microbiol. 33: 1524– 1527.
76. Thornsberry, C.,, and L. K. McDougal. 1983. Successful use of broth microdilution in susceptibility tests for methicillinresistant (heteroresistant) staphylococci. J. Clin. Microbiol. 18: 1084– 1091.
77. Toohey, M.,, G. Francis,, and N. Stingemore. 1990. Variation in Iso-Sensitest agar affecting β-lactam testing. Newsl. Antimicrob. Spec. Interest Group Aust. Soc. Microbiol. 6: 6– 8.
78. Turnidge, J.,, G. Kahlmeter,, and G. Kronvall. 2006. Statistical characterisation of bacterial wild-type MIC value distributions and the determination of epidemiological cut-off values. Clin. Microbiol. Infect. 12: 418– 425.
79. Turnidge, J. D.,, and J. M. Bell,. 2005. Antimicrobial susceptibility on solid media, p. 8– 60. In V. Lorien (ed.), Antibiotics in Laboratory Medicine, 5th ed. Lippincott Williams & Wilkins, Philadelphia, PA.
80. Ward, P.,, S. Palladino,, B. McLaren,, R. J. Rathur,, and J. C. Looker. 1993. The effect of increased agar concentration in susceptibility testing media on MICs of antimicrobials for gramnegative bacilli. J. Antimicrob. Chemother. 31: 1005– 1007.
81. Ward, P. B.,, S. Palladino,, J. C. Looker,, and P. Feddema. 1993. p-Nitrophenylglycerol in susceptibility testing media alters the MICs of antimicrobials for Pseudomonas aeruginosa. J. Antimicrob. Chemother. 31: 489– 496.
82. Wheat, P. F. 1989. The agar-dilution susceptibility technique: past and present. Clin. Microbio. Newsl. 11: 164– 166.
83. Williams, J. D. 1990. Prospects for standardisation of methods and guidelines for disc susceptibility testing. Eur. J. Clin. Microbiol. Infect. Dis. 9: 496– 501.

Tables

Susceptibility Test Methods: Dilution and Disk Diffusion Methods
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Citation: Patel J, Tenover F, Turnidge J, Jorgensen J. 2011. Susceptibility Test Methods: Dilution and Disk Diffusion Methods, p 1122-1143. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch68
/content/10.1128/9781555816728.chap68.tab68-1
TABLE 1

Antibacterial susceptibility results that may be extrapolated from other test results

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10.1128/9781555816728/tab68-1.gif
Generic image for table
TABLE 1

Antibacterial susceptibility results that may be extrapolated from other test results

Citation: Patel J, Tenover F, Turnidge J, Jorgensen J. 2011. Susceptibility Test Methods: Dilution and Disk Diffusion Methods, p 1122-1143. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch68
/content/10.1128/9781555816728.chap68.tab68-2
TABLE 2

Antimicrobial agents recommended for routine dilution and disk diffusion susceptibility testing

Modified from CLSI document M100-S20 ( ) with permission. Current standards and supplements to them may be obtained from the CLSI, 940 West Valley Rd., Suite 1400, Wayne, PA 19087-1898.

Group A comprises primary drugs to be tested and reported, group B comprises those to be tested as primary drugs but reported selectively, group C comprises supplemental drugs to be reported selectively, and group U comprises drugs to be tested with urinary isolates only

Non- include spp. and other nonfastidious, non-glucose-fermenting gram-negative bacilli but exclude , spp., , , , and , since there are separate listings of suggested drugs to test and report for them.

Those agents marked with an asterisk should be tested only by using an MIC method and not by using disk diffusion.

Results of tests with penicillin apply to other penicillins (e.g., ampicillin, amoxicillin, carboxypenicillins, and ureidopenicillins) against β-lactamase-negative enterococci.

Combination therapy consisting of penicillin, ampicillin, or vancomycin and an aminoglycoside is recommended for serious infections.

Staphylococci resistant to the penicillinase-resistant penicillins should also be considered resistant to penicillins, β-lactam–β-lactamase-inactivating combinations, cephalosporins (except cephalosporins with anti-MRSA activity), and carbapenems.

Cephalothin test results should only be used to represent oral agents, i.e., cefadroxil, cefpodoxime, cephalexin, and loracarbef. Older data which suggest that cephalothin results could predict susceptibility to some other cephalosporins may still be correct, but there are no recent data to confirm this.

Cefoxitin is used as a surrogate for oxacillin for staphylococci.

For use of aminoglycosides to screen enterococci for synergy resistance, see the sections “Breakpoint Susceptibility Tests” and “Resistance Screens.”

Doxycycline or minocycline may be tested on a supplemental basis because of their greater activities against some nonfermentative gram-negative bacilli and staphylococci.

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TABLE 2

Antimicrobial agents recommended for routine dilution and disk diffusion susceptibility testing

Modified from CLSI document M100-S20 ( ) with permission. Current standards and supplements to them may be obtained from the CLSI, 940 West Valley Rd., Suite 1400, Wayne, PA 19087-1898.

Group A comprises primary drugs to be tested and reported, group B comprises those to be tested as primary drugs but reported selectively, group C comprises supplemental drugs to be reported selectively, and group U comprises drugs to be tested with urinary isolates only

Non- include spp. and other nonfastidious, non-glucose-fermenting gram-negative bacilli but exclude , spp., , , , and , since there are separate listings of suggested drugs to test and report for them.

Those agents marked with an asterisk should be tested only by using an MIC method and not by using disk diffusion.

Results of tests with penicillin apply to other penicillins (e.g., ampicillin, amoxicillin, carboxypenicillins, and ureidopenicillins) against β-lactamase-negative enterococci.

