ArticlePloS one2012
Detection of bacterial 16S rRNA and identification of four clinically important bacteria by real-time PCR.
Article in PloS one, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 106 papers.
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Who cites it
106 citing papers in PubMed.
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- Gut microbial composition varies with host metabolic phenotype in juvenile Atlantic salmon.The Journal of experimental biology · 2026Article
- CRISPR in MOF Formulation with Enhanced Stability, Activity, and Altered PAM Specificity for Broad-Spectrum Diagnosis of Bacterial Sepsis.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- The transcriptomic response of Staphylococcus equorum KS1030 to Lincomycin stress reveals transporters associated with horizontal gene transfer.Scientific reports · 2025Article
- Bsal susceptibility depends on host origin but not on skin microbiota in captive Pleurodeles waltl.Animal microbiome · 2025Article
- Utilization of clove and cinnamon essential oils as an alternative to inhibit MDR and biofilm producing E. coli from raw chicken meat.Scientific reports · 2025Article
- Blood Microbiome Analysis Reveals Biomarkers of Treatment Response in Drug-Naïve Patients with First-Episode Psychosis: A Pilot Study.Microorganisms · 2025Article
- Microbiome diversity of low biomass skin sites is captured by metagenomics but not 16S amplicon sequencing.bioRxiv : the preprint server for biology · 2025Article
- Correlation ofApplied and environmental microbiology · 2025Article
- New Insights into the Synergistic Interaction Between PseudomonasMicroorganisms · 2025Article
- Unveiling a hidden threat: whole genome sequencing exposes the cross-hospital spread of extensively drug-resistantFrontiers in microbiology · 2025Article
- OralFrontiers in dental medicine · 2025Article
- Assessment of absolute abundance in mother-infant gut microbiome using marine-sourced bacterial DNA spike-in and comparison with conventional quantification methods.Microbiome research reports · 2025Article
- Antimicrobial Sub-MIC induces Staphylococcus aureus biofilm formation without affecting the bacterial count.BMC infectious diseases · 2024Article
- Article
- Bacterial community of kimchi added with seafood based on culture-dependent investigations.Heliyon · 2024Article
46 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
9 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Within the paradigm of clinical infectious disease research, Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa represent the four most clinically relevant, and hence most extensively studied bacteria. Current culture-based methods for identifying these organisms are slow and cumbersome, and there is increasing need for more rapid and accurate molecular detection methods. Using bioinformatic tools, 962,279 bacterial 16S rRNA gene sequences were aligned, and regions of homology were selected to generate a set of real-time PCR primers that target 93.6% of all bacterial 16S rRNA sequences published to date. A set of four species-specific real-time PCR primer pairs were also designed, capable of detecting less than 100 genome copies of A. baumannii, E. coli, K. pneumoniae, and P. aeruginosa. All primers were tested for specificity in vitro against 50 species of Gram-positive and -negative bacteria. Additionally, the species-specific primers were tested against a panel of 200 clinical isolates of each species, randomly selected from a large repository of clinical isolates from diverse areas and sources. A comparison of culture and real-time PCR demonstrated 100% concordance. The primers were incorporated into a rapid assay capable of positive identification from plate or broth cultures in less than 90 minutes. Furthermore, our data demonstrate that current targets, such as the uidA gene in E.coli, are not suitable as species-specific genes due to sequence variation. The assay described herein is rapid, cost-effective and accurate, and can be easily incorporated into any research laboratory capable of real-time PCR.
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