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Chapter 5.11 : Serum Inhibitory and Bactericidal Titers

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Abstract:

Schlichter and MacLean first used serum inhibitory titers (SIT) to assess the effectiveness of penicillin in the treatment of subacute bacterial endocarditis ( ). Other recognized clinical indications for obtaining serum bactericidal titers (SBT) to monitor effective therapy are osteomyelitis, closed-space infections such as meningitis or joint infection, and the use of oral antimicrobial agents after intravenous therapy. Controversy surrounds and will continue to surround the methodology, and there are few clinical situations in which the test is indicated.

Citation: Garcia L. 2010. Serum Inhibitory and Bactericidal Titers, p 124-139. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch5.11
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Figures

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Use one plate for each patient, and test each sample in duplicate. Row: a horizontal line of 12 wells, designated by a letter. Column: a vertical line of 8 wells, designated by a number. Well: each well has a specific letter and number corresponding to row and column location. Symbols: , trough dilution rows; , peak dilution rows; , no-growth control wells; , growth control wells; □, empty wells.

Citation: Garcia L. 2010. Serum Inhibitory and Bactericidal Titers, p 124-139. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch5.11
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Image of Serum Inhibitory and Bactericidal Titer Worksheet
Serum Inhibitory and Bactericidal Titer Worksheet

cc = colony count determined from inoculum count verification plates

“Row” relates to microdilution tray (duplicate testing)

Citation: Garcia L. 2010. Serum Inhibitory and Bactericidal Titers, p 124-139. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch5.11
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Download as Powerpoint

References

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1.NCCLS. 1999. Methodology for the Serum Bactericidal Test. Approved guideline M21-A. NCCLS, Wayne, Pa.
2. Schlichter, J. G.,, and H. MacLean. 1947. A method for determining the effective therapeutic level in the treatment of subacute bacterial endocarditis with penicillin: a preliminary re-port. Am. Heart J. 34:209211.
3. Amsterdam, D., 1996. Susceptibility testing of antimicrobials in liquid media, p. 103105. In V. Lorian (ed.), Antibiotics in Laboratory Medicine, 4th ed. The Williams & Wilkins Co., Baltimore, Md.
4. Leggett, J. E.,, S. A. Wolz,, and W. A. Craig. 1989. Use of serum ultrafiltrate in the serum bactericidal test. J. Infect. Dis. 160:616623.
5. Stratton, C. W. 1988. Serum bactericidal test. Clin. Microbiol. Rev. 1:1926.
6. Taylor, P. C.,, F. D. Schoenknecht,, J. C. Sherris,, and E. C. Linner. 1983. Determination of minimal bactericidal concentration of oxacillin for Staphylococcus aureus: influence and significance of technical factors. Antimicrob. Agents Chemother. 23:142150.
7. Weinstein, M. P.,, C. W. Stratton,, A. Ackley,, H. B. Hawley,, B. D. Fisher,, D. V. Alcid,, D. S. Stephens,, and L. B. Reller. 1985. Multicenter collaborative evaluation of a standardized serum bactericidal test as a prognostic indicator in infective endocarditis. Am. J. Med. 78:262269.
8. Weinstein, M. P.,, C. W. Stratton,, H. B. Hawley,, A. Ackley,, and L. B. Reller. 1987. Multicenter collaborative evaluation of a standardized serum bactericidal test as a predictor of therapeutic osteomyelitis. Am. J. Med. 83:218222.
1.NCCLS. 1999. Methodology for the Serum Bactericidal Test. Approved guideline M21-A. NCCLS, Wayne, Pa.

Tables

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Citation: Garcia L. 2010. Serum Inhibitory and Bactericidal Titers, p 124-139. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch5.11
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SIT and SBT test conditions and media for various bacteria

Inhibitory portion of test (SIT) is incubated for 24 h. The information here applies to microdilution and macrodilution methods.

Specified broth is appropriate for growing inoculum to log phase also. Broth supplemented with NHS may be used ( Appendix 5.11–1 ). HTM, test medium; LHB, lysed horse blood; THB, Todd-Hewitt broth.

All incubations are at 35°C.

Citation: Garcia L. 2010. Serum Inhibitory and Bactericidal Titers, p 124-139. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch5.11
Generic image for table

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

When the sum of colonies from duplicate samples was equal to or less than the rejection value, the antimicrobial agent was declared lethal (a 0.999 or greater reduction in the final inoculum). (Adapted from 1980. Method for reliable determination of minimal lethal antibiotic concentrations. 699–708.) A 5% error (pipette error plus full sampling error for determination of final inoculum) was used. Error was based on duplicate samples for determination of the final inoculum size.

As determined from colony count plate.

Number of colonies.

Sensitivity and specificity were calculated for each specific final inoculum concentration and rejection value.

Citation: Garcia L. 2010. Serum Inhibitory and Bactericidal Titers, p 124-139. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch5.11

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