Extended Bibliography: |
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Ref #: |
95508 |
Author(s): |
Anthony,R.M.;Brown,T.J.;French,G.L. |
Journal: |
J Clin Microbiol |
Title: |
Rapid diagnosis of bacteremia by universal amplification of 23S ribosomal DNA followed by hybridization to an oligonucleotide array |
Volume: |
38 |
Page(s): |
781-8 |
Year: |
2000 |
Keyword(s): |
GENBANK/AF146762
GENBANK/AF146763
GENBANK/AF146764
GENBANK/AF146765
GENBANK/AF146766
GENBANK/AF146768
GENBANK/AF146769
GENBANK/AF146770
GENBANK/AF146771
GENBANK/AF146772
GENBANK/AF146773
GENBANK/AF146774
Bacteremia/*diagnosis/microbiology
Bacteria/classification/*genetics/isolation & purification
DNA Primers
DNA, Bacterial/analysis
DNA, Ribosomal/analysis
Humans
Molecular Sequence Data
*Oligonucleotide Array Sequence Analysis
Polymerase Chain Reaction/*methods
Predictive Value of Tests
RNA, Ribosomal, 23S/*genetics
|
Remarks: |
The rapid identification of bacteria in blood cultures and other clinical specimens is important for patient management and antimicrobial therapy. We describe a rapid (<4 h) detection and identification system that uses universal PCR primers to amplify a variable region of bacterial 23S ribosomal DNA, followed by reverse hybridization of the products to a panel of oligonucleotides. This procedure was successful in discriminating a range of bacteria in pure cultures. When this procedure was applied directly to 158 unselected positive blood culture broths on the day when growth was detected, 125 (79.7%) were correctly identified, including 4 with mixed cultures. Nine (7.2%) yielded bacteria for which no oligonucleotide targets were present in the oligonucleotide panel, and 16 culture-positive broths (10.3%) produced no PCR product. In seven of the remaining eight broths, streptococci were identified but not subsequently grown, and one isolate of Staphylococcus aureus was misidentified as a coagulase-negative staphylococcus. The accuracy, range, and discriminatory power of the assay can be continually extended by adding further oligonucleotides to the panel without significantly increasing complexity or cost. |
URL: |
10655385 |
|
Ref #: |
81199 |
Author(s): |
Becker,K.;Harmsen,D.;Mellmann,A.;Meier,C.;Schumann,P.;Peters,G.;von Eiff,C. |
Journal: |
J Clin Microbiol |
Title: |
Development and evaluation of a quality-controlled ribosomal sequence database for 16S ribosomal DNA-based identification of Staphylococcus species |
Volume: |
42 |
Page(s): |
4988-95 |
Year: |
2004 |
Keyword(s): |
GENBANK/AY688029
GENBANK/AY688030
GENBANK/AY688031
GENBANK/AY688032
GENBANK/AY688033
GENBANK/AY688034
GENBANK/AY688035
GENBANK/AY688036
GENBANK/AY688037
GENBANK/AY688038
GENBANK/AY688039
GENBANK/AY688040
GENBANK/AY688041
GENBANK/AY688042
GENBANK/AY688043
GENBANK/AY688044
GENBANK/AY688045
GENBANK/AY688046
GENBANK/AY688047
GENBANK/AY688048
GENBANK/AY688049
GENBANK/AY688050
GENBANK/AY688051
GENBANK/AY688052
GENBANK/AY688053
GENBANK/AY688054
GENBANK/AY688055
GENBANK/AY688056
GENBANK/AY688057
GENBANK/AY688058
GENBANK/AY688059
GENBANK/AY688060
GENBANK/AY688061
GENBANK/AY688062
GENBANK/AY688063
GENBANK/AY688064
GENBANK/AY688065
GENBANK/AY688066
GENBANK/AY688067
GENBANK/AY688068
GENBANK/AY688069
GENBANK/AY688070
GENBANK/AY688071
GENBANK/AY688072
GENBANK/AY688073
GENBANK/AY688074
GENBANK/AY688075
GENBANK/AY688076
GENBANK/AY688077
GENBANK/AY688078
GENBANK/AY688079
GENBANK/AY688080
GENBANK/AY688081
GENBANK/AY688082
GENBANK/AY688083
GENBANK/AY688084
GENBANK/AY688085
GENBANK/AY688086
GENBANK/AY688087
GENBANK/AY688088
GENBANK/AY688089
GENBANK/AY688090
GENBANK/AY688091
GENBANK/AY688092
GENBANK/AY688093
GENBANK/AY688094
GENBANK/AY688095
GENBANK/AY688096
GENBANK/AY688097
GENBANK/AY688098
GENBANK/AY688099
GENBANK/AY688100
GENBANK/AY688101
GENBANK/AY688102
GENBANK/AY688103
GENBANK/AY688104
GENBANK/AY688105
GENBANK/AY688106
GENBANK/AY688107
GENBANK/AY688108
GENBANK/AY688109
*Bacterial Typing Techniques
DNA, Bacterial/analysis
DNA, Ribosomal/analysis
*Databases, Nucleic Acid
Genes, rRNA
Humans
Molecular Sequence Data
RNA, Ribosomal, 16S/*genetics
Sequence Analysis, DNA
Species Specificity
Staphylococcal Infections/*microbiology
