scholarly journals Rapid and Sensitive PCR-Dipstick DNA Chromatography for Multiplex Analysis of the Oral Microbiota

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Lingyang Tian ◽  
Takuichi Sato ◽  
Kousuke Niwa ◽  
Mitsuo Kawase ◽  
Anne C. R. Tanner ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Dental Plaque ◽  
Bacterial Species ◽  
Oral Microbiota ◽  
Streptococcus Sobrinus ◽  
Microbial Composition ◽  
Test Species ◽  
Multiplex Analysis ◽  
Latex Beads ◽  
Target Dna

A complex of species has been associated with dental caries under the ecological hypothesis. This study aimed to develop a rapid, sensitive PCR-dipstick DNA chromatography assay that could be read by eye for multiplex and semiquantitative analysis of plaque bacteria. Parallel oligonucleotides were immobilized on a dipstick strip for multiplex analysis of target DNA sequences of the caries-associated bacteria,Streptococcus mutans,Streptococcus sobrinus,Scardovia wiggsiae,Actinomycesspecies, andVeillonella parvula. Streptavidin-coated blue-colored latex microspheres were to generate signal. Target DNA amplicons with an oligonucleotide-tagged terminus and a biotinylated terminus were coupled with latex beads through a streptavidin-biotin interaction and then hybridized with complementary oligonucleotides on the strip. The accumulation of captured latex beads on the test and control lines produced blue bands, enabling visual detection with the naked eye. The PCR-dipstick DNA chromatography detected quantities as low as 100 pg of DNA amplicons and demonstrated 10- to 1000-fold higher sensitivity than PCR-agarose gel electrophoresis, depending on the target bacterial species. Semiquantification of bacteria was performed by obtaining a series of chromatograms using serial 10-fold dilution of PCR-amplified DNA extracted from dental plaque samples. The assay time was less than 3 h. The semiquantification procedure revealed the relative amounts of each test species in dental plaque samples, indicating that this disposable device has great potential in analysis of microbial composition in the oral cavity and intestinal tract, as well as in point-of-care diagnosis of microbiota-associated diseases.

scholarly journals Detection of streptococcus mutans and streptococcus sobrinus in dental plaque samples from Brazilian preschool children by polymerase chain reaction

2007 ◽  
Vol 18 (4) ◽  
pp. 329-333 ◽  
Author(s):  
Teresa Cristina Costa Franco e Franco ◽  
Patrícia Amoroso ◽  
José Moacir Marin ◽  
Fernando Antonio de Ávila
Keyword(s):  
Polymerase Chain Reaction ◽  
Preschool Children ◽  
Dental Plaque ◽  
Bacterial Species ◽  
Mutans Streptococci ◽  
World Health ◽  
Streptococcus Sobrinus ◽  
Chain Reaction ◽  
Discriminative Ability ◽  
Polymerase Chain

The purposes of this study were to detect S. mutans and S. sobrinus by polymerase chain reaction (PCR) amplification, and to relate their presence to the incidence of dental caries in 42 Brazilian preschool children. Dental plaque samples were collected from the cervical margin of all erupted teeth of 5-6 years old children with primary dentition, using a sterile explorer. Examination of the dmft (decayed, missing, filled teeth) index, performed following the World Health Organization (WHO) caries diagnostic criteria, showed a 2.71 score. Prevalence of S. mutans and S. sobrinus was respectively, of 85.7% and 14.3%; no dental plaque sample was either positive or negative for both bacterial species. Children harboring either S. mutans or S. sobrinus presented the same caries prevalence. PCR showed good discriminative ability for differentiation between these species, and suggested that it is a technique suitable for epidemiological studies on mutans streptococci.

scholarly journals Oral Microbiota Composition and Function Changes During Chronic Erythematous Candidiasis

2021 ◽  
Vol 11 ◽  
Author(s):  
Xin Lyu ◽  
Hui Zheng ◽  
Xu Wang ◽  
Heyu Zhang ◽  
Lu Gao ◽  
...  
Keyword(s):  
Healthy Subjects ◽  
Bacterial Species ◽  
Oral Bacteria ◽  
Oral Microbiome ◽  
Marker Genes ◽  
Oral Microbiota ◽  
Metagenomic Sequencing ◽  
Cytochrome Bc1 Complex ◽  
Microbial Composition ◽  
Treatment And Prevention

