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- Baumgartner, D, et al.
(author)
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OralDisk: A Chair-Side Compatible Molecular Platform Using Whole Saliva for Monitoring Oral Health at the Dental Practice
- 2021
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In: Biosensors. - : MDPI AG. - 2079-6374. ; 11:11
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Journal article (peer-reviewed)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 ten bacteria: seven periodontitis-associated (Aggregatibacter actinomycetemcomitans, Campylobacter rectus, Fusobacterium nucleatum, Prevotella intermedia, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola) and three caries-associated (oral Lactobacilli, Streptococcus mutans, Streptococcus sobrinus). Each OralDisk test required 400 µL of homogenized whole saliva. The automated workflow included bacterial DNA extraction, purification and hydrolysis probe real-time PCR detection of the target pathogens. All reagents were pre-stored within the disk and sample-to-answer processing took < 3 h using a compact, customized processing device. A technical feasibility study (25 OralDisks) was conducted using samples from healthy, periodontitis and caries patients. The comparison of the OralDisk with a lab-based reference method revealed a ~90% agreement amongst targets detected as positive and negative. This shows the OralDisk’s potential and suitability for inclusion in larger prospective implementation studies in dental care settings.
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- Johannsen, B, et al.
(author)
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Automated Pre-Analytic Processing of Whole Saliva Using Magnet-Beating for Point-of-Care Protein Biomarker Analysis
- 2019
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In: Micromachines. - : MDPI AG. - 2072-666X. ; 10:12
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Journal article (peer-reviewed)abstract
- Saliva offers many advantages for point-of-care (PoC) diagnostic applications due to non-invasive, easy, and cost-effective methods of collection. However, the complex matrix with its non-Newtonian behavior and high viscosity poses handling challenges. Several tedious and long pre-analytic steps, incompatible with PoC use, are required to liquefy and homogenize saliva samples before protein analysis can be performed. We apply magnet-beating to reduce hands-on time and to simplify sample preparation. A magnet in a chamber containing the whole saliva is actuated inside a centrifugal microfluidic cartridge by the interplay of centrifugal and magnetic forces. Rigorous mixing, which homogenizes the saliva sample, is then initiated. Consequently, fewer manual steps are required to introduce the whole saliva into the cartridge. After 4 min of magnet-beating, the processed sample can be used for protein analysis. The viscosity of whole saliva has been reduced from 10.4 to 2.3 mPa s. Immunoassay results after magnet-beating for three salivary periodontal markers (MMP-8, MMP-9, TIMP-1) showed a linear correlation with a slope of 0.99 when compared to results of reference method treated samples. Conclusively, magnet-beating has been shown to be a suitable method for the pre-analytic processing of whole saliva for fully automated PoC protein analysis.
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- Johannsen, B, et al.
(author)
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ImmunoDisk-A Fully Automated Bead-Based Immunoassay Cartridge with All Reagents Pre-Stored
- 2022
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In: Biosensors. - : MDPI AG. - 2079-6374. ; 12:6
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Journal article (peer-reviewed)abstract
- 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.
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- Milone, A. P., et al.
(author)
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The ACS survey of Galactic globular clusters XII. Photometric binaries along the main sequence
- 2012
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In: Astronomy & Astrophysics. - : EDP Sciences. - 0004-6361 .- 1432-0746. ; 540
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Journal article (peer-reviewed)abstract
- Context. The fraction of binary stars is an important ingredient to interpret globular cluster dynamical evolution and their stellar population. Aims. We investigate the properties of main-sequence binaries measured in a uniform photometric sample of 59 Galactic globular clusters that were observed by HST WFC/ACS as a part of the Globular Cluster Treasury project. Methods. We measured the fraction of binaries and the distribution of mass-ratio as a function of radial location within the cluster, from the central core to beyond the half-mass radius. We studied the radial distribution of binary stars, and the distribution of stellar mass ratios. We investigated monovariate relations between the fraction of binaries and the main parameters of their host clusters. Results. We found that in nearly all the clusters, the total fraction of binaries is significantly smaller than the fraction of binaries in the field, with a few exceptions only. Binary stars are significantly more centrally concentrated than single MS stars in most of the clusters studied in this paper. The distribution of the mass ratio is generally flat (for mass-ratio parameter q > 0.5). We found a significant anti-correlation between the binary fraction in a cluster and its absolute luminosity (mass). Some, less significant correlation with the collisional parameter, the central stellar density, and the central velocity dispersion are present. There is no statistically significant relation between the binary fraction and other cluster parameters. We confirm the correlation between the binary fraction and the fraction of blue stragglers in the cluster.
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