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Simple, Label-Free and Non-Instrumented Analyte Quantitation by Flow Distance Measurement in Microfluidic DevicesChatterjee, Debolina 18 August 2014 (has links) (PDF)
Rapid determination of the concentrations of molecules related to diseases can provide timely information for treatment options. However, most biomarker quantitation methods require costly and complex equipment. On the other hand, point-of-care systems have less complex instrumentation needs than laboratory-based equipment, but often provide less information; for example, biomarker presence or absence instead of concentration. A complete analysis setup addressing key limitations of both laboratory-based and portable systems is highly desirable. I developed microfluidic devices with visual inspection readout of a target’s concentration from microliter volumes of solution flowed into a microchannel. Microchannels are formed within polydimethylsiloxane (PDMS), and the surfaces are coated with receptors. Capillary flow of target solution in the channel crosslinks the top and bottom surfaces, which constricts the channel and stops flow. The flow distance of the target solution in the channel before flow stops indicates the target’s concentration, enabling simple visual inspection readout without complex detection instrumentation. Because of its easy readout and portability, my system has great potential for use in point-of-care diagnostics. I initially demonstrated a proof-of-concept assay using biotin-streptavidin. Solution capillary flow distances scaled linearly with the negative logarithm of streptavidin concentration over a 100,000-fold range. I measured streptavidin concentrations as low as 1 ng/mL using these microsystems, demonstrating low detection limits. I also characterized the mechanism wherein time-dependent channel constriction in the first few millimeters leads to concentration-dependent flow distances. I demonstrated the visual detection and quantification capability of my system to determine an antigen target, thymidine kinase 1 (TK1). I developed surface modification methods for carrying out flow assays and verified receptor attachment on channel surfaces using fluorescence imaging. I obtained a 1 ng/mL TK1 detection limit in flow assays. I also demonstrated nucleic acid quantitation in my flow devices. I detected specific DNA targets in buffer and synthetic urine at 10 pg/mL levels. A dynamic range of 106 was obtained with single-base mismatch specificity. DNA analogues of two miRNA biomarkers were measured near clinically significant levels, showing great promise for future medical application. The promising results demonstrate that this diagnostic tool offers a simple route to analyte quantitation in microliter volumes, with excellent potential for point-of-care application.
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Thesis_SP_12062022.pdfSangeeta Pandey (14226758) 08 December 2022 (has links)
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<p>Miniature mass spectrometers are in a phase of rapid development due to their potential in offering simple yet powerful solutions for a wide variety of unmet biomedical needs. In particular, the combination of ambient ionization methods with miniature mass spectrometers offers an attractive solution for improving patient outcomes and reducing the healthcare burden on patients as well as clinicians opposed to current methods for disease prognosis and diagnosis. </p>
<p>There has been a rapid expansion in the commercial offerings of miniature mass spectrometers from commercial vendors, both large and small, including Purspec, Bayspec, MassTech, Waters, and Advion. Despite the large number of instruments that have been made available and the success of many of these systems with analysis of a broad range of biological matrices, much work remains to perform bioanalysis of complex molecules with concentrations that often lie in the ng/mL-µg/mL range. Miniaturization of mass spectrometers is accompanied by design simplifications in comparison to benchtop instruments, so that sacrifices are often made in terms of performance. The figure of merit that is compromised, of course, depends on the design of the instrument itself. Thus, in addition to a good understanding of the operation of the mass spectrometer, it is crucial that the ionization method for the analyte be chosen judiciously, and that the method is suitably optimized to be able to perform the measurements to obtain high quality data for trace analysis. </p>