Oral microbiota is constantly changing with the host state, whereas the oral microbiome of chronic erythematous candidiasis remains poorly understood. The aim of this study was to compare oral microbial signatures and functional profiling between chronic erythematous candidiasis and healthy subjects. Using shotgun metagenomic sequencing, we analyzed the microbiome in 12 chronic erythematous candidiasis, 12 healthy subjects, and 2 chronic erythematous candidiasis cured by antifungal therapy. We found that the salivary microbiota of chronic erythematous candidiasis was significantly different from that of healthy subjects. Among them, Rothia mucilaginosa and Streptococcus mitis were the most abundant disease-enriched species (Mann-Whitney U-test, P < 0.05). In addition, co-occurrence network analysis showed that C. albicans formed densely connected modules with oral bacterial species and was mainly positive connected to Streptococcus species. Furthermore, we investigated the functional potentials of the microbiome and identified a set of microbial marker genes associated with chronic erythematous candidiasis. Some of these genes enriching in chronic erythematous candidiasis are involved in eukaryotic ribosome, putative glutamine transport system, and cytochrome bc1 complex respiratory unit. Altogether, this study revealed the changes of oral microbial composition, the co-occurrence between C. albicans and oral bacteria, as well as the changes of microbial marker genes during chronic erythematous candidiasis, which provides evidence of oral microbiome as a target for the treatment and prevention of chronic erythematous candidiasis.

scholarly journals Maturation of the Oral Microbiome in Caries-Free Toddlers: A Longitudinal Study

2019 ◽  
Vol 99 (2) ◽  
pp. 159-167 ◽  
Author(s):  
D. Kahharova ◽  
B.W. Brandt ◽  
M.J. Buijs ◽  
M. Peters ◽  
R. Jackson ◽  
...  
Keyword(s):  
Dental Plaque ◽  
Illumina Miseq ◽  
Assessment System ◽  
Oral Microbiome ◽  
Caries Detection ◽  
Oral Microbiota ◽  
Healthy Children ◽  
Microbial Composition ◽  
Time Points ◽  
Over Time

Understanding the development of the oral microbiota in healthy children is of great importance to oral and general health. However, limited data exist on a healthy maturation of the oral microbial ecosystem in children. Moreover, the data are biased by mislabeling “caries-free” populations. Therefore, we aimed to characterize the healthy salivary and dental plaque microbiome in young children. Caries-free (ICDAS [International Caries Detection and Assessment System] score 0) children ( n = 119) and their primary caregivers were followed from 1 until 4 y of child age. Salivary and dental plaque samples were collected from the children at 3 time points (T1, ~1 y old; T2, ~2.5 y old; and T3, ~4 y old). Only saliva samples were collected from the caregivers. Bacterial V4 16S ribosomal DNA amplicons were sequenced using Illumina MiSeq. The reads were denoised and mapped to the zero-radius operational taxonomic units (zOTUs). Taxonomy was assigned using HOMD. The microbial profiles of children showed significant differences ( P = 0.0001) over time. Various taxa increased, including Fusobacterium, Actinomyces, and Corynebacterium, while others showed significant decreases (e.g., Alloprevotella and Capnocytophaga) in their relative abundances over time. Microbial diversity and child-caregiver similarity increased most between 1 and 2.5 y of age while still not reaching the complexity of the caregivers at 4 y of age. The microbiome at 1 y of age differed the most from those at later time points. A single zOTU ( Streptococcus) was present in all samples ( n = 925) of the study. A large variation in the proportion of shared zOTUs was observed within an individual child over time (2% to 42% of zOTUs in saliva; 2.5% to 38% in dental plaque). These findings indicate that the oral ecosystem of caries-free toddlers is highly heterogeneous and dynamic with substantial changes in microbial composition over time and only few taxa persisting across the 3 y of the study. The salivary microbiome of 4-y-old children is still distinct from that of their caregivers.

scholarly journals Association of Bitter Taste Receptor T2R38 Polymorphisms, Oral Microbiota, and Rheumatoid Arthritis

2021 ◽  
Vol 43 (3) ◽  
pp. 1460-1472
Author(s):  
Vivianne Cruz de Jesus ◽  
Manu Singh ◽  
Robert J. Schroth ◽  
Prashen Chelikani ◽  
Carol A. Hitchon
Keyword(s):  
Rheumatoid Arthritis ◽  
Relative Abundance ◽  
Bitter Taste ◽  
Bacterial Species ◽  
Taste Receptor ◽  
Oral Microbiome ◽  
Rrna Gene ◽  
Oral Microbiota ◽  
Microbial Composition ◽  
The Impact