Staphylococcus/*classification/genetics/metabolism
|
Remarks: |
To establish an improved ribosomal gene sequence database as part of the Ribosomal Differentiation of Microorganisms (RIDOM) project and to overcome the drawbacks of phenotypic identification systems and publicly accessible sequence databases, both strands of the 5' end of the 16S ribosomal DNA (rDNA) of 81 type and reference strains comprising all validly described staphylococcal (sub)species were sequenced. Assuming a normal distribution for pairwise distances of all unique staphylococcal sequences and choosing a reporting criterion of > or =98.7% similarity for a "distinct species," a statistical error probability of 1.0% was calculated. To evaluate this database, a 16S rDNA fragment (corresponding to Escherichia coli positions 54 to 510) of 55 clinical Staphylococcus isolates (including those of the small-colony variant phenotype) were sequenced and analyzed by the RIDOM approach. Of these isolates, 54 (98.2%) had a similarity score above the proposed threshold using RIDOM; 48 (87.3%) of the sequences gave a perfect match, whereas 83.6% were found by searching National Center for Biotechnology Information (NCBI) database entries. In contrast to RIDOM, which showed four ambiguities at the species level (mainly concerning Staphylococcus intermedius versus Staphylococcus delphini), the NCBI database search yielded 18 taxon-related ambiguities and showed numerous matches exhibiting redundant or unspecified entries. Comparing molecular results with those of biochemical procedures, ID 32 Staph (bioMerieux, Marcy I'Etoile, France) and VITEK 2 (bioMerieux) failed to identify 13 (23.6%) and 19 (34.5%) isolates, respectively, due to incorrect identification and/or categorization below acceptable values. In contrast to phenotypic methods and the NCBI database, the novel high-quality RIDOM sequence database provides excellent identification of staphylococci, including rarely isolated species and phenotypic variants. |
URL: |
15528685 |
|
Ref #: |
13707 |
Author(s): |
Anthony,R.M.;Brown,T.J.;French,G.L. |
Journal: |
J Clin Microbiol |
Title: |
Rapid diagnosis of bacteremia by universal amplification of 23S ribosomal DNA followed by hybridization to an oligonucleotide array |
Volume: |
38 |
Page(s): |
781-8 |
Year: |
2000 |
Keyword(s): |
GENBANK/AF146762
GENBANK/AF146763
GENBANK/AF146764
GENBANK/AF146765
GENBANK/AF146766
GENBANK/AF146768
GENBANK/AF146769
GENBANK/AF146770
GENBANK/AF146771
GENBANK/AF146772
GENBANK/AF146773
GENBANK/AF146774
Bacteremia/*diagnosis/microbiology
Bacteria/classification/*genetics/isolation & purification
DNA Primers
DNA, Bacterial/analysis
DNA, Ribosomal/analysis
Human
Molecular Sequence Data
*Oligonucleotide Array Sequence Analysis
Polymerase Chain Reaction/*methods
Predictive Value of Tests
RNA, Ribosomal, 23S/*genetics
|
Remarks: |
The rapid identification of bacteria in blood cultures and other clinical specimens is important for patient management and antimicrobial therapy. We describe a rapid (<4 h) detection and identification system that uses universal PCR primers to amplify a variable region of bacterial 23S ribosomal DNA, followed by reverse hybridization of the products to a panel of oligonucleotides. This procedure was successful in discriminating a range of bacteria in pure cultures. When this procedure was applied directly to 158 unselected positive blood culture broths on the day when growth was detected, 125 (79.7%) were correctly identified, including 4 with mixed cultures. Nine (7.2%) yielded bacteria for which no oligonucleotide targets were present in the oligonucleotide panel, and 16 culture-positive broths (10.3%) produced no PCR product. In seven of the remaining eight broths, streptococci were identified but not subsequently grown, and one isolate of Staphylococcus aureus was misidentified as a coagulase-negative staphylococcus. The accuracy, range, and discriminatory power of the assay can be continually extended by adding further oligonucleotides to the panel without significantly increasing complexity or cost. |
URL: |
20122462 |
|
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