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Title: Use of 16S rRNA gene for identification of a broad range of clinically relevant bacterial pathogens

Journal Article · · PLoS ONE
 [1];  [2];  [3];  [4];  [5];  [6];  [6];  [2];  [1];  [7]
  1. Univ. of California San Francisco, San Francisco, CA (United States). Dept. of Medicine
  2. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Earth Sciences Div.
  3. Univ. of California, Berkeley, CA (United States)
  4. Victoria Univ. of Wellington, Wellington (New Zealand). School of BIological Sciences
  5. San Francisco General Hospital, San Francisco, CA (United States). Dept. of Medicine
  6. Univ. of California San Francisco, San Francisco, CA (United States). Clinical Microbiology Lab.
  7. Charité, Campus Benjamin Franklin, Berlin (Germany)

According to World Health Organization statistics of 2011, infectious diseases remain in the top five causes of mortality worldwide. However, despite sophisticated research tools for microbial detection, rapid and accurate molecular diagnostics for identification of infection in humans have not been extensively adopted. Time-consuming culture-based methods remain to the forefront of clinical microbial detection. The 16S rRNA gene, a molecular marker for identification of bacterial species, is ubiquitous to members of this domain and, thanks to ever-expanding databases of sequence information, a useful tool for bacterial identification. In this study, we assembled an extensive repository of clinical isolates (n = 617), representing 30 medically important pathogenic species and originally identified using traditional culture-based or non-16S molecular methods. This strain repository was used to systematically evaluate the ability of 16S rRNA for species level identification. To enable the most accurate species level classification based on the paucity of sequence data accumulated in public databases, we built a Naïve Bayes classifier representing a diverse set of high-quality sequences from medically important bacterial organisms. We show that for species identification, a model-based approach is superior to an alignment based method. Overall, between 16S gene based and clinical identities, our study shows a genus-level concordance rate of 96% and a species-level concordance rate of 87.5%. We point to multiple cases of probable clinical misidentification with traditional culture based identification across a wide range of gram-negative rods and gram-positive cocci as well as common gram-negative cocci.

Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
USDOE; National Institute of Health
Grant/Contract Number:
AC02-05CH11231
OSTI ID:
1212477
Journal Information:
PLoS ONE, Journal Name: PLoS ONE Journal Issue: 2 Vol. 10; ISSN 1932-6203
Publisher:
Public Library of ScienceCopyright Statement
Country of Publication:
United States
Language:
English

References (114)

Evaluation of the ability of a commercial system to identifyAcinetobacter genomic species journal April 1996
Comparative evaluation of microbial profiles of oral samples obtained at different collection time points and using different methods journal September 2020
Evaluation of 16SpathDB 2.0, an automated 16S rRNA gene sequence database, using 689 complete bacterial genomes journal February 2014
Investigating a common approach to DNA profile interpretation using probabilistic software journal May 2015
The rpoB gene as a tool for clinical microbiologists journal January 2009
Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms journal March 2012
Structure, function and diversity of the healthy human microbiome journal June 2012
Evaluation of a biochemical test scheme for identifying clinical isolates of Enterococcus faecalis and Enterococcus faecium journal November 2001
The Epidemiology of Sepsis in the United States from 1979 through 2000 journal April 2003
RIDOM: Ribosomal Differentiation of Medical Micro-organisms Database journal January 2002
The Ribosomal Database Project: improved alignments and new tools for rRNA analysis journal January 2009
Update on the Burkholderia cepacia complex journal January 2005
