This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Jordan, J. A.
Right arrow Articles by Durso, M. B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Jordan, J. A.
Right arrow Articles by Durso, M. B.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, February 2009, p. 368-372, Vol. 47, No. 2
0095-1137/09/$08.00+0     doi:10.1128/JCM.01991-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Utility of Pyrosequencing in Identifying Bacteria Directly from Positive Blood Culture Bottles{triangledown}

J. A. Jordan,1,2* J. Jones-Laughner,2 and M. B. Durso2

University of Pittsburgh, School of Medicine, Department of Pathology,1 Magee-Women's Research Institute and Foundation, Pittsburgh, Pennsylvania2

Received 15 October 2008/ Returned for modification 29 November 2008/ Accepted 8 December 2008

Growth in liquid media is the gold standard for detecting microorganisms associated with bloodstream infections. The Gram stain provides the first clue as to the etiology of infection, with phenotypic identification completed 1 or 2 days later. Providing more detailed information than the Gram stain can impart, and in less time than subculturing, would allow the use of more directed empirical therapy and, thus, reduce the patient's exposure to unnecessary or ineffective antibiotics sooner. The study had two objectives, as follows: (i) to identify new targets to improve our ability to differentiate among certain enteric gram-negative rods or among certain Streptococcus species and (ii) to determine whether real-time PCR and pyrosequencing could as accurately identify organisms directly from positive blood culture bottles as culture-based methods. Two hundred and fifty-five consecutive positive blood culture bottles were included. The results showed a high level of agreement between the two approaches; of the 270 bacteria isolated from the 255 blood culture bottles, results for pyrosequencing and culture-based identifications were concordant for 264/270 (97.8%) bacteria with three failed sequences, and three sequences without match. Additionally, compared to the universal 16S rRNA gene target, the new 23S rRNA gene targets greatly improved our ability to differentiate among certain enteric gram-negative rods or among certain Streptococcus species. In conclusion, combining real-time PCR and pyrosequencing provided valuable information beyond that derived from the initial Gram stain and in less time than phenotypic culture-based identification. This strategy, if implemented, could result in a more directed empirical therapy in patients and would promote responsible antibiotic stewardship.

* Corresponding author. Present address: Department of Epidemiology and Biostatistics, School of Public Health and Health Services, George Washington University, 231 Ross Hall, 2300 I Street, NW, Washington, DC 20037. Phone: (202) 994-7062. Fax: (202) 994-0082. E-mail: sphjaj{at}gwumc.edu

{triangledown} Published ahead of print on 17 December 2008.

Journal of Clinical Microbiology, February 2009, p. 368-372, Vol. 47, No. 2
0095-1137/09/$08.00+0     doi:10.1128/JCM.01991-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»