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  1. Article ; Online: Sample-to-Answer Immuno-Magnetic Assay Using Thermally Responsive Alkane Partitions.

    Everitt, Micaela L / Boegner, David J / White, Ian M

    Biosensors

    2022  Volume 12, Issue 11

    Abstract: To combat pandemics, there is a need for rapid point-of-care diagnostics to identify infected patients and to track the spread of the disease. While recent progress has been made in response to COVID-19, there continues to be a need for point-of-care ... ...

    Abstract To combat pandemics, there is a need for rapid point-of-care diagnostics to identify infected patients and to track the spread of the disease. While recent progress has been made in response to COVID-19, there continues to be a need for point-of-care diagnostics capable of detecting biomarkers-such as antibodies-in whole blood. We have recently reported the development of thermally responsive alkane partitions (TRAPs) for the automation of point-of-care immuno-magnetic assays. Here, we demonstrate the use of TRAPs to enable sample-to-answer detection of antibodies against the SARS-CoV-2 virus in whole blood samples. We report a limit of detection of 84 pg/mL, well below the clinically relevant threshold. We anticipate that the TRAP-enabled sample-to-answer immunoassay can be used to track the progression of future pandemics, leading to a more informed and robust clinical and societal response.
    MeSH term(s) Humans ; Alkanes ; SARS-CoV-2 ; COVID-19/diagnosis ; Biological Assay ; Antibodies, Viral
    Chemical Substances Alkanes ; Antibodies, Viral
    Language English
    Publishing date 2022-11-17
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2662125-3
    ISSN 2079-6374 ; 2079-6374
    ISSN (online) 2079-6374
    ISSN 2079-6374
    DOI 10.3390/bios12111030
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Thermally Responsive Alkane Partitions for Assay Automation.

    Boegner, David J / Everitt, Micaela L / White, Ian M

    ACS applied materials & interfaces

    2022  Volume 14, Issue 7, Page(s) 8865–8875

    Abstract: For point-of-care diagnostic tools to be impactful, they must be inexpensive, equipment-free, and sample-to-answer (i.e., require no user intervention). Here, we report a new approach to enable sample-to-answer diagnostics that utilizes thermally ... ...

    Abstract For point-of-care diagnostic tools to be impactful, they must be inexpensive, equipment-free, and sample-to-answer (i.e., require no user intervention). Here, we report a new approach to enable sample-to-answer diagnostics that utilizes thermally responsive alkane partitions (TRAPs) as automated pseudo-valves. When combined with the magnetic manipulation of microbeads, TRAPs enable the pumpless automation of all steps in complex assays. We demonstrate that in relatively narrow channel geometries, liquified alkane partitions continue to separate reagents on each side of the partition while enabling the transition of magnetic beads from one reagent to the next, replacing manual pipetting steps in conventional assays. In addition, we show that in relatively broader geometries, liquified partitions breach, enabling the addition/mixing of preloaded reagents. Through calculation and experimentation, we determine the geometric design rules for implementing the stationary and removable partitions in fluidic channels. In addition, we demonstrate that magnetic microbeads can be pulled through liquified stationary TRAPs without disrupting partition integrity and without disrupting bound protein complexes attached at the microbead surface. The TRAP technology introduced here can enable a new low-cost and equipment-free approach for fully automated sample-to-answer diagnostics.
    MeSH term(s) Alkanes ; Automation ; Biological Assay ; Magnetics ; Point-of-Care Systems
    Chemical Substances Alkanes
    Language English
    Publishing date 2022-02-11
    Publishing country United States
    Document type Journal Article
    ISSN 1944-8252
    ISSN (online) 1944-8252
    DOI 10.1021/acsami.2c00493
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Sample-to-Answer Diagnostic System for the Detection of Circulating Histones in Whole Blood.

    Everitt, Micaela L / Boegner, David J / Birukov, Konstantin G / White, Ian M

    ACS sensors

    2021  Volume 6, Issue 8, Page(s) 3006–3012

    Abstract: Severe internal trauma results in millions of hospitalizations each year, including thousands of deaths caused by subsequent multiple organ failure. The majority of these deaths occur within the first 24 h, and thus, rapid diagnosis of internal trauma ... ...