The association of taste genetics and the oral microbiome in autoimmune diseases such as rheumatoid arthritis (RA) has not been reported. We explored a novel oral mucosal innate immune pathway involving the bitter taste G protein-coupled receptor T2R38. This case–control study aimed to evaluate whether T2R38 polymorphisms associate with the buccal microbial composition in RA. Genomic DNA was obtained from buccal swabs of 35 RA patients and 64 non-RA controls. TAS2R38 genotypes were determined by Sanger sequencing. The buccal microbiome was assessed by Illumina MiSeq sequencing of the V4-16S rRNA gene. Bacterial community differences were analyzed with alpha and beta diversity measures. Linear discriminant analysis effect size identified taxa discriminating between RA versus non-RA and across TAS2R38 genotypes. TAS2R38 genotype frequency was similar between RA and non-RA controls (PAV/PAV; PAV/AVI; AVI/AVI: RA 42.9%; 45.7%; 11.4% versus controls 32.8%; 48.4%; 18.8%, chi-square (2, N = 99) = 2.1, p = 0.35). The relative abundance of Porphyromonas, among others, differed between RA and non-RA controls. The relative abundance of several bacterial species also differed across TAS2R38 genotypes. These findings suggest an association between T2R38 polymorphisms and RA buccal microbial composition. However, further research is needed to understand the impact of T2R38 in oral health and RA development.

scholarly journals Evaluation of Changes in Cariogenic Bacteria in a Young Moroccan Population with Fixed Orthodontic Appliances

2018 ◽  
Vol 2018 ◽  
pp. 1-4
Author(s):  
A. Marda ◽  
S. Elhamzaoui ◽  
A. El Mansari ◽  
K. Souly ◽  
F. Farissi ◽  
...  
Keyword(s):  
Dental Plaque ◽  
Orthodontic Treatment ◽  
Bacterial Species ◽  
Qualitative Change ◽  
Orthodontic Appliances ◽  
Streptococcus Sobrinus ◽  
Vitamin K3 ◽  
Moroccan Population ◽  
Fixed Orthodontic Appliances ◽  
Significant Difference

Fixed orthodontic appliances hinder the maintenance of proper oral hygiene and result in dental plaque accumulation. Many studies report that qualitative changes in the dental flora occur after initiating the orthodontic treatment, but there is a paucity of literature on the same topic among Moroccan orthodontic patients. The aim of this study was to evaluate the changes of the oral microbial flora during the orthodontic treatment period of a young Moroccan population. Materials and Methods. Dental plaque samples of 18 patients, who were randomly selected before the placement of orthodontic appliances, were collected to isolate and identify the bacterial species involved using classical bacteriological methods for species’ culture and identification. The reading was recorded at T0 before placement of the device. New samples were taken again one month later and then three months afterwards, where the readings were recorded as T1 and T2, respectively. The culture was made via Columbia Agar with 5% sheep blood, Todd Hewitt Broth, and Schaedler medium containing vitamin K3. Bacterial species were identified using API-20 Strep for Streptococci and API-20 A for anaerobic bacteria. The phoenix system was used for identification. Results. After three months of orthodontic treatment, the increase in the frequency of Streptococcus sobrinus and Streptococcus mitis were significant (0.01 and 0.02, respectively) as well as for Lactobacillus (0.03). No significant difference was recorded for other bacterial species. Conclusion. There is a significant qualitative change in oral microorganisms after three months of orthodontic treatment, especially for bacteria that are incriminated in caries formation.

scholarly journals Evaluation of acidogenesis products’ effect on biogas production performed with metagenomics and isotopic approaches

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Anna Detman ◽  
Michał Bucha ◽  
Laura Treu ◽  
Aleksandra Chojnacka ◽  
Łukasz Pleśniak ◽  
...  
Keyword(s):  
Microbial Communities ◽  
Methane Production ◽  
Bacterial Species ◽  
Stable Carbon Isotope ◽  
Anaerobic Sludge ◽  
Methane Formation ◽  
Microbial Composition ◽  
Fermentation Products ◽  
Core Microbiome ◽  
Specific Species

Abstract Background During the acetogenic step of anaerobic digestion, the products of acidogenesis are oxidized to substrates for methanogenesis: hydrogen, carbon dioxide and acetate. Acetogenesis and methanogenesis are highly interconnected processes due to the syntrophic associations between acetogenic bacteria and hydrogenotrophic methanogens, allowing the whole process to become thermodynamically favorable. The aim of this study is to determine the influence of the dominant acidic products on the metabolic pathways of methane formation and to find a core microbiome and substrate-specific species in a mixed biogas-producing system. Results Four methane-producing microbial communities were fed with artificial media having one dominant component, respectively, lactate, butyrate, propionate and acetate, for 896 days in 3.5-L Up-flow Anaerobic Sludge Blanket (UASB) bioreactors. All the microbial communities showed moderately different methane production and utilization of the substrates. Analyses of stable carbon isotope composition of the fermentation gas and the substrates showed differences in average values of δ13C(CH4) and δ13C(CO2) revealing that acetate and lactate strongly favored the acetotrophic pathway, while butyrate and propionate favored the hydrogenotrophic pathway of methane formation. Genome-centric metagenomic analysis recovered 234 Metagenome Assembled Genomes (MAGs), including 31 archaeal and 203 bacterial species, mostly unknown and uncultivable. MAGs accounted for 54%–67% of the entire microbial community (depending on the bioreactor) and evidenced that the microbiome is extremely complex in terms of the number of species. The core microbiome was composed of Methanothrix soehngenii (the most abundant), Methanoculleus sp., unknown Bacteroidales and Spirochaetaceae. Relative abundance analysis of all the samples revealed microbes having substrate preferences. Substrate-specific species were mostly unknown and not predominant in the microbial communities. Conclusions In this experimental system, the dominant fermentation products subjected to methanogenesis moderately modified the final effect of bioreactor performance. At the molecular level, a different contribution of acetotrophic and hydrogenotrophic pathways for methane production, a very high level of new species recovered, and a moderate variability in microbial composition depending on substrate availability were evidenced. Propionate was not a factor ceasing methane production. All these findings are relevant because lactate, acetate, propionate and butyrate are the universal products of acidogenesis, regardless of feedstock.

