SHERLOCK and DETECTR: CRISPR-Cas Systems as Potential Rapid Diagnostic Tools for Emerging Infectious Diseases

Mustafa, Mujahed I.; Makhawi, Abdelrafie M.

doi:10.1128/jcm.00745-20

Cited by 165 publications

(114 citation statements)

References 73 publications

(117 reference statements)

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“…Cas13 and Cas12a are activated upon binding to target nucleic acids, RNA and DNA, respectively, where they excise reporter RNA sequences and cut a quenched fluorescent probe to generate a fluorescence signal (187)(188)(189). Both tests are low-cost, can be performed in 1 h (188,190,191), and have been granted U.S. FDA EUA status (72,192,193). The SHERLOCK test demonstrated a sensitivity of 93.1% and a specificity of 98.5% (191), while the DETECTR assay demonstrated 95% positive predictive agreement and 100% negative predictive agreement (188), which makes both strong rapid diagnostic candidates.…”

Section: Other Methods Of Viral Rna Detectionmentioning

confidence: 99%

Review of Current COVID-19 Diagnostics and Opportunities for Further Development

Mardian¹,

Kosasih²,

et al. 2021

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Diagnostic testing plays a critical role in addressing the coronavirus disease 2019 (COVID-19) pandemic, caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Rapid and accurate diagnostic tests are imperative for identifying and managing infected individuals, contact tracing, epidemiologic characterization, and public health decision making. Laboratory testing may be performed based on symptomatic presentation or for screening of asymptomatic people. Confirmation of SARS-CoV-2 infection is typically by nucleic acid amplification tests (NAAT), which requires specialized equipment and training and may be particularly challenging in resource-limited settings. NAAT may give false-negative results due to timing of sample collection relative to infection, improper sampling of respiratory specimens, inadequate preservation of samples, and technical limitations; false-positives may occur due to technical errors, particularly contamination during the manual real-time polymerase chain reaction (RT-PCR) process. Thus, clinical presentation, contact history and contemporary phyloepidemiology must be considered when interpreting results. Several sample-to-answer platforms, including high-throughput systems and Point of Care (PoC) assays, have been developed to increase testing capacity and decrease technical errors. Alternatives to RT-PCR assay, such as other RNA detection methods and antigen tests may be appropriate for certain situations, such as resource-limited settings. While sequencing is important to monitor on-going evolution of the SARS-CoV-2 genome, antibody assays are useful for epidemiologic purposes. The ever-expanding assortment of tests, with varying clinical utility, performance requirements, and limitations, merits comparative evaluation. We herein provide a comprehensive review of currently available COVID-19 diagnostics, exploring their pros and cons as well as appropriate indications. Strategies to further optimize safety, speed, and ease of SARS-CoV-2 testing without compromising accuracy are suggested. Access to scalable diagnostic tools and continued technologic advances, including machine learning and smartphone integration, will facilitate control of the current pandemic as well as preparedness for the next one.

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Section: Other Methods Of Viral Rna Detectionmentioning

confidence: 99%

Review of Current COVID-19 Diagnostics and Opportunities for Further Development

Mardian¹,

Kosasih²,

et al. 2021

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“…However, studies have shown specificity between 71.4% and 98.5% for CRISPR, 78 , 79 so further evaluations are required since a recent review has concluded that although CRISPR technology is still in its infancy it is potentially revolutionary as tests are rapid and portable. 80 …”

Section: Other Novel Emerging Technologiesmentioning

confidence: 99%

Use of emerging testing technologies and approaches for SARS-CoV-2: review of literature and global experience in an Australian context

et al. 2021

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Emerging testing technologies for detection of SARS-CoV-2 include those that are rapid and can be used at point-of-care (POC), and those facilitating high throughput laboratory-based testing. Tests designed to be performed at POC (such as antigen tests and molecular assays) have the potential to expedite isolation of infectious patients and their contacts, but most are less sensitive than standard-of-care reverse transcription polymerase chain reaction (RT-PCR). Data on clinical performance of the majority of emerging assays is limited with most evaluations performed on contrived or stored laboratory samples. Further evaluations of these assays are required, particularly when performed at POC on symptomatic and asymptomatic patients and at various time-points after symptom onset. A few studies have so far shown several of these assays have high specificity. However, large prospective evaluations are needed to confirm specificity, particularly before the assays are implemented in low prevalence settings or asymptomatic populations. High throughput laboratory-based testing includes the use of new sample types (e.g., saliva to increase acceptability) or innovative uses of existing technology (e.g., sample pooling). Information detailing population-wide testing strategies for SARS-COV-2 is largely missing from peer-reviewed literature. Logistics and supply chains are key considerations in any plan to ‘scale up’ testing in the Australian context. The strategic use of novel assays will help strike the balance between achieving adequate test numbers without overwhelming laboratory capacity. To protect testing of high-risk populations, the aims of testing with respect to the phase of the pandemic must be considered.

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“…A growing number of preclinical studies based on rodent and other animal models indicate that CRISPR-Cas systems have the potential for therapeutic usage in different diseases, including genetic diseases, 31 infectious diseases, 32 cancers, 7 , 33 immunological diseases (autoimmunity and immunodeficiency), 34 etc.…”

Section: Preclinical Testsmentioning

confidence: 99%

Applications and challenges of CRISPR-Cas gene-editing to disease treatment in clinics

Liu

Jiang

et al. 2021

Precision Clinical Medicine

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Clustered regularly interspaced short palindromic repeats-CRISPR associated systems (Cas) are efficient tools for targeting specific genes for laboratory research, agricultural engineering, biotechnology, and human disease treatment. Cas9, by far the most extensively used gene-editing nuclease, has shown great promise for the treatment of hereditary diseases, viral infection, cancers and so on. Recent reports have revealed that some other types of CRISPR-Cas systems may also have surprising potential to join the fray as gene-editing tools for various applications. Despite the fast progress in basic researches and clinical tests, some underlying problems present continuous, significant challenges, such as editing efficiency, relative difficulty in delivery, off-target effects, immunogenicity, etc. This article summarizes the applications of CRISPR-Cas from bench to bedside and highlights the current obstacles that may limit the usage of CRISPR-Cas systems as gene-editing tool-kits in precision medicine and offer some viewpoints that may help to tackle these challenges and facilitate technical development. CRISPR-Cas systems, as a powerful gene-editing approach, will offer great hopes in clinical treatments for many individuals with currently incurable diseases.

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SHERLOCK and DETECTR: CRISPR-Cas Systems as Potential Rapid Diagnostic Tools for Emerging Infectious Diseases

Cited by 165 publications

References 73 publications

Review of Current COVID-19 Diagnostics and Opportunities for Further Development

Review of Current COVID-19 Diagnostics and Opportunities for Further Development

Use of emerging testing technologies and approaches for SARS-CoV-2: review of literature and global experience in an Australian context

Applications and challenges of CRISPR-Cas gene-editing to disease treatment in clinics

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