<p>The custom built Mini-12 miniature MS systems at Purdue University is one such miniature mass spectrometer that can be used for making on-site measurements. In this work, I have described my efforts to perform trace analysis of a range of molecules (tenofovir diphosphate, cabotegravir, rilpivirine, and phosphatidylethanol) relevant to HIV treatment and prevention with the Mini-12 system. Based on the most favorable set of conditions for developing a particular assay, method performance parameters are listed for each of the applications described. In all the above applications, the desired detection limits are met by adopting a broad range of strategies with the focus of keeping the method amenable to use at the point-of-care, i.e., ensuring that sample preparation is completed in <4 minutes. The proof-of-concept results obtained with the custom built Mini-12 mass spectrometer at Purdue University lays the groundwork to (i) encourage the introduction of miniature mass spectrometer-based assays for the molecules listed in a clinical setting and (ii) extend the use of miniature mass spectrometer-based assays for other therapeutic regimens that require longitudinal monitoring.</p>
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Fast and Inexpensive Detection of Bacterial Viability and Drug Susceptibility Through Metabolic MonitoringAyyash, Sondos 15 November 2016 (has links)
Conventional methods for the detection of bacterial infection such as DNA or immunoassays are either expensive, time consuming, or not definitive; thus may not provide all the information sought by the medical professionals. In particular, it is difficult to obtain information about viability or drug effectiveness, which are crucial to formulate a treatment. Bacterial culture test is the “gold standard” because it is inexpensive and does not require extensive sample preparation, and most importantly, provides all the necessary information sought by healthcare professionals, such as bacterial presence, viability and drug effectiveness. These conventional culture methods, however, have a long turnaround time: anywhere between 1 day to 4 weeks. This thesis proposes to solve this problem by monitoring the growth of bacteria in thousands of nanowells simultaneously to identify its presence in the sample and its viability, faster. The segmentation of a sample with low bacterial concentration into thousands of nanoliter wells digitizes the samples and increases the effective concentration in those wells that contain bacteria. The user may then monitor the metabolism of the aerobic bacteria by using an oxygen sensitive fluorophore, ruthenium tris (2,2’-diprydl) dichloride hexahydrate (Ru(Bpy)3) by monitoring the dissolved oxygen concentration in the nanowells. Using E.Coli K12, it is demonstrated that the detection time of E.coli can be as fast as 35-60 minutes with sample concentrations varying from 104(62 minutes for detection), 106 (42 minutes) and 108 cells/mL (38 minutes). More importantly, throughout the thesis it is also demonstrated that reducing the well size can reduce the time of detection. Finally, this thesis will discuss how the drug effectiveness information can be obtained in this format by loading the wells with the drug and monitoring the metabolism of the bacteria. The method that is developed in this thesis is low cost, simple, requires minimal sample preparation and can potentially be used with a wide variety of samples in resource poor setting to detect bacterial infections such as Tuberculosis. / Thesis / Master of Science (MSc)
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Development of analytical techniques for biomedical applications toward point-of-care testing devices / ポイントオブケア検査装置に向けた生物医学的応用のための分析技術の開発Manmana, Yanawut 26 September 2022 (has links)
京都大学 / 新制・課程博士 / 博士(工学) / 甲第24234号 / 工博第5062号 / 新制||工||1790(附属図書館) / 京都大学大学院工学研究科材料化学専攻 / (主査)教授 大塚 浩二, 教授 沼田 圭司, 教授 大内 誠 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DFAM
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Detection Of Sepsis Biomarkers Using MicrofluidicsDamodara, Sreekant January 2021 (has links)
Sepsis is a “life-threatening organ dysfunction caused by a dysregulated host response to infection” that has a widespread impact on human life around the world. It affects more than 1.5 million people, killing at least 250,000 each year in the US alone and affects 90,000 people annually, with estimated mortality rates of up to 30% in Canada. Our understanding of the different biochemical pathways that in the progression of sepsis has improved patient care for sepsis patients. One part of patient care is the use of biomarkers for patient prognosis that draws on the full range of relevant and available information to model the possible outcomes for an individual. Numerous biomarkers have been studied for patient prognosis that includes Procalcitonin (PCT), C-reactive protein (CRP), TNF-α, cfDNA, protein C and PAI 1. Using a panel of multiple biomarkers provided more accuracy in patient prognosis than using individual biomarkers and one such panel that was proposed used cfDNA, protein C, platelet count, creatinine, Glasgow Coma Scale [GCS] score, and lactate. Commercial, low cost POC techniques were available for the measurement of all biomarkers besides cfDNA and protein
