Nucleic acid extraction from complex biofluid using toothpick-actuated over-the-counter medical-grade cotton

Kumar, Shrawan; Kharb, Anjali; Vazirani, Aman; Chauhan, Rajinder Singh; Pramanik, Goutam; Sengupta, Mrittika; Ghosh, Souradyuti

doi:10.1016/j.bmc.2022.117009

Cited by 4 publications

(4 citation statements)

References 41 publications

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“…The utility of RAA‐LF system is, however, dependent on the quality of nucleic acid templates to be detected, and the amplification‐inhibiting factors may affect assay's efficacy as well as accuracy 28 . Sample preparation is often a multistep procedure requiring columns or magnetic beads and other laboratory instruments.…”

Section: Resultsmentioning

confidence: 99%

“…The utility of RAA-LF system is, however, dependent on the quality of nucleic acid templates to be detected, and the amplificationinhibiting factors may affect assay's efficacy as well as accuracy. 28 Sample preparation is often a multistep procedure requiring columns or magnetic beads and other laboratory instruments. To develop an one-step sample preparation method suitable for the RAA-LF assay, we infected mice with VACV via the nasal route for 7 days, used swabs to collect samples from different sites, and directly soaked the swabs in 1× sample lysis buffer.…”

Section: One-step Sample Dna Preparation and Raa-lf Detectionmentioning

confidence: 99%

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A field diagnostic method for rapid and sensitive detection of mpox virus

Zhao,

Xu,

Wang

et al. 2024

Journal of Medical Virology

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The mpox outbreak has subdued with fewer reported cases at the present in high‐income countries. It is known that mpox virus (MPXV) infection has been epidemic for more than 50 years in African countries. The ancestral MPXV strain has changed into multiple clades, indicating the ongoing evolution of MPXV, which reflects the historical neglect of mpox in Africa, especially after smallpox eradication, and bestows the danger of more severe mpox epidemics in the future. It is thus imperative to continue the development of mpox diagnostics and treatments so we can be prepared in the event of a new mpox epidemic. In this study, we have developed an MPXV detection tool that leverages the recombinase‐aid amplification assay by integrating lateral flow strips (RAA‐LF) and one‐step sample DNA preparation, with visible readout, no need of laboratory instrument, and ready for field deployment. The detection limit reaches 10 copies per reaction. The performance of our RAA‐FL assay in diagnosing mpox clinical samples is on par with that of the quantitative polymerase chain reaction (PCR) assay. Taken together, we have developed a point‐of‐care RAA‐LF method of high accuracy and sensitivity, readily deployable for field detection of MPXV. This diagnostic tool is expected to improve and accelerate field‐ and self‐diagnosis, allow timely isolation and treatment, reduce the spread of MPXV, thus effectively mitigate MPXV outbreak in the future.

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Section: Resultsmentioning

confidence: 99%

Section: One-step Sample Dna Preparation and Raa-lf Detectionmentioning

confidence: 99%

A field diagnostic method for rapid and sensitive detection of mpox virus

Zhao,

Xu,

Wang

et al. 2024

Journal of Medical Virology

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“… 47 , 48 In addition, point-of-care or field-deployable MPXV diagnostics will require rapid and easy sample preparation. A few methods of sample preparations have been reported for point-of care or field-deployable diagnostics, including one-step sample extraction with TNA-Cifer Reagent, 49 using dipstick extraction technology, 50 and disposable pen-like sensor systems, 51 which can be tested along with our mpox detection assay. Our RPA-Cas12a system can be optimized by incorporating the visual readout component and one-step sample preparations with the aim for field and home use.…”

Section: Discussionmentioning

confidence: 99%

Rapid and sensitive one-tube detection of mpox virus using RPA-coupled CRISPR-Cas12 assay

Zhao,

Hu,

Fan

et al. 2023

Cell Reports Methods

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“…The risk of the inhibitory effect of chaotropic reagents has made SPE methods that are not reliant on chaotropes an attractive alternative, and several approaches have been reported to eliminate the probability of chemical contamination with the reagents [31,129]. Non-chaotropic approaches to facilitate DNA isolation have used crowding or crosslinking reagents in the NA-SPE process (Figure 3).…”

Section: Non-chaotropic Bindingmentioning

confidence: 99%

Chemical Trends in Sample Preparation for Nucleic Acid Amplification Testing (NAAT): A Review

Lee,

Balakrishnan,

Doeven

et al. 2023

Biosensors

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Nucleic acid amplification testing facilitates the detection of disease through specific genomic sequences and is attractive for point-of-need testing (PONT); in particular, the early detection of microorganisms can alert early response systems to protect the public and ecosystems from widespread outbreaks of biological threats, including infectious diseases. Prior to nucleic acid amplification and detection, extensive sample preparation techniques are required to free nucleic acids and extract them from the sample matrix. Sample preparation is critical to maximize the sensitivity and reliability of testing. As the enzymatic amplification reactions can be sensitive to inhibitors from the sample, as well as from chemicals used for lysis and extraction, avoiding inhibition is a significant challenge, particularly when minimising liquid handling steps is also desirable for the translation of the assay to a portable format for PONT. The reagents used in sample preparation for nucleic acid testing, covering lysis and NA extraction (binding, washing, and elution), are reviewed with a focus on their suitability for use in PONT.

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Nucleic acid extraction from complex biofluid using toothpick-actuated over-the-counter medical-grade cotton

Cited by 4 publications

References 41 publications

A field diagnostic method for rapid and sensitive detection of mpox virus

A field diagnostic method for rapid and sensitive detection of mpox virus

Rapid and sensitive one-tube detection of mpox virus using RPA-coupled CRISPR-Cas12 assay

Chemical Trends in Sample Preparation for Nucleic Acid Amplification Testing (NAAT): A Review

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