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Chapter 7.6 : Procedures for Identification from Culture

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Abstract:

Whenever possible, mycobacteria should be identified to the species level. A level laboratory must routinely process and culture at least 20 colonial specimens per week in order to ensure proficiency in identifying However, this low workload necessitates referring nontuberculosis mycobacteria to a level III laboratory for identification ( ). In addition to colonial morphology ( ) and acid-fastness, the identification of mycobacteria is largely based on rapid DNA probes (AccuProbe) and conventional methods. The conventional biochemicals used to identify mycobacteria are discussed in this procedure.

Citation: Garcia L. 2010. Procedures for Identification from Culture, p 329-346. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch7.6
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Figures

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Figure 7.6.1-1

Procedure for identification of acid-fast bacilli from culture ( ). HPLC, high-performance liquid chromatography.

Citation: Garcia L. 2010. Procedures for Identification from Culture, p 329-346. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch7.6
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References

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1.California State Department of Public Health, Department of Mycobacteriology. Procedures. California State Department of Public Health, Berkeley.
2. Della-Latta, P.,, and I. Weitzman,. 1998. Identification procedures from culture, p. 187197. In H. D. Isenberg (ed.), Essential Procedures for Clinical Microbiology. ASM Press, Washington, D.C..
3. Kent, P. T.,, and G. P. Kubica. 1985. Public Health Mycobacteriology. A Guide for the Level III Laboratory. U.S. Department of Health and Human Services, Centers for Disease Control, Atlanta, Ga..
4. Kubica, G.,, and L. Wayne. 1984. The Mycobacteria, a Sourcebook. Marcel Dekker, Inc., New York, N.Y..
5. Lutz, B., 1992. Identification tests for mycobacteria, p. 3.12.13.12.28. In H. D. Isenberg (ed.), Clinical Microbiology Procedures Handbook, vol. 1. American Society for Microbiology, Washington, D.C..
6. Pfyffer, G. E.,, B. A. Brown-Elliott,, and R. J. Wallace, Jr., 2003. Mycobacterium, p. 532584. In P. R. Murray,, E. J. Baron,, J. H. Jorgensen,, M. A. Pfaller,, and R. H. Yolken (ed.), Manual of Clinical Microbiology, 8th ed. ASM Press, Washington, D.C..
7. Roberts, G. D.,, E. W. Koneman,, and Y. K. Kim,. 1991. Mycobacterium, p. 304339. In A. Balows,, W. J. Hausler, Jr.,, K. L. Herrmann,, H. D. Isenberg,, and H. J. Shadomy (ed.), Manual of Clinical Microbiology, 5th ed. American Society for Microbiology, Washington, D.C..
8. Della-Latta, P. 1996. Workflow and optimal protocols for laboratories in industrialized countries. Clin. Lab. Med. 16:677695.
9. Orange County Public Health Department Laboratory, Mycobacteriology/Mycology Section. Standard Operating Procedures for the Mycobacteriology Laboratory. Orange County Public Health Department Laboratory, Santa Ana, Calif..
10. Silcox, V. A., 1992. Identification of mycobacteria, p. 3.11.13.11.11. In H. D. Isenberg (ed.), Clinical Microbiology Procedures Handbook, vol. 1. American Society for Microbiology, Washington, D.C..
1. Ellner, P. D.,, T. E. Kiehn,, R. Cammarata,, and M. Hosmer. 1988. Rapid detection and identification of pathogenic mycobacteria by combining radiometric and nucleic acid probe methods. J. Clin. Microbiol. 26:13491352.
2.Gen-Probe Inc. 1991. AccuProbe Mycobacterium tuberculosis Complex Culture Identification Test. Gen-Probe Inc., San Diego, Calif..
3. Gonzalez, R.,, and B. A. Hanna. 1987. Evaluation of Gen-Probe DNA hybridization systems for the identification of M. tuberculosis and M. avium-intracellulare. Diagn. Microbiol. Infect. Dis. 8:6977.
4. Kaminski, D. A.,, and D. J. Hardy. 1995. Selective utilization of DNA probes for identification of Mycobacterium species on the basis of cord formation in primary BACTEC 12B cultures. J. Clin. Microbiol. 33:15481550.
1. Laszlo, A.,, and S. H. Siddiqi. 1984. Evaluation of a rapid radiometric differentiation test for the Mycobacterium tuberculosis complex by selective inhibition with p-nitro-α-acetylamino-b-hydroxy-propiophenone. J. Clin. Microbiol. 19:694695.
2. Morgan, M. A.,, K. A. Doerr,, H. O. Hempel,, N. M. L. Goodman,, and G. D. Roberts. 1985. Evaluation of the p-nitro-α-acetylamino-b-hydroxy-propiophenone differential test for identification of Mycobacterium tuberculosis complex. J. Clin. Microbiol. 21:634635.
3. Siddiqi, S. H.,, C. C. Hwangbo,, V. Silcox,, R. C. Good,, D. E. Sneider, Jr., and G. Middlebrook. 1984. Rapid radiometric methods to detect and differentiate M. tuberculosis/M. bovis from other mycobacterial species. Am. Rev. Respir. Dis. 130:634640.

Tables

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Table 7.6.1-1

Preparation of arylsulfatase standards ( )

Citation: Garcia L. 2010. Procedures for Identification from Culture, p 329-346. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch7.6
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Table 7.6.1-2

Distinctive properties of cultivable mycobacteria encountered in clinical specimens

Modified from reference . Plus and minus signs indicate the presence and absence, respectively, of the feature; blank spaces indicate either that the information is not currently available or that the property is unimportant. V, variable; ±, usually present; −/+, usually absent. The percentage of CDC-tested strains positive in each test is given in parentheses, and the test result is based on these percentages.

R, rough; S, smooth; SR, intermediate in roughness; t, thin or transparent; f, filamentous extensions.

P, photochromogenic; S, scotochromogenic; N, nonchromogenic ( is scotochromogenic at 37°C and photochromogenic at 24°C).

Urease test performed by the method of Steadham (. 10:134–137, 1979).

Probe identifies complex.

Requires hemin as growth factor.

Arylsulfatase reaction at 14 days is positive.

Young cultures may be nonchromogenic or possess only pale pigment that may intensify with age.

Includes third biovariant complex.

Citation: Garcia L. 2010. Procedures for Identification from Culture, p 329-346. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch7.6
Generic image for table
Table 7.6.1-3

Features useful for making a presumptive identification of mycobacteria ( )

Citation: Garcia L. 2010. Procedures for Identification from Culture, p 329-346. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch7.6
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Citation: Garcia L. 2010. Procedures for Identification from Culture, p 329-346. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch7.6

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