scholarly journals Pre and Probiotics Involved in the Modulation of Oral Bacterial Species: New Therapeutic Leads in Mental Disorders?

2021 ◽  
Vol 9 (7) ◽  
pp. 1450
Author(s):  
Yoann Maitre ◽  
Rachid Mahalli ◽  
Pierre Micheneau ◽  
Alexis Delpierre ◽  
Marie Guerin ◽  
...  
Keyword(s):  
Systematic Review ◽  
Central Nervous System ◽  
Nervous System ◽  
Oral Health ◽  
Mental Disorders ◽  
Meta Analysis ◽  
Bacterial Species ◽  
Oral Microbiome ◽  
Oral Microbiota ◽  
Therapeutic Leads

This systematic review aims to identify probiotics and prebiotics for modulating oral bacterial species associated with mental disorders. Using the Preferred Reporting Items for Systematic Reviews and Meta-Analysis guideline, we search the electronic MEDLINE database published till January 2021 to identify the studies on probiotics and/or prebiotics for preventing and treating major oral dysbiosis that provokes mental disorders. The outcome of the search produces 374 records. After excluding non-relevant studies, 38 papers were included in the present review. While many studies suggest the potential effects of the oral microbiota on the biochemical signalling events between the oral microbiota and central nervous system, our review highlights the limited development concerning the use of prebiotics and/or probiotics in modulating oral dysbiosis potentially involved in the development of mental disorders. However, the collected studies confirm prebiotics and/or probiotics interest for a global or targeted modulation of the oral microbiome in preventing or treating mental disorders. These outcomes also offer exciting prospects for improving the oral health of people with mental disorders in the future.

scholarly journals Epithelial cell pro-inflammatory cytokine response differs across dental plaque bacterial species

2010 ◽  
Vol 37 (1) ◽  
pp. 24-29 ◽  
Author(s):  
Panagiota G. Stathopoulou ◽  
Manjunatha R. Benakanakere ◽  
Johnah C. Galicia ◽  
Denis F. Kinane
Keyword(s):  
Epithelial Cell ◽  
Dental Plaque ◽  
Inflammatory Cytokine ◽  
Bacterial Species ◽  
Cytokine Response ◽  
Inflammatory Cytokine Response

A high sensitivity wireless mass-loading surface acoustic wave DNA biosensor

2014 ◽  
Vol 28 (07) ◽  
pp. 1450056 ◽  
Author(s):  
Hua-Lin Cai ◽  
Yi Yang ◽  
Yi-Han Zhang ◽  
Chang-Jian Zhou ◽  
Cang-Ran Guo ◽  
...  
Keyword(s):  
Acoustic Wave ◽  
Surface Acoustic Wave ◽  
Dna Sequences ◽  
Biological Treatment ◽  
High Performance ◽  
Processing System ◽  
High Sensitivity ◽  
Treatment Method ◽  
Saw Sensor ◽  
Target Dna

In this paper, a surface acoustic wave (SAW) biosensor with gold delay area on LiNbO 3 substrate detecting DNA sequences is proposed. By well-designed device parameters of the SAW sensor, it achieves a high performance for highly sensitive detection of target DNA. In addition, an effective biological treatment method for DNA immobilization and abundant experimental verification of the sensing effect have made it a reliable device in DNA detection. The loading mass of the probe and target DNA sequences is obtained from the frequency shifts, which are big enough in this work due to an effective biological treatment. The experimental results show that the biosensor has a high sensitivity of 1.2 pg/ml/Hz and high selectivity characteristic is also verified by the few responses of other substances. In combination with wireless transceiver, we develop a wireless receiving and processing system that can directly display the detection results.

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