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Related Articles Utility of Pyrosequencing in Identifying Bacteria Directly from Positive Blood Culture Bottles.
J Clin Microbiol. 2008 Dec 17;
Authors: Jordan JA, Jones-Laughner J, Durso MB
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 sub-culturing, would allow the use of more directed empiric therapy and thus reduce the patient's exposure to unnecessary or ineffective antibiotics sooner. The study had two objectives: to identify new targets to improve our ability to differentiate among certain enteric gram-negative rods or among certain Streptococcus species, and to determine whether real-time PCR and pyrosequencing could accurately identify organisms directly from positive blood culture bottles compared to 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 pyrosequencing and culture-based identifications were concordant for 264/270 (97.8%) bacteria with 3 failed sequences, and 3 sequences without match. Additionally, compared to the universal 16S rRNA gene targets, the new 23S rRNA gene targets greatly improved our ability to differentiate among 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 empiric therapy in patients and would promote responsible antibiotic stewardship.
PMID: 19091813 [PubMed - as supplied by publisher]
More...
Related Articles Utility of Pyrosequencing in Identifying Bacteria Directly from Positive Blood Culture Bottles.
J Clin Microbiol. 2008 Dec 17;
Authors: Jordan JA, Jones-Laughner J, Durso MB
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 sub-culturing, would allow the use of more directed empiric therapy and thus reduce the patient's exposure to unnecessary or ineffective antibiotics sooner. The study had two objectives: to identify new targets to improve our ability to differentiate among certain enteric gram-negative rods or among certain Streptococcus species, and to determine whether real-time PCR and pyrosequencing could accurately identify organisms directly from positive blood culture bottles compared to 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 pyrosequencing and culture-based identifications were concordant for 264/270 (97.8%) bacteria with 3 failed sequences, and 3 sequences without match. Additionally, compared to the universal 16S rRNA gene targets, the new 23S rRNA gene targets greatly improved our ability to differentiate among 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 empiric therapy in patients and would promote responsible antibiotic stewardship.
PMID: 19091813 [PubMed - as supplied by publisher]
More...