    Abstract Severe internal trauma results in millions of hospitalizations each year, including thousands of deaths caused by subsequent multiple organ failure. The majority of these deaths occur within the first 24 h, and thus, rapid diagnosis of internal trauma severity is necessary for immediate treatment. For early organ damage identification, diagnosis in point-of-care settings is crucial for rapid triage and treatment. Recent reports suggest that circulating histones may serve as a biomarker for severe organ damage and the risk of multiple organ failure. Here, we report a point-of-care diagnostic system that utilizes the inherent interactions between histones and DNA for the fluorescence-based detection of histones in whole blood. In the assay, histones within the sample are wrapped by DNA, thus preventing an intercalating dye from binding the DNA and fluorescing. To allow for quantitative fluorescent measurements to be made in a point-of-care setting, we integrate a rapid, automated blood separation step into our assay. Furthermore, we eliminate manual reagent additions using a thermally responsive alkane partition (TRAP), thus making the system sample-to-answer. Finally, we demonstrate the assay in a portable fluorescence reader compatible with a point-of-care environment. We report a limit of detection 112 ng/mL in whole blood, suggesting that our device can be used to rapidly diagnose internal trauma severity and the likelihood of multiple organ failure in near-patient settings.
    MeSH term(s) Biomarkers ; Histones ; Humans ; Point-of-Care Systems
    Chemical Substances Biomarkers ; Histones
    Language English
    Publishing date 2021-07-16
    Publishing country United States
    Document type Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
    ISSN 2379-3694
    ISSN (online) 2379-3694
    DOI 10.1021/acssensors.1c00886
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article: Thermally Responsive Alkane Partitions for Assay Automation

    Boegner, David J. / Everitt, Micaela L. / White, Ian M.

    ACS applied materials & interfaces. 2022 Feb. 11, v. 14, no. 7

    2022  

    Abstract: For point-of-care diagnostic tools to be impactful, they must be inexpensive, equipment-free, and sample-to-answer (i.e., require no user intervention). Here, we report a new approach to enable sample-to-answer diagnostics that utilizes thermally ... ...

    Abstract For point-of-care diagnostic tools to be impactful, they must be inexpensive, equipment-free, and sample-to-answer (i.e., require no user intervention). Here, we report a new approach to enable sample-to-answer diagnostics that utilizes thermally responsive alkane partitions (TRAPs) as automated pseudo-valves. When combined with the magnetic manipulation of microbeads, TRAPs enable the pumpless automation of all steps in complex assays. We demonstrate that in relatively narrow channel geometries, liquified alkane partitions continue to separate reagents on each side of the partition while enabling the transition of magnetic beads from one reagent to the next, replacing manual pipetting steps in conventional assays. In addition, we show that in relatively broader geometries, liquified partitions breach, enabling the addition/mixing of preloaded reagents. Through calculation and experimentation, we determine the geometric design rules for implementing the stationary and removable partitions in fluidic channels. In addition, we demonstrate that magnetic microbeads can be pulled through liquified stationary TRAPs without disrupting partition integrity and without disrupting bound protein complexes attached at the microbead surface. The TRAP technology introduced here can enable a new low-cost and equipment-free approach for fully automated sample-to-answer diagnostics.
    Keywords alkanes ; automation ; diagnostic techniques ; geometry ; magnetism ; microbeads ; point-of-care systems
    Language English
    Dates of publication 2022-0211
    Size p. 8865-8875.
    Publishing place American Chemical Society
    Document type Article
    ISSN 1944-8252
    DOI 10.1021/acsami.2c00493
    Database NAL-Catalogue (AGRICOLA)

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  5. Article ; Online: A critical review of point-of-care diagnostic technologies to combat viral pandemics.

    Everitt, Micaela L / Tillery, Alana / David, Martha G / Singh, Nikita / Borison, Aviva / White, Ian M

    Analytica chimica acta

    2020  Volume 1146, Page(s) 184–199

    Abstract: The COVID-19 global pandemic of 2019-2020 pointedly revealed the lack of diagnostic solutions that are able to keep pace with the rapid spread of the virus. Despite the promise of decades of lab-on-a-chip research, no commercial products were available ... ...

    Abstract The COVID-19 global pandemic of 2019-2020 pointedly revealed the lack of diagnostic solutions that are able to keep pace with the rapid spread of the virus. Despite the promise of decades of lab-on-a-chip research, no commercial products were available to deliver rapid results or enable testing in the field at the onset of the pandemic. In this critical review, we assess the current state of progress on the development of point-of-care technologies for the diagnosis of viral diseases that cause pandemics. While many previous reviews have reported on progress in various lab-on-a-chip technologies, here we address the literature from the perspective of the testing needs of a rapidly expanding pandemic. First, we recommend a set of requirements to heed when designing point-of-care diagnostic technologies to address the testing needs of a pandemic. We then review the current state of assay technologies with a focus on isothermal amplification and lateral-flow immunoassays. Though there is much progress on assay development, we argue that the largest roadblock to deployment exists in sample preparation. We summarize current approaches to automate sample preparation and discuss both the progress and shortcomings of these developments. Finally, we provide our recommendations to the field of specific challenges to address in order to prepare for the next pandemic.
    MeSH term(s) COVID-19/diagnosis ; Humans ; Lab-On-A-Chip Devices ; Pandemics ; Point-of-Care Systems/trends ; Point-of-Care Testing/trends
    Language English
    Publishing date 2020-10-11
    Publishing country Netherlands
    Document type Journal Article ; Review
    ZDB-ID 1483436-4
    ISSN 1873-4324 ; 0003-2670
    ISSN (online) 1873-4324
    ISSN 0003-2670
    DOI 10.1016/j.aca.2020.10.009
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Phenotypically distinguishing ESBL-producing pathogens using paper-based surface enhanced Raman sensors.

