ArticleApplied and environmental microbiology2007
Sensitive quantitative detection of commensal bacteria by rRNA-targeted reverse transcription-PCR.
Article in Applied and environmental microbiology, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 124 papers.
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Who cites it
124 citing papers in PubMed, 296 citations in OpenAlex.
- Trial
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- Efficacy of Bifidobacterium breve Fermented Milk in Maintaining Remission of Ulcerative Colitis.Digestive diseases and sciences · 2018Trial
- Probiotic reduces bacterial translocation in type 2 diabetes mellitus: A randomised controlled study.Scientific reports · 2017Trial
- Efficacy of perioperative synbiotics treatment for the prevention of surgical site infection after laparoscopic colorectal surgery: a randomized controlled trial.Surgery today · 2016Trial
- Influence of a probiotic mixture on antibiotic induced microbiota disturbances.World journal of gastroenterology · 2014Trial
- Significant changes in the intestinal environment after surgery in patients with colorectal cancer.Journal of gastrointestinal surgery : official journal of the Society for Surgery of the Alimentary Tract · 2013Trial
- Synbiotic therapy reduces the pathological gram-negative rods caused by an increased acetic acid concentration in the gut.Digestive diseases and sciences · 2012Trial
- Probiotics Exert Colonization Resistance AgainstMicroorganisms · 2026Article
- A reverse transcription-quantitative polymerase chain reaction system for evaluating intestinal butyrate production by fecal bacteria.Applied and environmental microbiology · 2026Article
- Developing a Novel Fermented Milk with Anti-Aging and Anti-Oxidative Properties UsingNutrients · 2025Article
- Characteristics of the gut microbiota in patients with advanced non-small cell lung cancer who responded to immune checkpoint inhibitors.Scientific reports · 2025Article
- Article
- A novel ionic liquid-based approach for DNA and RNA extraction simplifies sample preparation for bacterial diagnostics.Analytical and bioanalytical chemistry · 2024Article
- Molecular characterisation of virulence genes in bacterial pathogens from daycare centres in Ile-Ife, Nigeria: implications for infection control.BMC infectious diseases · 2024Article
- Duplex Reverse-Transcription Real-Time Polymerase Chain Reaction Assay Targeting 23S rRNA Single Nucleotide Polymorphisms for the Detection of Flea-Borne Rickettsioses.The American journal of tropical medicine and hygiene · 2024Article
- Characterization of faecal microbiota and serum inflammatory markers in dogs diagnosed with chronic enteropathy or small-cell lymphoma: a pilot study.Scientific reports · 2024Article
- Investigating the Efficacy of Kidney-ProtectiveAnimals : an open access journal from MDPI · 2024Article
64 more citing papers are in PubMed but not listed here.
Corrections and comments
- Erratum issued
Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A sensitive rRNA-targeted reverse transcription-quantitative PCR (RT-qPCR) method was developed for exact and sensitive enumeration of subdominant bacterial populations. Using group- or species-specific primers for 16S or 23S rRNA, analytical curves were constructed for Escherichia coli, Enterococcus faecalis, Staphylococcus aureus, Clostridium perfringens, and Pseudomonas aeruginosa, and the threshold cycle value was found to be linear up to an RNA amount of 10(-3) cell per RT-PCR. The number of bacteria in culture was determined by RT-qPCR, and the results correlated well with the CFU count over the range from 10(0) to 10(5) CFU. The bacterial counts obtained by RT-qPCR were the same as the CFU counts irrespective of the growth phase in vitro, except for C. perfringens during starvation periods; the viable cell counts obtained by using a combination of 4',6-diamidino-2-phenylindole (DAPI) staining and SYTO9-propidium iodide double staining were in good agreement with the RT-qPCR counts rather than with the CFU counts. The RT-qPCR method could detect endogenous Enterobacteriaceae and P. aeruginosa in feces of hospitalized patients (n = 38) at a level of 10(3) cells per g of feces, and for enumeration of S. aureus or P. aeruginosa spiked into human peripheral blood, the lower detection limit for RT-qPCR quantification of the bacteria was 2 cells per ml of blood, suggesting that this method was equivalent to the conventional culture method. As only 5 h was needed for RT-qPCR quantification, we suggest that rRNA-targeted RT-qPCR assays provide a sensitive and convenient system for quantification of commensal bacteria and for examining their possible invasion of a host.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.