C. The objective of this doctoral thesis was chosen to develop low cost, microfluidic devices for the measurement of protein C and cfDNA using nonspecific fluorescence dyes that would enable the eventual integration of the systems and improve patient prognosis. The measurement of protein C in plasma required the separation of protein C from interfering proteins in plasma. This was done through the development of a two-stage separation process that included the development of tunable agarose isoelectric gates for separating proteins using their isoelectric point and the miniaturization of immobilized metal affinity chromatography and its extension to Barium for the selective binding of proteins using their chemical affinity. This was performed in a xurographically fabricated chip to reduce costs and enable the use of geometric focusing of the electric field to enable the operation of the device at a lower applied voltage. The challenges faced with cfDNA were different due to the different characteristics of the material and less interference from plasma. The requirement was to measure the total cfDNA content with minimal cost in comparison to currently available techniques. This was achieved through the development of thread microfluidic devices that showed the use of thread for automated aliquoting of samples by controlling length and twists of the thread. Preconcentration and use of external apparatus was avoided by showing that thread could be used to amplify fluorescence response to a range that was sufficient for the measurement of cfDNA in sepsis patients. A portable fluorescence imaging setup was developed for this purpose and was used in demonstration for the measurement of cfDNA in plasma with sufficient resolution. In conclusion, we developed technologies for rapid and low-cost measurement of protein C and cfDNA using xurographic and thread-based microfluidics that may serve as valuable in improving patient prognosis. / Thesis / Doctor of Philosophy (PhD) / Sepsis is a major reason for hospitalization and cause of death in hospitals worldwide. Its treatment is highly time sensitive with each hour of delay in diagnosis causing a significant increase in chances of death. Due to the wide range of symptoms that can be caused by sepsis, its diagnosis uses a scoring method that relies on the expertise of the onsite doctors and nurses increasing their workload. A more objective system for detection requires the measurement of the quantities of different biomarkers in blood. Biomarkers are proteins present in plasma that change in quantity due to the body’s reaction to sepsis. Several of these biomarkers have been identified and studied for their use in both diagnosing
the presence of sepsis and in predicting the outcome with the current treatment plan. In this PhD study, we chose two of these biomarkers – circulating free DNA (cfDNA) and protein C and developed low-cost techniques for rapidly measuring their concentration in blood plasma. To do this, we made microfluidic devices with techniques that use low-cost materials such as plastic sheets and threads.The device for the measurement of protein C required separating it from many other proteins in plasma. We showed that a device fabricated from stacked plastic sheets and integrated with agarose gels could be used for the measurement of protein C in plasma with sufficient resolution to help with treating septic patients at a cost of less $5 per device. Similarly, we showed that a device that integrated threads with plastic sheets could be used for measuring the quantity of cfDNA in plasma in a portable format within 15 minutes. Overall, we developed tools for rapid measurement of two biomarkers of sepsis using low cost device that cost under $5 to run and could led to improving the quality of care for sepsis patients.
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On-chip Blood Cell/Plasma Separators on Polymer Lab-on-a-Chip for Point-of-Care Clinical DiagnosticsHan, Jungyoup 02 October 2006 (has links)
No description available.
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“Zinc Chip” Reader for Point-Of-Care Quantification of Zinc in Blood SerumSukhavasi, Sowmya 26 September 2011 (has links)
No description available.