    Hilton, Shannon H / Hall, Connor / Nguyen, Hieu T / Everitt, Micaela L / DeShong, Philip / White, Ian M

    Analytica chimica acta

    2020  Volume 1127, Page(s) 207–216

    Abstract: Antimicrobial stewardship practices are critical in preventing the further erosion of treatment options for bacterial infections. Yet, at the same time, determination of an infection's antimicrobial susceptibility requires multiple rounds of culture and ... ...

    Abstract Antimicrobial stewardship practices are critical in preventing the further erosion of treatment options for bacterial infections. Yet, at the same time, determination of an infection's antimicrobial susceptibility requires multiple rounds of culture and expensive lab automation systems. In this work, we report the use of paper-based surface enhanced Raman spectroscopy (SERS) sensors and portable instrumentation to phenotypically discriminate multi-drug resistance with fewer culture steps than conventional clinical microbiology. Specifically, we demonstrate the identification of resistance to varying generations of β-lactam antibiotics by detecting the activity of particular β-lactamase enzymes in a multiplexed assay. The method utilizes molecular reporters that consist of β-lactams with SERS barcodes. Hydrolysis of the β-lactam by β-lactamase enzymes in the sample expels the barcode; the released sulfur-containing barcode is then detected via SERS. Using this approach, we demonstrate the differentiation of E. coli strains with (1) extended spectrum β-lactamase (ESBL), (2) narrow-spectrum β-lactamase, and (3) no resistance, using only a single measurement on a single sample. In addition, we experimentally validate an approach to expand the library of reporters through the simple chemical synthesis of new barcoded β-lactams. Importantly, the reported method determines the susceptibility based on phenotypic β-lactamase activity, which is aligned with current microbiology lab standards. This new method will enable the precise selection of effective β-lactam antibiotics (as opposed to defaulting to drugs of last resort) faster than current methods while using simple steps and low-cost portable instrumentation.
    MeSH term(s) Anti-Bacterial Agents/pharmacology ; Escherichia coli ; Escherichia coli Infections ; Humans ; Microbial Sensitivity Tests ; beta-Lactamases
    Chemical Substances Anti-Bacterial Agents ; beta-Lactamases (EC 3.5.2.6)
    Language English
    Publishing date 2020-07-11
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 1483436-4
    ISSN 1873-4324 ; 0003-2670
    ISSN (online) 1873-4324
    ISSN 0003-2670
    DOI 10.1016/j.aca.2020.06.068
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: A critical review of point-of-care diagnostic technologies to combat viral pandemics

    Everitt, Micaela L. / Tillery, Alana / David, Martha G. / Singh, Nikita / Borison, Aviva / White, Ian M.

    Anal Chim Acta

    Abstract: The COVID-19 global pandemic of 2019-2020 pointedly revealed the lack of diagnostic solutions that are able to keep pace with the rapid spread of the virus. Despite the promise of decades of lab-on-a-chip research, no commercial products were available ... ...

    Abstract The COVID-19 global pandemic of 2019-2020 pointedly revealed the lack of diagnostic solutions that are able to keep pace with the rapid spread of the virus. Despite the promise of decades of lab-on-a-chip research, no commercial products were available to deliver rapid results or enable testing in the field at the onset of the pandemic. In this critical review, we assess the current state of progress on the development of point-of-care technologies for the diagnosis of viral diseases that cause pandemics. While many previous reviews have reported on progress in various lab-on-a-chip technologies, here we address the literature from the perspective of the testing needs of a rapidly expanding pandemic. First, we recommend a set of requirements to heed when designing point-of-care diagnostic technologies to address the testing needs of a pandemic. We then review the current state of assay technologies with a focus on isothermal amplification and lateral-flow immunoassays. Though there is much progress on assay development, we argue that the largest roadblock to deployment exists in sample preparation. We summarize current approaches to automate sample preparation and discuss both the progress and shortcomings of these developments. Finally, we provide our recommendations to the field of specific challenges to address in order to prepare for the next pandemic.
    Keywords covid19
    Publisher Elsevier; PMC
    Document type Article ; Online
    DOI 10.1016/j.aca.2020.10.009
    Database COVID19

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