1887

Chapter 12.3 : Molecular Methods for Identification of Cultured Microorganisms

MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.

Preview this chapter:
Zoom in
Zoomout

Molecular Methods for Identification of Cultured Microorganisms, Page 1 of 2

| /docserver/preview/fulltext/10.1128/9781555817435/9781555815271_Chap12_3-1.gif /docserver/preview/fulltext/10.1128/9781555817435/9781555815271_Chap12_3-2.gif

Abstract:

Bacteria and fungi can be identified by using nucleic acid hybridization techniques. Nucleic acid probes can be used to confirm the identification of culture isolates that have been tested by presumptive identification methods. Alternatively, probes can be used as the primary method for identifying isolated organisms. Culture identification by probe hybridization is not dependent on the ability to detect minute quantities of nucleic acid, and thus sensitivity is not a limiting factor in this application of molecular technology. The advantage of probe-based identification is greatest for slow-growing organisms like the mycobacteria or for organisms for which convenient commercial identification systems are not available. Although the specificity of the available commercial probes is high and they facilitate rapid identification of a number of pathogens (Table 12.1-3), misidentifications do occur and serve to emphasize the need for caution in using any single characteristic in identification of a species. It should also be emphasized that at this point identification by probe hybridization is more expensive than conventional techniques for many organisms.

Citation: Garcia L. 2010. Molecular Methods for Identification of Cultured Microorganisms, p 366-379. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch12.3
Highlighted Text: Show | Hide
Loading full text...

Full text loading...

