<b>PCR-dipstick DNA chromatography for profiling of a subgroup of cariesassociated bacterial species in plaque from healthy coronal surfaces and periodontal </b><b>pockets </b>

Abstract

The onset of plaque-mediated disease, including dental caries and periodontal diseases, is highly associated with compositional change of the resident microflora from the ecological perspective. As specific bacterial profiles have been linked to different disease stages, microbial compositional measurements might therefore have great value for clinical diagnosis. Previously we have reported a dry-reagent strip biosensor-PCR-dipstick DNA chromatography, which utilized molecular recognition of oligonucleotides and biotin-streptavidin, and the optical property of colored microspheres, for semiquantifying a five-membered subgroup of caries-associated bacterial species in supragingival plaque from healthy coronal surfaces of teeth. The present study aimed to evaluate this technique's ability to differentiate microflora by comparing the subset profiles. Sixteen subgingival plaque specimens were pooled from periodontal pockets and analyzed for the composition of Streptococcus mutans, Streptococcus sobrinus, Scardovia wiggsiae, Actinomyces sp. and Veillonella parvula. Detection frequencies, relative abundance of each bacterial species, and the five-membered bacterial profiles were compared between supra-and subgingival groups. The supragingival plaque harbored significantly more of the tested species and higher amount of Actinomyces sp. and V. parvula. In subgingival plaque, the predominance was obscured, since several highly overlapped profiles were found at comparable frequencies. Thus, PCR-dipstick DNA chromatography using the same plaque sample enabled simultaneous profiling of multiple species at species level and facilitated discrimination between anticipated different microflora, making this technique a promising chair-side microbiota profiling method.The ecological plaque hypothesis (18) has revolutionized our view of the etiology of the most prevalent oral diseases, including caries and periodontal diseases. Contrary to the classical medical model, which ascribes an infection to a single microbial pathogene, the new hypothesis suggests that a dysbiosis of the resident oral microflora, driven by an ecological perturbation, leads to plaque-mediated disease. During the transition from oral health to disease state, a dynamic interaction between the resident microflora and the habitat has been evidenced, with numerous bacterial species being implicated (2,3,22,31). A substantial change of the environmental determinants could disrupt the homeostasis of the resident microflora, thereby inducing the prolifera-

High-resolution microbiome profiling uncoversFusobacterium nucleatum,Lactobacillus gasseri/johnsonii, andLactobacillus vaginalisassociated to oral and oropharyngeal cancer in saliva from HPV positive and HPV negative patients treated with surgery and chemo-radiation

“…Early studies suggest that that the main carbon metabolic pathways, such as the Embden-Meyerhof-Parnas (EMP) pathway, the pentose phosphate (PP) pathway, and the tri-carbonic acid (TCA) cycle, as well as amino acid metabolism-related pathways, are in operation in the oral biofilm in vivo and exhibit similar characteristics to those of Streptococcus and Actinomyces [ 81 , 82 ]. Dysregulation of these main carbon metabolic pathways have been correlated to dental caries, periodontal disease and oral cancer in small studies [ 83 – 85 ]. The following five saliva metabolomic biomarkers can discriminate from early HNSCC patients from controls: propionylcholine, N-Acetyl-L-phenylalanine, sphinganine, phytosphingosine, and S-carboxymethyl-L-cysteine [ 86 ].…”

Development of a Novel Detection Method Targeting an Ultrashort 25 bp Sequence Found in Agrobacterium-Mediated Transformed GM Plants

“…After the PCR or loop-mediated isothermal amplification (LAMP), the C-PAS is dipped into the reaction mixture. The amplification products immobilized with single-tag hybridization are then visually detected with a streptavidin–biotin interaction using a streptavidin-coated blue-colored latex microsphere. − It was revealed that the STH C-PAS system is available for US sequence detection. The LODs determined for each GM event are listed in Table , and the representative results obtained from GM soybeans are shown in Figure .…”

The future of microbiome analysis: Biosensor methods for big data collection and clinical diagnostics

“…Oligonucleotides corresponding to five different bacteria were immobilized at different lines on a paper strip, and due to the attachment of blue latex microspheres to the PCR amplicons, the corresponding test lines became visibly blue after dipping the strip into bacteria‐containing samples (Tian et al., 2014). In 2016, the research group compared the results of their method between the previous healthy individuals and patients with periodontitis, demonstrating reduced microbial diversity in diseased patients as well as differentiation between health‐associated and disease‐associated bacteria (Tian et al., 2016). Methods such as this show the promise for biosensors to be clinically translated for microbiome analysis at the bedside.…”