Measuring the effect of inappropriate initial antibiotic therapy on outcomes of patients with Gram-negative sepsis: An imprecise science* journal January 2011
Towards a Phylogeny and Definition of Species at the Molecular Level within the Genus Mycobacterium journal October 1990
Serratia marcescens journal November 1997
Evaluation of three phenotypic identification systems for clinical isolates of Raoultella ornithinolytica journal January 2011
Effect of a very low-energy diet on moderate and severe obstructive sleep apnoea: case reports journal February 2011
Naive Bayesian Classifier for Rapid Assignment of rRNA Sequences into the New Bacterial Taxonomy journal June 2007
Greengenes, a Chimera-Checked 16S rRNA Gene Database and Workbench Compatible with ARB journal July 2006
Molecular Epidemiology of Tuberculosis: Current Insights journal October 2006
What Happened to the Streptococci: Overview of Taxonomic and Nomenclature Changes journal October 2002
Impact of 16S rRNA Gene Sequence Analysis for Identification of Bacteria on Clinical Microbiology and Infectious Diseases journal October 2004
Natural competence in the genus Streptococcus: evidence that streptococci can change pherotype by interspecies recombinational exchanges. journal November 1997
Using Multilocus Sequence Data To Define the Pneumococcus journal August 2005
Application of SmartGene IDNS Software to Partial 16S rRNA Gene Sequences for a Diverse Group of Bacteria in a Clinical Laboratory journal October 2006
16S Ribosomal DNA Sequence Analysis of a Large Collection of Environmental and Clinical Unidentifiable Bacterial Isolates journal January 2000
Evaluation of the VITEK 2 System for the Identification and Susceptibility Testing of Three Species of Nonfermenting Gram-Negative Rods Frequently Isolated from Clinical Samples journal September 2001
Comparative Assessment of Genotyping Methods for Epidemiologic Study of Burkholderia cepacia Genomovar III journal September 2002
Evaluation of the MicroSeq System for Identification of Mycobacteria by 16S Ribosomal DNA Sequencing and Its Integration into a Routine Clinical Mycobacteriology Laboratory journal April 2003
Clinical Impact of a PCR Assay for Identification of Staphylococcus aureus and Determination of Methicillin Resistance Directly from Blood Cultures journal August 2003
Ribosomal DNA Sequencing for Identification of Aerobic Gram-Positive Rods in the Clinical Laboratory (an 18-Month Evaluation) journal September 2003
PCR-Based Assay for Differentiation of Pseudomonas aeruginosa from Other Pseudomonas Species Recovered from Cystic Fibrosis Patients journal May 2004
Use of the MicroSeq 500 16S rRNA Gene-Based Sequencing for Identification of Bacterial Isolates That Commercial Automated Systems Failed To Identify Correctly journal February 2005
Development of a Multilocus Sequence Typing Scheme for Characterization of Clinical Isolates of Acinetobacter baumannii journal September 2005
Gemella bacteraemia characterised by 16S ribosomal RNA gene sequencing journal September 2003
Estimating Health Care-Associated Infections and Deaths in U.S. Hospitals, 2002 journal March 2007
The effect of training set on the classification of honey bee gut microbiota using the Naïve Bayesian Classifier journal January 2012
The Effects of Alignment Quality, Distance Calculation Method, Sequence Filtering, and Region on the Analysis of 16S rRNA Gene-Based Studies journal July 2010
Exploring Microbial Diversity and Taxonomy Using SSU rRNA Hypervariable Tag Sequencing journal November 2008
Evaluation of 16S rDNA-Based Community Profiling for Human Microbiome Research journal June 2012
Clinical Importance of Delays in the Initiation of Appropriate Antibiotic Treatment for Ventilator-Associated Pneumonia journal July 2002
Laboratory Maintenance of Bordetella pertussis journal November 2009
Evaluation of the ability of a commercial system to identifyAcinetobacter genomic species journal April 1996
A multiplex real-time PCR assay for rapid detection and differentiation of 25 bacterial and fungal pathogens from whole blood samples journal November 2007
Rapid diagnosis of Clostridium difficile infection by multiplex real-time PCR journal April 2011
