Rapid (30-second), equipment-free purification of nucleic acids using easy-to-make dipsticks

Mason, Michael G.; Botella, José Ramón

doi:10.1038/s41596-020-0392-7

Cited by 63 publications

(53 citation statements)

References 36 publications

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“…This creates inherent differences among races and ethnic populations, as well as between healthy individuals and individuals vulnerable to diseases 2 . Therefore, the rapid identification of specific NA has enormous potential in disease diagnosis and management 3 . One of the most common applications of NA detection is molecular-based diagnostic tests or NA testing for the diagnosis of various infectious diseases caused by different microbes and pathogens [4][5][6][7] .…”

Section: Introductionmentioning

confidence: 99%

RNA-extraction-free nano-amplified colorimetric test for point-of-care clinical diagnosis of COVID-19

et al. 2021

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The global pandemic of coronavirus disease 2019 highlights the shortcomings of the current testing paradigm for viral disease diagnostics. Here, we report a stepwise protocol for an RNA-extraction-free nano-amplified colorimetric test for rapid and naked-eye molecular diagnosis of COVID-19. The test employs a unique dual-prong approach that integrates nucleic acid (NA) amplification and plasmonic sensing for point-of-care detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), with a sample-to-assay response time of <1 h. The RNA-extraction-free nanoamplified colorimetric test utilizes plasmonic gold nanoparticles capped with antisense oligonucleotides (ASOs) as a colorimetric reporter to detect the amplified nucleic acid from the COVID-19 causative virus, SARS-CoV-2. The ASOs are specific for the SARS-CoV-2 N-gene, and binding of the ASOs to their target sequence results in the aggregation of the plasmonic gold nanoparticles. This highly specific agglomeration step leads to a change in the plasmonic response of the nanoparticles. Furthermore, when tested using clinical samples, the accuracy, sensitivity and specificity of the test were found to be >98.4%, >96.6% and 100%, respectively, with a detection limit of 10 copies/μL. The test can easily be adapted to diagnose other viral infections with a simple modification of the ASOs and primer sequences. It also provides a low-cost, rapid approach requiring minimal instrumentation that can be used as a screening tool for the diagnosis of COVID-19 at point-of-care settings in resource-poor situations. The colorimetric readout of the test can even be monitored using a handheld optical reader to obtain a quantitative response. Therefore, we anticipate that this protocol will be widely useful for the development of biosensors for the molecular diagnostics of COVID-19 and other infectious diseases.

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Section: Introductionmentioning

confidence: 99%

RNA-extraction-free nano-amplified colorimetric test for point-of-care clinical diagnosis of COVID-19

et al. 2021

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“…However, it should be noted that the risk of cross-contamination of samples on FTA cards is higher compared with liquid samples collected in separate tubes. A further disadvantage of the FTA cards is the less efficient nucleic acid extraction based on the reduced recovery of the DNA and RNA from the filter paper matrix [ 11 ]. Although the probability of detecting the pathogens on FTA cards is lower than in fresh samples, the cards offer unique advantages for the collection, transportation, and storage of samples.…”

Section: Introductionmentioning

confidence: 99%

Optimizing Release of Nucleic Acids of African Swine Fever Virus and Influenza A Virus from FTA Cards

Elnagar

Harder

Blome

et al. 2021

IJMS

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FTA cards and related products simplify the collection, transport, and transient storage of biological sample fluids. Here, we have compared the yield and quality of DNA and RNA released from seven different FTA cards using seven releasing/extraction methods with eleven experimental eluates. For the validation, dilution series of African swine fever virus (ASFV) positive EDTA blood and Influenza A virus (IAV) positive allantoic fluid were used. Based on our data, we conclude that direct PCR amplification without the need for additional nucleic acid extraction and purification could be suitable and more convenient for ASFV DNA release from FTA cards. In contrast, IAV RNA loads can be amplified from FTA card punches if a standard extraction procedure including a lysis step is applied. These differences between the amplifiable viral DNA and RNA after releasing and extraction are not influenced by the type of commercial FTA card or the eleven different nucleic acid releasing procedures used for the comparative analyses. In general, different commercial FTA cards were successfully used for the storage and recovery of the ASFV and IAV genetic material suitable for PCR. Nevertheless, the usage of optimized nucleic acid releasing protocols could improve the recovery of the viral genome of both viruses. Here, the application of Chelex® Resin 100 buffer mixed with 1 × Tris EDTA buffer (TE, pH 8.0) or with TED 10 (TE buffer and Dimethylsulfoxid) delivered the best results and can be used as a universal method for releasing viral DNA and RNA from FTA cards.

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“…Therefore, our analysis highlights the importance of empirically validating these components as main priorities in an assay establishment. tions (Mason & Botella, 2020;Zou et al, 2017). This rapid method can extract nucleic acids from plants, microbes and animals within 30 s while performing comparably to conventional techniques and not requiring pipetting (Sullivan et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

CRISPR‐based platform for rapid, sensitive and field‐deployable detection of scale drop disease virus in Asian sea bass (Lates calcarifer)

Sukonta

Senapin

Meemetta

et al. 2021

Journal of Fish Diseases

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Asian sea bass or barramundi (Lates calcarifer) is one of the most important farmed fish species in South-East Asia and Australia, with the global production steadily rising in the past decade and reaching 203,064 tonnes in 2019 (FAO, 2021;Nurliyana et al., 2020).However, the high demand for sea bass has spurred rapid expansion and intensification of cultivation that in turn exacerbate the risk of disease outbreaks (Hutson, 2013). A major viral disease of Asian sea bass is scale drop disease (SDD), caused by scale drop disease

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Rapid (30-second), equipment-free purification of nucleic acids using easy-to-make dipsticks

Cited by 63 publications

References 36 publications

RNA-extraction-free nano-amplified colorimetric test for point-of-care clinical diagnosis of COVID-19

RNA-extraction-free nano-amplified colorimetric test for point-of-care clinical diagnosis of COVID-19

Optimizing Release of Nucleic Acids of African Swine Fever Virus and Influenza A Virus from FTA Cards

CRISPR‐based platform for rapid, sensitive and field‐deployable detection of scale drop disease virus in Asian sea bass (Lates calcarifer)

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