References

/content/book/10.1128/9781555817435.chap12.3
1. Cormican, M. G.,, and M. A. Pfaller,. 2001. Molecular pathology of infectious diseases, p. 12411253. In. J. B. Henry (ed.), Clinical Diagnosis and Management by Laboratory Methods, 20th ed. W. B. Saunders Company, Philadelphia, PA.
2. Nolte, F. S.,, and A. M. Caliendo,. 2003. Molecular detection and identification of microorganisms, p. 234256. 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, DC.
3. Tenover, F. C. 2007. Rapid detection and identification of bacterial pathogens using novel molecular technologies: infection control and beyond. Clin. Infect. Dis. 44:418423.
4. Woods, G. L. 2001. Molecular techniques in mycobacterial detection. Arch. Pathol. Lab. Med. 125:122126.
1.CLSI. 2004. Quality Assurance for Commercially Prepared Microbiological Culture Media, 3rd ed. Approved standard M22-A3. CLSI, Wayne, PA.
6. Daily, J. A.,, N. L. Clifton,, K. C. Seskin,, and W. M. Gooch III. 1991. Use of rapid, nonradioactive DNA probes in culture confirmation tests to detect Streptococcus agalactiae, Haemophilus influenzae, and Enterococcus spp. from pediatric patients with significant infections. J. Clin. Microbiol. 29:8082.
7. Denys, G. A.,, and R. B. Carey. 1992. Identification of Streptococcus pneumoniae with a DNA probe. J. Clin. Microbiol. 30:27252727.
8. Gen-Probe, Inc. 1995. AccuProbe culture identification test, package insert. Gen-Probe, Inc., San Diego, CA.
9. Kohne, D. E., 1990. The use of DNA probes to detect and identify microorganisms, p. 1135. In B. Kleger et al. (ed.), Rapid Methods in Clinical Microbiology. Plenum Press, New York, NY.
10. Lewis, J. S.,, D. Krnaig-Brown,, and D. A. Trainor. 1990. DNA probe confirmatory test for Neisseria gonorrhoeae. J. Clin. Microbiol. 28:23492350.
11. Stockman, K.,, K. A. Clark,, J. M. Hunt,, and G. D. Roberts. 1993. Evaluation of commercially available acridinium ester-labeled chemiluminescent DNA probes for culture identification of Blastomyces dermatitidis, Coccidioides immitis, Cryptococcus neoformans, and Histoplasma capsulatum. J. Clin. Microbiol. 31:845850.
1.. CLSI. 2004. Quality Assurance for Commercially Prepared Microbiological Culture Media, 3rd ed. Approved standard M22-A3. CLSI, Wayne, PA.
13. Badak, F. Z.,, S. Goksel,, and R. Sertoz. 1999. Use of nucleic acid probes for identification of Mycobacterium tuberculosis directly from MB/ BacT bottles. J. Clin. Microbiol. 37:16021605.
14. Devallois, A.,, K. S. Goh,, and N. Rastogi. 1997. Rapid identification of mycobacteria to species level by PCR-restriction fragment length polymorphism analysis of the hsp65 gene and proposition of an algorithm to differentiate 34 mycobacterial species. J. Clin. Microbiol. 35:29692973.
15. 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.
16. Evans, K. D.,, A. S. Nakasone,, P. A. Sutherland,, L. M. dela Maza,, and E. M. Peterson. 1992. Identification of Mycobacterium tuberculosisand Mycobacterium avium-M. intracellulare directly from primary BACTEC cultures by using acridinium- ester-labeled DNA probes. J. Clin. Microbiol. 30:24272431.
17. Gen-Probe, Inc. 1995. AccuProbe culture confirmation test for mycobacteria, package insert. Gen- Probe, Inc., San Diego, CA.
18. Goto, M.,, S. Oka,, K. Okuzumi,, S. Kimur,, and K. Shimada. 1991. Evaluation of acridinium ester- labeled DNA probes for identification of Mycobacterium tuberculosis and Mycobacterium avium-Mycobacterium intracellulare complex in culture. J. Clin. Microbiol. 29:24732476.
19. Musial, C. E.,, L. S. Tice,, L. Stockman,, and G. P. Roberts. 1988. Identification of mycobacteria from culture by using the Gen-Probe rapid diagnostic system for Mycobacterium avium complex and Mycobacterium tuberculosis complex. J. Clin. Microbiol. 26:21202123.
20. Richter, E.,, S. Niemann,, S. Rüsch-Gerdes,, and S. Hoffner. 1999. Identification of Mycobacterium kansasii by using a DNA probe (AccuProbe) and molecular techniques. J. Clin. Microbiol. 37:964970.
21. Alexander, B. D.,, E. Dodds-Ashley,, L. B. Reller,, and S. D. Reed. 2006. Cost savings with implementation of PNA FISH testing for identification of Candida albicans in blood cultures. Diagn. Microbiol. Infect. Dis. 54:277282.
22. Forrest, G. N.,, S. Mehta,, E. Weekes,, D. P. Lincalis,, J. K. Johnson,, and R. A. Venezia. 2006. Impact of rapid in situ hybridization testing on coagulase- negative staphylococci positive blood cultures. J. Antimicrob. Chemother. 58:143158.
23. Oliveira, K.,, S. M. Brecher,, A. Durbin,, D. S. Shapiro,, D. R. Schwartz,, P. C. De Girolami,, J. Dakos,, G. W. Procop,, D. Wilson,, C. S. Hanna,, G. Haase,, H. Peltroche-Llacsahuanga,, K. C. Chapin,, M. C. Musgnug,, M. H. Levi,, C. Shoemaker,, and H. Stender. 2003. Direct identification of Staphylococcus aureus from positive blood culture bottles. J. Clin. Microbiol. 41:889891.
24. Oliveira, K.,, G. W. Procop,, D. Wilson,, J. Coull,, and H. Stender. 2002. Rapid identification of Staphylococcus aureus directly from blood cultures by fluorescence in situ hybridization with peptide nucleic acid probes. J. Clin. Microbiol. 40:247251.
25. Rigby, S.,, G. W. Procop,, G. Haase,, D. Wilson,, G. Hall,, C. Kurtzman,, K. Oliveira,, S. Von Oy,, J. J. Hyldig-Nielsen,, J. Coull,, and H. Stender. 2002. Fluorescence in situ hybridization with peptide nucleic acid probes for rapid identification of Candida albicans directly from blood culture bottles. J. Clin. Microbiol. 40:21822186.
26. Sogarrd, M.,, D. S. Hansen,, M. J. Fiandaca,, H. Stender,, and H. C. Schonheyder. 2007. Peptide nucleic acid fluorescence in situ hybridization for rapid detection of Klebsiella pneumoniae from positive blood cultures. J. Med. Microbiol. 56:914917.
27. Wilson, D. A.,, M. J. Joyce,, L. S. Hall,, L. B. Reller,, G. D. Roberts,, G. S. Hall,, B. D. Alexander,, and G. W. Procop. 2005. Multicenter evaluation of a Candida albicans peptide nucleic acid in situ hybridization probe for characterization of yeast isolates from blood cultures. J. Clin. Microbiol. 43:29092921.

Tables

Generic image for table
Untitled

Citation: Garcia L. 2010. Molecular Methods for Identification of Cultured Microorganisms, p 366-379. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch12.3
Generic image for table
Untitled

Citation: Garcia L. 2010. Molecular Methods for Identification of Cultured Microorganisms, p 366-379. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch12.3
Generic image for table
Table 12.3.3-1

Reagents for AccuProbe culture confirmation

All reagents are stable for approximately 1 year from date of manufacture.

After pouch is opened, stable for 2 months.

Citation: Garcia L. 2010. Molecular Methods for Identification of Cultured Microorganisms, p 366-379. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch12.3
Generic image for table
Table 12.3.3-2

Control organisms for AccuProbe mycobacterial culture confirmation tests

Citation: Garcia L. 2010. Molecular Methods for Identification of Cultured Microorganisms, p 366-379. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch12.3

This is a required field
Please enter a valid email address
Please check the format of the address you have entered.
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error