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A low-cost and hand-hold PCR microdevice based on water-cooling technologySun, K., Whiteside, Benjamin R., Hebda, Michael J., Fan, Y., Zhang, Y., Xie, Y., Liang, K. 25 September 2023 (has links)
Yes / Polymerase chain reaction (PCR) has become a powerful tool for detecting various diseases due to its high sensitivity and specificity. However, the long thermocycling time and the bulky system have limited the application of PCR devices in Point-of-care testing. Herein, we have proposed an efficient, low-cost, and hand-hold PCR microdevice, mainly including a control module based on water-cooling technology and an amplification module fabricated by 3D printing. The whole device is tiny and can be easily hand-held with a size of about 110 mm × 100 mm × 40 mm and a weight of about 300 g at a low cost of about $170.83. Based on the water-cooling technology, the device can efficiently perform 30 thermal cycles within 46 min at a heating/cooling rate of 4.0/8.1 ℃/s. To test our instrument, plasmid DNA dilutions were amplified with this device; the results demonstrate successful nucleic acid amplification of the …
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Point of care creatinine testing in diagnostic imaging: a feasibility study within the outpatient computed tomography settingSnaith, Beverly, Harris, M.A., Shinkins, B., Messenger, M., Lewington, A., Jordaan, M., Spencer, N. 08 January 2019 (has links)
Yes / Although the risks associated with iodinated contrast administration are acknowledged to be very low, screening of kidney function prior to administration is still standard practice in many hospitals. This study has evaluated the feasibility of implementing a screening form in conjunction with point of care (PoC) creatinine testing as a method to manage the risks of post contrast acute kidney injury (PC-AKI) within the CT imaging pathway.
Method:
Over an eight-week period 300 adult outpatients attending a UK CT department for contrast-enhanced scans were approached. Participants completed a screening questionnaire for co-morbidities linked to kidney dysfunction and consented to have a PoC and laboratory creatinine tests. Comparison was made against with previous baseline blood tests obtained within the preceding 3 months, as required by the study site. Participants were also invited to attend for follow up PoC and laboratory bloods tests at 48–72 h.
Results:
14 patients (4.7%) had a scan-day eGFR below 45mL/min/1.73m2, all identified through screening. The majority of patients (n=281/300; 93.7%) fell in the same risk category based on previous and scan-day blood results. Six PoC test failures were recorded on the scan day. The constant error between the Abbott i-STAT PoC scan-day measurements and the laboratory scan-day measurements was -3.71 (95% CI: -6.41 to -0.50). Five patients had an elevated creatinine (≥25% from baseline) post contrast administration, but no instances of PC-AKI (≥50% from baseline) were identified.
Conclusion:
PoC creatinine testing is a practical method of ensuring renal function and is feasible in the radiology environment. / National Cancer Diagnostics Capacity Fund, NHS England. Andrew Lewington, Bethany Shinkins and Michael Messenger are currently supported by the NIHR MIC- Leeds In Vitro Diagnostics Co-operative (was previously NIHR Leeds Diagnostic Evidence Co-operative)
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Améliorer la prescription médicamenteuse grâce à des mécanismes économiques? / Improving drug prescription thanks to economic mechanismsMichel-Lepage, Audrey 26 February 2016 (has links)
Le mésusage des antibiotiques et l’antibio-résistance sont devenus un cheval de bataille des autorités sanitaires internationales. L’objectif de ma thèse est d’analyser et évaluer les différents types d’incitations qui amèneraient les médecins à « mieux » prescrire des antibiotiques. La première partie est une revue de littérature dans laquelle j’analyse les interactions entre les différents acteurs du système de santé français, les incitations monétaires et les incitations non-monétaires. Les seconde et troisième parties illustrent la première par des analyses principalement empiriques, qui ont été publiées ou envoyées à des revues. La deuxième partie étudie l’utilisation et l’impact d’outils diagnostiques comme aide à la prescription d’antibiotiques avec d’une part les tests de diagnostics rapides pour l’angine, et d’autre part les tests microbiologiques point-of-care, faits « au lit du patient ». La troisième partie analyse le paiement à la performance comme incitation monétaire à l’amélioration des prescriptions médicamenteuses et notamment des antibiotiques. / Antibiotic misuse and antibiotic resistance are increasing concern for international health authorities. The goal of my thesis is to assess the different incentives which would lead physicians to optimize their antibiotic prescriptions. The first part is a literature review analysing interactions between the agents of the French health system. I evaluate also financial and non-monetary incentives. The second and third parts illustrates the first one through empiric analyses, which have been published or sent to a review. The second part studies the use and the impact of diagnostic tests as a help for antibiotics prescribing, with on one hand the use of rapid diagnostic tests detecting tonsillitis, and on the other hand microbiological tests called Point-Of-care made at the patient bedside. The third part analyses the pay-for-performance program as a financial incentive for improving drugs prescriptions and notably antibiotics.
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