Abstract:Periodontitis and dental caries are two major bacterially induced, non-communicable diseases that cause the deterioration of oral health, with implications in patients’ general health. Early, precise diagnosis and personalized monitoring are essential for the efficient prevention and management of these diseases. Here, we present a disk-shaped microfluidic platform (OralDisk) compatible with chair-side use that enables analysis of non-invasively collected whole saliva samples and molecular-based detection of t… Show more
“…Lyophilization or freeze-drying is often used to store proteins (alone, i.e., without beads) due to the good stabilization achieved with this method, even though it is costly and technologically complex [ 61 ]. Concerning beads alone, we have demonstrated protocols for pre-storage in the case of magnetic beads for nucleic acid extraction and purification [ 62 , 63 , 64 ]. However, the BFPD-IA that is integrated within the ImmunoDisk involves protein-coupled magnetic and fluorescent beads.…”
Section: Resultsmentioning
confidence: 99%
“…These conditions simplify the fluidic integration and reduce the footprint on disk. Importantly, the ImmunoDisk was processed on a device, the LabDisk Player 1 functional model, which was developed for the automation of nucleic acid amplification technologies (NAATs, e.g., PCR and isothermal methods) and for which different applications have already been shown (e.g., respiratory tract infections [ 64 ], tropical infections [ 63 ], oral diseases [ 62 ] and vector analysis in mosquitos [ 76 ]), without any changes to the device hardware. The compatibility of the ImmunoDisk with this NAAT device is primarily due to the protocol and the detection principle of the BFPD-IA, as well as the adaptation of the microfluidic structures.…”
In this paper, we present the ImmunoDisk, a fully automated sample-to-answer centrifugal microfluidic cartridge, integrating a heterogeneous, wash-free, magnetic- and fluorescent bead-based immunoassay (bound-free phase detection immunoassay/BFPD-IA). The BFPD-IA allows the implementation of a simple fluidic structure, where the assay incubation, bead separation and detection are performed in the same chamber. The system was characterized using a C-reactive protein (CRP) competitive immunoassay. A parametric investigation on air drying of protein-coupled beads for pre-storage at room temperature is presented. The key parameters were buffer composition, drying temperature and duration. A protocol for drying two different types of protein-coupled beads with the same temperature and duration using different drying buffers is presented. The sample-to-answer workflow was demonstrated measuring CRP in 5 µL of human serum, without prior dilution, utilizing only one incubation step, in 20 min turnaround time, in the clinically relevant concentration range of 15–115 mg/L. A reproducibility assessment over three disk batches revealed an average signal coefficient of variation (CV) of 5.8 ± 1.3%. A CRP certified reference material was used for method verification with a concentration CV of 8.6%. Our results encourage future testing of the CRP-ImmunoDisk in clinical studies and its point-of-care implementation in many diagnostic applications.
“…Lyophilization or freeze-drying is often used to store proteins (alone, i.e., without beads) due to the good stabilization achieved with this method, even though it is costly and technologically complex [ 61 ]. Concerning beads alone, we have demonstrated protocols for pre-storage in the case of magnetic beads for nucleic acid extraction and purification [ 62 , 63 , 64 ]. However, the BFPD-IA that is integrated within the ImmunoDisk involves protein-coupled magnetic and fluorescent beads.…”
Section: Resultsmentioning
confidence: 99%
“…These conditions simplify the fluidic integration and reduce the footprint on disk. Importantly, the ImmunoDisk was processed on a device, the LabDisk Player 1 functional model, which was developed for the automation of nucleic acid amplification technologies (NAATs, e.g., PCR and isothermal methods) and for which different applications have already been shown (e.g., respiratory tract infections [ 64 ], tropical infections [ 63 ], oral diseases [ 62 ] and vector analysis in mosquitos [ 76 ]), without any changes to the device hardware. The compatibility of the ImmunoDisk with this NAAT device is primarily due to the protocol and the detection principle of the BFPD-IA, as well as the adaptation of the microfluidic structures.…”
In this paper, we present the ImmunoDisk, a fully automated sample-to-answer centrifugal microfluidic cartridge, integrating a heterogeneous, wash-free, magnetic- and fluorescent bead-based immunoassay (bound-free phase detection immunoassay/BFPD-IA). The BFPD-IA allows the implementation of a simple fluidic structure, where the assay incubation, bead separation and detection are performed in the same chamber. The system was characterized using a C-reactive protein (CRP) competitive immunoassay. A parametric investigation on air drying of protein-coupled beads for pre-storage at room temperature is presented. The key parameters were buffer composition, drying temperature and duration. A protocol for drying two different types of protein-coupled beads with the same temperature and duration using different drying buffers is presented. The sample-to-answer workflow was demonstrated measuring CRP in 5 µL of human serum, without prior dilution, utilizing only one incubation step, in 20 min turnaround time, in the clinically relevant concentration range of 15–115 mg/L. A reproducibility assessment over three disk batches revealed an average signal coefficient of variation (CV) of 5.8 ± 1.3%. A CRP certified reference material was used for method verification with a concentration CV of 8.6%. Our results encourage future testing of the CRP-ImmunoDisk in clinical studies and its point-of-care implementation in many diagnostic applications.