Combination therapy consisting of penicillin, ampicillin, or vancomycin and an aminoglycoside is recommended for serious infections.

Staphylococci resistant to the penicillinase-resistant penicillins should also be considered resistant to penicillins, β-lactam–β-lactamase-inactivating combinations, cephalosporins (except cephalosporins with anti-MRSA activity), and carbapenems.

Cephalothin test results should only be used to represent oral agents, i.e., cefadroxil, cefpodoxime, cephalexin, and loracarbef. Older data which suggest that cephalothin results could predict susceptibility to some other cephalosporins may still be correct, but there are no recent data to confirm this.

Cefoxitin is used as a surrogate for oxacillin for staphylococci.

For use of aminoglycosides to screen enterococci for synergy resistance, see the sections “Breakpoint Susceptibility Tests” and “Resistance Screens.”

Doxycycline or minocycline may be tested on a supplemental basis because of their greater activities against some nonfermentative gram-negative bacilli and staphylococci.

Citation: Patel J, Tenover F, Turnidge J, Jorgensen J. 2011. Susceptibility Test Methods: Dilution and Disk Diffusion Methods, p 1122-1143. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch68
/content/10.1128/9781555816728.chap68.tab68-3
TABLE 3

Interpretive standards for dilution and disk diffusion susceptibility testing

Adapted from CLSI data ( ) with permission. The interpretive data are valid only if the methodologies in documents M2-A10 ( ) and M7-A8 ( ) are followed. Breakpoints for some agents apply only to certain genera or species. The CLSI frequently updates the interpretive tables through new editions of the standards and supplements to them. Users should refer to the most recent editions. The current standards and supplements to them may be obtained from the CLSI, 940 West Valley Rd., Suite 1400, Wayne, PA 19087-1898.

Penicillin should be used as the class representative for all penicillins (e.g., ampicillin, amoxicillin, mezlocillin, piperacillin, and ticarcillin). Isolates for which MICs are ≤0.03 mg of penicillin per ml generally do not produce β-lactamase, whereas those for which MICs are ≥0.25 mg/ml do and should be regarded as resistant to penicillins. Isolates for which MICs of penicillin are 0.06 or 0.12 mg/ml should be tested for β-lactamase.

Therapy for serious enterococcal infections requires high doses of penicillin or ampicillin in combination with an aminoglycoside. Vancomycin may be substituted for the penicillin in instances of penicillin hypersensitivity or of penicillin or ampicillin resistance.

Oxacillin or methicillin may be tested; however, oxacillin is preferred because of its greater stability in vitro. The results from the testing of oxacillin apply also to other penicillinase-resistant penicillins. Oxacillin-resistant staphylococci should be considered resistant to all penicillins, cephalosporins (except cephalosporins with anti-MRSA activity), carbacephems, carbapenems, and β-lactam–β-lactamase inhibitor combinations. Disk testing with cefoxitin is the most sensitive and specific method for phenotypic detection of A-mediated oxacillin resistance in staphylococci ( ).

For the treatment of urinary tract infections only.

Interpretive criteria are from the Food and Drug Administration drug label.

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TABLE 3

Interpretive standards for dilution and disk diffusion susceptibility testing

Adapted from CLSI data ( ) with permission. The interpretive data are valid only if the methodologies in documents M2-A10 ( ) and M7-A8 ( ) are followed. Breakpoints for some agents apply only to certain genera or species. The CLSI frequently updates the interpretive tables through new editions of the standards and supplements to them. Users should refer to the most recent editions. The current standards and supplements to them may be obtained from the CLSI, 940 West Valley Rd., Suite 1400, Wayne, PA 19087-1898.

Penicillin should be used as the class representative for all penicillins (e.g., ampicillin, amoxicillin, mezlocillin, piperacillin, and ticarcillin). Isolates for which MICs are ≤0.03 mg of penicillin per ml generally do not produce β-lactamase, whereas those for which MICs are ≥0.25 mg/ml do and should be regarded as resistant to penicillins. Isolates for which MICs of penicillin are 0.06 or 0.12 mg/ml should be tested for β-lactamase.

Therapy for serious enterococcal infections requires high doses of penicillin or ampicillin in combination with an aminoglycoside. Vancomycin may be substituted for the penicillin in instances of penicillin hypersensitivity or of penicillin or ampicillin resistance.

Oxacillin or methicillin may be tested; however, oxacillin is preferred because of its greater stability in vitro. The results from the testing of oxacillin apply also to other penicillinase-resistant penicillins. Oxacillin-resistant staphylococci should be considered resistant to all penicillins, cephalosporins (except cephalosporins with anti-MRSA activity), carbacephems, carbapenems, and β-lactam–β-lactamase inhibitor combinations. Disk testing with cefoxitin is the most sensitive and specific method for phenotypic detection of A-mediated oxacillin resistance in staphylococci ( ).

For the treatment of urinary tract infections only.

Interpretive criteria are from the Food and Drug Administration drug label.

Citation: Patel J, Tenover F, Turnidge J, Jorgensen J. 2011. Susceptibility Test Methods: Dilution and Disk Diffusion Methods, p 1122-1143. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch68
/content/10.1128/9781555816728.chap68.tab68-4
TABLE 4

Non-U.S. disk diffusion methods for susceptibility testing

http://www.eucast.org.

http://www.bsac.org.uk.

MH, Mueller-Hinton agar.

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Generic image for table
TABLE 4

Non-U.S. disk diffusion methods for susceptibility testing

http://www.eucast.org.

http://www.bsac.org.uk.

MH, Mueller-Hinton agar.

Citation: Patel J, Tenover F, Turnidge J, Jorgensen J. 2011. Susceptibility Test Methods: Dilution and Disk Diffusion Methods, p 1122-1143. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch68

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