ArticleJournal of clinical microbiology2008
Method To Detect Only Live Bacteria during PCR Amplification.
Article in Journal of clinical microbiology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
25 citing papers in PubMed, 101 citations in OpenAlex.
- Revisiting Strategies for Bacterial Enumeration: The Case for Viable but Nonculturable (VBNC) Cells in Supplement Products.Probiotics and antimicrobial proteins · 2026Review
- Integrating FRET and Molecular Dynamics Simulation for Single-Molecule Aptameric Detection of Staphylococcus aureus IsdA Surface Protein.Biotechnology journal · 2025Article
- An Integrated PMA Pretreatment Instrument for Simultaneous Quantitative Detection ofFoods (Basel, Switzerland) · 2025Article
- Illumina Sequencing in Conjunction with Propidium Monoazide to Identify Live Bacteria After Antiseptic Treatment in a Complex Oral Biofilm: A Study Using an Ex Vivo Supragingival Biofilm Model.Antibiotics (Basel, Switzerland) · 2024Article
- Article
- PCR Mediated Nucleic Acid Molecular Recognition Technology for Detection of Viable and Dead Foodborne Pathogens.Foods (Basel, Switzerland) · 2022Article
- Review
- Article
- A Propidium Monoazide (PMAxx)-Droplet Digital PCR (ddPCR) for the Detection of ViableMicroorganisms · 2022Article
- Detection ofFrontiers in microbiology · 2022Article
- Electrochromogenic Detection of Live Bacteria Using Soluble and Insoluble Prussian Blue.ACS omega · 2021Article
- Combining Culture-Dependent and Culture-Independent Methods: New Methodology Insight on theFoods (Basel, Switzerland) · 2021Article
- Genome-Wide fitness analysis of group B Streptococcus in human amniotic fluid reveals a transcription factor that controls multiple virulence traits.PLoS pathogens · 2021Article
- A Novel, Rapid, and Simple PMA-qPCR Method for Detection and Counting of ViableJournal of veterinary research · 2020Article
- Spiking a Silty-Sand Reference Soil with Bacterial DNA: Limits and Pitfalls in the Discrimination of Live and Dead Cells When Applying Ethidium Monoazide (EMA) Treatment.Current microbiology · 2019Article
- Viable and Total Bacterial Populations Undergo Equipment- and Time-Dependent Shifts during Milk Processing.Applied and environmental microbiology · 2019Article
- Schrödinger's microbes: Tools for distinguishing the living from the dead in microbial ecosystems.Microbiome · 2017Review
- Innovative Use of Palladium Compounds To Selectively Detect Live Enterobacteriaceae in Milk by PCR.Applied and environmental microbiology · 2016Article
- Distribution and Diversity of Pathogenic Leptospira Species in Peri-domestic Surface Waters from South Central Chile.PLoS neglected tropical diseases · 2016Article
- A novel mechanism for direct real-time polymerase chain reaction that does not require DNA isolation from prokaryotic cells.Scientific reports · 2016Article
Corrections and comments
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Ethidium monoazide (EMA) is a DNA cross-linking agent and eukaryotic topoisomerase II poison. We previously reported that the treatment of EMA with visible light irradiation (EMA + Light) directly cleaved chromosomal DNA of Escherichia coli (T. Soejima, K. Iida, T. Qin, H. Taniai, M. Seki, A. Takade, and S. Yoshida, Microbiol. Immunol. 51:763-775, 2007). Herein, we report that EMA + Light randomly cleaved chromosomal DNA of heat-treated, but not live, Listeria monocytogenes cells within 10 min of treatment. When PCR amplified DNA that was 894 bp in size, PCR final products from 10(8) heat-treated L. monocytogenes were completely suppressed by EMA + Light. When target DNA was short (113 bp), like the hly gene of L. monocytogenes, DNA amplification was not completely suppressed by EMA + Light only. Thus, we used DNA gyrase/topoisomerase IV and mammalian topoisomerase poisons (here abbreviated as T-poisons) together with EMA + Light. T-poisons could penetrate heat-treated, but not live, L. monocytogenes cells within 30 min to cleave chromosomal DNA by poisoning activity. The PCR product of the hly gene from 10(8) heat-treated L. monocytogenes cells was inhibited by a combination of EMA + Light and T-poisons (EMA + Light + T-poisons), but those from live bacteria were not suppressed. As a model for clinical application to bacteremia, we tried to discriminate live and antibiotic-treated L. monocytogenes cells present in human blood. EMA + Light + T-poisons completely suppressed the PCR product from 10(3) to 10(7) antibiotic-treated L. monocytogenes cells but could detect 10(2) live bacteria. Considering the prevention and control of food poisoning, this method was applied to discriminate live and heat-treated L. monocytogenes cells spiked into pasteurized milk. EMA + Light + T-poisons inhibited the PCR product from 10(3) to 10(7) heat-treated cells but could detect 10(1) live L. monocytogenes cells. Our method is useful in clinical as well as food hygiene tests.
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Registered trials
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.