ArticleMicrobiology spectrum2022
Absolute Quantification of Viable but Nonculturable Vibrio cholerae Using Droplet Digital PCR with Oil-Enveloped Bacterial Cells.
Article in Microbiology spectrum, 2022. 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, 19 citations in OpenAlex.
- Revisiting Strategies for Bacterial Enumeration: The Case for Viable but Nonculturable (VBNC) Cells in Supplement Products.Probiotics and antimicrobial proteins · 2026Review
- Absolute quantification of VBNC state formation and resuscitation in alcohol-producingFrontiers in cellular and infection microbiology · 2025Article
- Development of Recombinant Antibodies and Its Application in Immunomagnetic Separation-Based Rapid Detection ofJournal of microbiology and biotechnology · 2024Article
- Persistence and Culturability ofAntibiotics (Basel, Switzerland) · 2024Article
- Multiple factors trigger the formation and resuscitation of the VBNC state in alcohol-producingApplied and environmental microbiology · 2024Article
- Public health aspects ofEFSA journal. European Food Safety Authority · 2024Article
- Development and evaluation of a multi-target droplet digital PCR assay for highly sensitive and specific detection of Yersinia pestis.PLoS neglected tropical diseases · 2024Article
- Pathways of exposure to Vibrio Cholerae in an urban informal settlement in Nairobi, Kenya.PLOS global public health · 2024Article
- Cross-Platform Transcriptomic Data Integration, Profiling, and Mining inMicrobiology spectrum · 2023Article
- Probiotic and postbiotic analytical methods: a perspective of available enumeration techniques.Frontiers in microbiology · 2023Review
- Viability droplet digital polymerase chain reaction accurately enumerates probiotics and provides insight into damage experienced during storage.Frontiers in microbiology · 2022Article
- A triton X-100 assisted PMAxx-qPCR assay for rapid assessment of infectious African swine fever virus.Frontiers in microbiology · 2022Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
When exposed to adverse conditions, many bacterial pathogens, including Vibrio cholerae, can adapt to the environment by entering the viable but nonculturable (VBNC) state. Cells in this state cannot grow on conventional media but still survive. The VBNC state is a significant threat to aquatic safety and public health due to the enhanced ability of the bacteria to remain in the environment and escape from monitoring. Detecting and quantifying VBNC cells and distinguishing them from nonviable cells is necessary for microbiological studies and pathogen monitoring. Cell staining and microscopy are used for the observation of VBNC cells, but it is difficult to quantify VBNC cells accurately. In this study, we developed droplet digital PCR (ddPCR) with a chromosomal single-copy gene as an internal reference combined with Propidium monoazide (PMA) treatment to enumerate VBNC cells of V. cholerae. In this method, bacterial cells, but not extracted chromosomal DNA, were used directly to form oil-enveloped droplets in the ddPCR procedure. One bacterial cell possesses one copy of the chromosome. Thus, enumeration of a single-copy gene on the chromosome can be used to count VBNC cells. ddPCR showed greater accuracy and sensitivity than qPCR. This study showed that the oil-enveloped bacterial method can reduce the number of steps needed and improve the accuracy of VBNC cells quantification and has the potential to be extended to quantify bacterial VBNC cells and assess pathogen survival in the environment.
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Registered trials
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