Molecular Typing of Bacteria Directly from Cerebrospinal Fluid journal September 2000
Serratia marcescens journal October 1987
A Rapid PCR Based Method to Distinguish between Lactococcus and Enterococcus journal December 2000
Evaluation of 16SpathDB 2.0, an automated 16S rRNA gene sequence database, using 689 complete bacterial genomes journal February 2014
16S rRNA sequencing in routine bacterial identification: A 30-month experiment journal December 2006
The rpoB gene as a tool for clinical microbiologists journal January 2009
Impact of training sets on classification of high-throughput bacterial 16s rRNA gene surveys journal June 2011
Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms journal March 2012
Structure, function and diversity of the healthy human microbiome journal June 2012
Microbial Dysbiosis During Simian Immunodeficiency Virus Infection is Partially Reverted with Combination Anti-retroviral Therapy journal April 2020
Evaluation of a biochemical test scheme for identifying clinical isolates of Enterococcus faecalis and Enterococcus faecium journal November 2001
The rate and cost of hospital-acquired infections occurring in patients admitted to selected specialties of a district general hospital in England and the national burden imposed journal March 2001
The Epidemiology of Sepsis in the United States from 1979 through 2000 journal April 2003
Serratia marcescens journal August 1979
The growing burden of antimicrobial resistance journal September 2008
RIDOM: Ribosomal Differentiation of Medical Micro-organisms Database journal January 2002
SILVA: a comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB journal November 2007
The Ribosomal Database Project: improved alignments and new tools for rRNA analysis journal January 2009
Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies journal August 2012
The epidemiology of sepsis in the United States from 1979 through 2000* journal August 2003
Update on the Burkholderia cepacia complex journal January 2005
Inappropriate antibiotic therapy in Gram-negative sepsis increases hospital length of stay* journal January 2011
Towards a Phylogeny and Definition of Species at the Molecular Level within the Genus Mycobacterium journal October 1990
Serratia marcescens journal November 1997
Evaluation of three phenotypic identification systems for clinical isolates of Raoultella ornithinolytica journal January 2011
Neisseria meningitidis PorA variable regions: rapid detection of P1·7 and P1·19 variants by PCR : Rapid detection of journal July 2007
Bacterial Community Variation in Human Body Habitats Across Space and Time journal November 2009
Naive Bayesian Classifier for Rapid Assignment of rRNA Sequences into the New Bacterial Taxonomy journal June 2007
Introducing mothur: Open-Source, Platform-Independent, Community-Supported Software for Describing and Comparing Microbial Communities journal October 2009
Diversity of 16S rRNA Genes within Individual Prokaryotic Genomes journal April 2010
Greengenes, a Chimera-Checked 16S rRNA Gene Database and Workbench Compatible with ARB journal July 2006
Molecular Epidemiology of Tuberculosis: Current Insights journal October 2006
Endocarditis Due to Rare and Fastidious Bacteria journal January 2001
What Happened to the Streptococci: Overview of Taxonomic and Nomenclature Changes journal October 2002
Impact of 16S rRNA Gene Sequence Analysis for Identification of Bacteria on Clinical Microbiology and Infectious Diseases journal October 2004
Genetic Relationships between Clinical Isolates of Streptococcus pneumoniae, Streptococcus oralis, and Streptococcus mitis: Characterization of "Atypical" Pneumococci and Organisms Allied to S. mitis Harboring S. pneumoniae Virulence Factor-Encoding Genes journal March 2000
Using Multilocus Sequence Data To Define the Pneumococcus journal August 2005
Application of SmartGene IDNS Software to Partial 16S rRNA Gene Sequences for a Diverse Group of Bacteria in a Clinical Laboratory journal October 2006
Automated Identification of Medically Important Bacteria by 16S rRNA Gene Sequencing Using a Novel Comprehensive Database, 16SpathDB journal March 2011
