Molecular Diagnostic Field Test for Point-of-Care Detection of Ebola Virus Directly From Blood

Benzine, Jason W.; Brown, Kerry M.; Agans, Krystle N.; Godiska, Ronald; Mire, Chad E.; Gowda, Krishne; Converse, Brandon J.; Geisbert, Thomas W.; Mead, David A.; Chander, Yogesh

doi:10.1093/infdis/jiw330

Cited by 32 publications

(28 citation statements)

References 21 publications

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“…Moreover, both systems are ‘closed access’, meaning public health authorities cannot independently respond to emerging threats by rapidly transitioning their in-house developed assays to such disseminated use platforms: tellingly, the Trombley assay received FDA EUA days after the WHO PHEIC declaration, whereas the Cepheid platform required several months of additional development. Elsewhere, emerging alternative NAAT solutions focus on the energy efficiency of isothermal amplification technologies 64 as well as the agnostic, de novo analytical power of memory stick-sized sequencing systems. 65 , 66 Whilst many of these have been successfully evaluated in resource-limited settings, 64 , 65 these solutions still require extensive and costly laboratory infrastructure for sample pre-processing 64 – 66 even onto genome extraction, 65 , 66 are non-quantitative, 64 closed access, 64 , 66 or require high bandwidth data transfer/computational power access 65 , 66 to realize their potential.…”

Section: Discussionmentioning

confidence: 99%

Field-deployable, quantitative, rapid identification of active Ebola virus infection in unprocessed blood

et al. 2017

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Section: Discussionmentioning

confidence: 99%

Field-deployable, quantitative, rapid identification of active Ebola virus infection in unprocessed blood

et al. 2017

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“…Using the 10× on-chip concentration protocol, our technique shows a detection limit of 800 aM for Ebola RNA targets, which is comparable with PCR analysis. 51 This high sensitivity can provide crucial information for early detection by confirming or clearing suspected cases. 52 Thus, our technique is compatible with the fingerprick test and is easy to carry out in resource-limited settings.…”

Section: Discussionmentioning

confidence: 99%

Microfluidic System for Detection of Viral RNA in Blood Using a Barcode Fluorescence Reporter and a Photocleavable Capture Probe

Park

Griffiths

et al. 2017

Anal. Chem.

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A microfluidic sample preparation multiplexer (SPM) and assay procedure is developed to improve amplification-free detection of Ebola virus RNA from blood. While a previous prototype successfully detected viral RNA following off-chip RNA extraction from infected cells, the new device and protocol can detect Ebola virus in raw blood with clinically relevant sensitivity. The Ebola RNA is hybridized with sequence specific capture and labeling DNA probes in solution and then the complex is pulled down onto capture beads for purification and concentration. After washing, the captured RNA target is released by irradiating the photocleavable DNA capture probe with ultraviolet (UV) light. The released, labeled, and purified RNA is detected by a sensitive and compact fluorometer. Exploiting these capabilities, a detection limit of 800 attomolar (aM) is achieved without target amplification. The new SPM can run up to 80 assays in parallel using a pneumatic multiplexing architecture. Importantly, our new protocol does not require time-consuming and problematic off-chip probe conjugation and washing. This improved SPM and labeling protocol is an important step toward a useful POC device and assay.

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“…Three million people are estimated to be at risk of dengue fever 82 , caused by Dengue virus that is transmitted by mosquitos and widespread in Latin America and Asia 83 . Dengue fever symptoms resemble those of hemorrhagic fevers caused by Ebola virus and other viruses 84 . Thus, laboratory diagnosis of dengue fever is crucial for rapid management of patients with severe infection 85 .…”

Section: Crispr-tools For Emerging and Re-emerging Infectious Diseasesmentioning

confidence: 96%

CRISPR-Cas Systems for Diagnosing Infectious Diseases

Kostyusheva¹,

Brezgin²,

Babin³

et al. 2020

Preprint

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Infectious diseases are a global health problem affecting billions of people. Developing rapid and sensitive diagnostic tools is key for successful patient management and curbing disease spread. Currently available diagnostics are very specific and sensitive but time-consuming and require expensive laboratory settings and well-trained personnel; thus, they are not available in resource-limited areas, for the purposes of large-scale screenings and in case of outbreaks and epidemics. Developing new, rapid, and affordable point-of-care diagnostic assays is urgently needed. This review focuses on CRISPR-based technologies and their perspectives to become platforms for point-of-care nucleic acid detection methods and as deployable diagnostic platforms that could help to identify and curb outbreaks and emerging epidemics. We describe the mechanisms and function of different classes and types of CRISPR-Cas systems, including pros and cons for developing molecular diagnostic tests and applications of each type to detect a wide range of infectious agents. Many Cas proteins (Cas9, Cas12, Cas13, Cas14) have been leveraged to create highly accurate and sensitive diagnostic tools combined with technologies of signal amplification and fluorescent, potentiometric, colorimetric, or lateral flow assay detection. In particular, the most advanced platforms -- SHERLOCK/v2, DETECTR, or CRISPR-Chip -- enable detection of attomolar amounts of pathogenic nucleic acids with specificity comparable to that of PCR but with minimal technical settings. Further developing CRISPR-based diagnostic tools promises to dramatically transform molecular diagnostics, making them easily affordable and accessible virtually anywhere in the world. The burden of socially significant diseases, frequent outbreaks, recent epidemics (MERS, SARS and the ongoing coronoviral nCov-2019 infection) urgently need the developing of express-diagnostic tools. Recently devised CRISPR-technologies represent the unprecedented opportunity to reshape epidemiological surveillance and molecular diagnostics.

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Molecular Diagnostic Field Test for Point-of-Care Detection of Ebola Virus Directly From Blood

Cited by 32 publications

References 21 publications

Field-deployable, quantitative, rapid identification of active Ebola virus infection in unprocessed blood

Field-deployable, quantitative, rapid identification of active Ebola virus infection in unprocessed blood

Microfluidic System for Detection of Viral RNA in Blood Using a Barcode Fluorescence Reporter and a Photocleavable Capture Probe

CRISPR-Cas Systems for Diagnosing Infectious Diseases

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