| Title |
Method to quantify live and dead cells in multi-species oral biofilm by real-time PCR with propidium monoazide
|
|---|---|
| Published in |
AMB Express, January 2013
|
| DOI | 10.1186/2191-0855-3-1 |
| Pubmed ID | |
| Authors |
Gerard Àlvarez, Marta González, Sergio Isabal, Vanessa Blanc, Rubén León |
| Abstract |
Real-time PCR (qPCR) is a widely used technique in analysing environmental and clinical microbiological samples. However, its main limitation was its inability to discriminate between live and dead cells.Recently, propidium monoazide (PMA) together with qPCR has been used to overcome this problem, with good results for different bacterial species in different types of samples.Our objective was to implement this technique for analysing mortality in multi-species oral biofilms formed in vitro with five oral bacteria: Streptococcus oralis, Streptococcus gordonii, Veillonella parvula, Fusobacterium nucleatum and Prevotella intermedia. We also tested its effectiveness on biofilms treated with an antiseptic solution containing 0.07% w/w cetylpyridinium chloride (CPC).Standardisation of the qPCR-PMA method was performed on pure, heat-killed planktonic cultures of each species, detecting mortality higher than 4 log in S. oralis, S. gordonii and F. nucleatum and higher than 2 for V. parvula and P. intermedia. We obtained similar results for all species when using CPC.When we analysed biofilms with qPCR-PMA, we found that the mortality in the non-CPC treated multi-species biofilms was lower than 1 log for all species. After treatment with CPC, the viability reduction was higher than 4 log in S. oralis and S. gordonii, higher than 3 log in F. nucleatum and P. intermedia and approximately 2 in V. parvula.In short, we standardised the conditions for using qPCR-PMA in 5 oral bacterial species and proved its usefulness for quantification of live and dead cells in multi-species oral biofilms formed in vitro, after use of an antiseptic. |
Mendeley readers
The data shown below were compiled from readership statistics for 147 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 147 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Master | 6 | 4% |
| Student > Ph. D. Student | 5 | 3% |
| Researcher | 4 | 3% |
| Professor | 4 | 3% |
| Student > Bachelor | 4 | 3% |
| Other | 12 | 8% |
| Unknown | 112 | 76% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 9 | 6% |
| Agricultural and Biological Sciences | 6 | 4% |
| Pharmacology, Toxicology and Pharmaceutical Science | 5 | 3% |
| Immunology and Microbiology | 5 | 3% |
| Chemical Engineering | 3 | 2% |
| Other | 7 | 5% |
| Unknown | 112 | 76% |
Attention Score in Context
This research output has an Altmetric Attention Score of 4. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 12 December 2023.
All research outputs
#7,178,663
of 22,691,736 outputs
Outputs from AMB Express
#156
of 1,228 outputs
Outputs of similar age
#80,207
of 280,650 outputs
Outputs of similar age from AMB Express
#5
of 20 outputs
Altmetric has tracked 22,691,736 research outputs across all sources so far. This one has received more attention than most of these and is in the 67th percentile.
So far Altmetric has tracked 1,228 research outputs from this source. They receive a mean Attention Score of 2.8. This one has done well, scoring higher than 85% of its peers.
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We're also able to compare this research output to 20 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 70% of its contemporaries.