16S Ribosomal DNA Sequence Analysis of a Large Collection of Environmental and Clinical Unidentifiable Bacterial Isolates journal January 2000
Comparison of Three Commercial Rapid Identification Systems for the Unusual Gram-Positive Cocci Dolosigranulum pigrum, Ignavigranum ruoffiae, andFacklamia Species journal January 2000
Evaluation of the VITEK 2 System for the Identification and Susceptibility Testing of Three Species of Nonfermenting Gram-Negative Rods Frequently Isolated from Clinical Samples journal September 2001
Comparative Assessment of Genotyping Methods for Epidemiologic Study of Burkholderia cepacia Genomovar III journal September 2002
Evaluation of the MicroSeq System for Identification of Mycobacteria by 16S Ribosomal DNA Sequencing and Its Integration into a Routine Clinical Mycobacteriology Laboratory journal April 2003
Usefulness of the MicroSeq 500 16S Ribosomal DNA-Based Bacterial Identification System for Identification of Clinically Significant Bacterial Isolates with Ambiguous Biochemical Profiles journal May 2003
Clinical Impact of a PCR Assay for Identification of Staphylococcus aureus and Determination of Methicillin Resistance Directly from Blood Cultures journal August 2003
Ribosomal DNA Sequencing for Identification of Aerobic Gram-Positive Rods in the Clinical Laboratory (an 18-Month Evaluation) journal September 2003
Confirmation of Nontypeable Streptococcus pneumoniae-Like Organisms Isolated from Outbreaks of Epidemic Conjunctivitis as Streptococcus pneumoniae journal September 2003
PCR-Based Assay for Differentiation of Pseudomonas aeruginosa from Other Pseudomonas Species Recovered from Cystic Fibrosis Patients journal May 2004
Use of the MicroSeq 500 16S rRNA Gene-Based Sequencing for Identification of Bacterial Isolates That Commercial Automated Systems Failed To Identify Correctly journal February 2005
Development of a Multilocus Sequence Typing Scheme for Characterization of Clinical Isolates of Acinetobacter baumannii journal September 2005
Diversity of 16S rRNA Genes within Individual Prokaryotic Genomes journal August 2010
Natural competence in the genus Streptococcus: evidence that streptococci can change pherotype by interspecies recombinational exchanges. journal November 1997
Development of a Multilocus Sequence Typing Scheme for Characterization of Clinical Isolates of Acinetobacter baumannii journal June 2007
Gemella bacteraemia characterised by 16S ribosomal RNA gene sequencing journal September 2003
Prospects for the Future Using Genomics and Proteomics in Clinical Microbiology journal October 2011
Community Factors in the Development of Antibiotic Resistance journal April 2007
Inappropriate Antibiotic Therapy In Gram-Negative Sepsis Increases Hospital Length Of Stay conference May 2010
Estimating Health Care-Associated Infections and Deaths in U.S. Hospitals, 2002 journal March 2007
Species-level classification of the vaginal microbiome journal December 2012
The effect of training set on the classification of honey bee gut microbiota using the Naïve Bayesian Classifier journal January 2012
Identification of pathogenic Leptospira species by conventional or real-time PCR and sequencing of the DNA gyrase subunit B encoding gene journal October 2006
The Effects of Alignment Quality, Distance Calculation Method, Sequence Filtering, and Region on the Analysis of 16S rRNA Gene-Based Studies journal July 2010
Exploring Microbial Diversity and Taxonomy Using SSU rRNA Hypervariable Tag Sequencing journal November 2008
Evaluation of 16S rDNA-Based Community Profiling for Human Microbiome Research journal June 2012
Species-Level Analysis of DNA Sequence Data from the NIH Human Microbiome Project journal October 2012
Clinical Importance of Delays in the Initiation of Appropriate Antibiotic Treatment for Ventilator-Associated Pneumonia journal July 2002
Impact of Rapid Methicillin-Resistant Staphylococcus aureus Polymerase Chain Reaction Testing on Mortality and Cost Effectiveness in Hospitalized Patients with Bacteraemia: A Decision Model journal July 2010
Влияние мутаций по внеклеточной нуклеазе на свойства пигментообразующего и беспигментного штаммов Serratia marcescens journal January 2016

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