ArticleBioMed research international2013
Development of a broad-range 23S rDNA real-time PCR assay for the detection and quantification of pathogenic bacteria in human whole blood and plasma specimens.
Article in BioMed research international, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 27 citations in OpenAlex.
- Preparation and pharmacokinetic evaluation of Staphylococcus phage COP-80B for treatment of periprosthetic joint infections in a mouse model.Virus research · 2025Article
- Establishment of a 23S rRNA assay for Brucella and its application in evaluating bacterial growth status.Veterinary research communications · 2025Article
- Loop-Mediated Isothermal Amplification of DNA (LAMP) as an Alternative Method for Determining Bacteria in Wound Infections.International journal of molecular sciences · 2023Review
- Nusbiarylins Inhibit Transcription and Target Virulence Factors in Bacterial PathogenInternational journal of molecular sciences · 2020Article
- Intestinal permeability and bacterial translocation in patients with liver disease, focusing on alcoholic aetiology: methods of assessment and therapeutic intervention.Therapeutic advances in gastroenterology · 2020Review
- No effects without causes: the Iron Dysregulation and Dormant Microbes hypothesis for chronic, inflammatory diseases.Biological reviews of the Cambridge Philosophical Society · 2018Review
- A Dormant Microbial Component in the Development of Preeclampsia.Frontiers in medicine · 2016Review
- Limit of detection of genomic DNA by conventional PCR for estimating the load of Staphylococcus aureus and Escherichia coli associated with bovine mastitis.Folia microbiologica · 2015Article
- The dormant blood microbiome in chronic, inflammatory diseases.FEMS microbiology reviews · 2015Review
- Individuality, phenotypic differentiation, dormancy and 'persistence' in culturable bacterial systems: commonalities shared by environmental, laboratory, and clinical microbiology.F1000Research · 2015Review
- Oral bacterial DNA findings in pericardial fluid.Journal of oral microbiology · 2014Article
- Culture-Negative Peritonitis and the Latest Diagnostic Techniques.Kidney diseases (Basel, Switzerland)Review
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Authors and funding
10 authors at 4 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Molecular methods are important tools in the diagnosis of bloodstream bacterial infections, in particular in patients treated with antimicrobial therapy, due to their quick turn-around time. Here we describe a new broad-range real-time PCR targeting the 23S rDNA gene and capable to detect as low as 10 plasmid copies per reaction of targeted bacterial 23S rDNA gene. Two commercially available DNA extraction kits were evaluated to assess their efficiency for the extraction of plasma and whole blood samples spiked with different amount of either Staphylococcus aureus or Escherichia coli, in order to find the optimal extraction method to be used. Manual QIAmp extraction method with enzyme pre-treatment resulted the most sensitive for detection of bacterial load. Sensitivity of this novel assay ranged between 10 and 10(3) CFU per PCR reaction for E. coli and S. aureus in human whole blood samples depending on the extraction methods used. Analysis of plasma samples showed a 10- to 100-fold reduction of bacterial 23S rDNA in comparison to the corresponding whole blood specimens, thus indicating that whole blood is the preferential sample type to be used in this real-time PCR protocol. Our results thus show that the 23S rDNA gene represents an optimal target for bacteria quantification in human whole blood.
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