“…These molecular platforms enable a very rapid sampling-to-answer pipeline (i.e., within a patient session). Chair-side assays under development include molecular quantification of periodontal or cariogenic species [ 40 – 42 ] or the detection of antibiotic resistance genes [ 43 ] within oral samples.…”
Diagnosis and treatment in dentistry are based on clinical examination of the patients. Given that the major oral diseases are of microbial biofilm etiology, it can be expected that performing microbiological analysis on samples collected from the patient could deliver supportive evidence to facilitate the decision-making process by the clinician. Applicable microbiological methods range from microscopy, to culture, to molecular techniques, which can be performed easily within dedicated laboratories proximal to the clinics, such as ones in academic dental institutions. Periodontal and endodontic infections, along with odontogenic abscesses, have been identified as conditions in which applied clinical microbiology may be beneficial for the patient. Administration of antimicrobial agents, backed by microbiological analysis, can yield more predictable treatment outcomes in refractory or early-occurring forms of periodontitis. Confirming a sterile root canal using a culture-negative sample during endodontic treatment may ensure the longevity of its outcome and prevent secondary infections. Susceptibility testing of samples obtained from odontogenic abscesses may facilitate the selection of the appropriate antimicrobial treatment to prevent further spread of the infection.
“…Aside, it seems crucial to control and detect individuals' deviations of the oral health status at an early stage to hinder disease outbreak. While gold standard clinical examinations and radiographs at the dental office may miss developing disorders on a cellular base, other diagnostic tools and modern technical possibilities gain more importance ( 4 , 5 ). Different biofluids, such as periphery blood, gingival crevicular fluid (GCF), and saliva were evaluated for their diagnostic or prognostic qualification for disease detection ( 6 ).…”
Section: Introductionmentioning
confidence: 99%
“…It was shown, that the oral microbiome can be quantified to a certain extent in saliva ( 9 , 10 ), and even more interesting, host immune response to pathogens can be analyzed nearly in real-time ( 11 ). Furthermore, modern technologies allow the detection and quantification of specific substances in low concentrations ( 4 , 12 ). Today, improvements and further developments in point-of-care devices evolve from technical progress in the era of Covid 19 ( 13 , 14 ).…”
Personalized Oral Healthcare has recently become the new trend word in medicine and dentistry. In this context, saliva diagnostics using various biomarkers seem to be the gateway to personalized dental diagnostics and therapy. But the terminology is not (yet) uniformly defined, furthermore it is unclear to what extent which salivary markers play a relevant role in the therapeutic decision making. In this Scoping Review, an electronic search was conducted in PubMed and Web of Science databases using medical subject headings (MESH terms) “saliva”, “biomarker”, “personality/persons”, and “dentistry”. Only human studies were included, in which repeated salivary measurements were performed to analyze monitoring effects with at least ten patients per group. PRISMA-ScR and Tricco guidelines were followed: (i) to examine what salivary biomarkers have been explored in terms of personalized oral healthcare and precision dentistry, (ii) to investigate the clinical relevance for oral health and its correlation to systemic health, and (iii) to summarize an outlook for future developments based on these results. Out of 899 studies, a total of 57 were included for data extraction in this Scoping Review, mainly focusing on periodontal therapy and patient monitoring. Salivary biomarkers have shown the potential to change the field of dentistry in all dental disciplines as a key for personalized workflows. The increasing interest in dental research is obvious, demonstrated by the growing number of publications in recent years. At this time, however, the predominant discipline is periodontology, which allows biomarker-based monitoring of the disease prevention and progression. The studies included showed heterogeneous methods using manifolds biomarkers. Therefore, no uniformly accepted concept can be presented today. Further clinical research with well-defined outcomes including standardized procedures is necessary.
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