Homebrew: An economical and sensitive glassmilk-based nucleic-acid extraction method for SARS-CoV-2 diagnostics

Page, Robert L.; Scourfield, Edward J.; Ficarelli, Mattia; McKellar, Stuart; Lee, Kwok Leung; Maguire, Thomas J. A.; Bouton, Clément R.; Lista, María José; Neil, Stuart J. D.; Malim, Michael H.; Zuckerman, Mark; Mischo, Hannah E.; Martínez-Nuñez, Rocío T.

doi:10.1016/j.crmeth.2022.100186

Cited by 7 publications

(9 citation statements)

References 23 publications

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“…The “glass milk” protocol was based on the one described by Page et al ( Page et al, 2022a , 2022b ) with slight modifications. The pellet was resuspended in 100 μL of nuclease-free water and 100 μL of a 4 % SDS solution.…”

Section: Methodsmentioning

confidence: 99%

Moving towards on-site detection of Shiga toxin-producing Escherichia coli in ready-to-eat leafy greens

Costa-Ribeiro,

Lamas,

Mora

et al. 2024

Current Research in Food Science

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Section: Methodsmentioning

confidence: 99%

Moving towards on-site detection of Shiga toxin-producing Escherichia coli in ready-to-eat leafy greens

Costa-Ribeiro,

Lamas,

Mora

et al. 2024

Current Research in Food Science

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“…We developed our in-house extraction method for nasal swabs called homebrew 11 . We further developed this method for RNA extraction from saliva and compared its performance in 980 samples (Figure 3A and Supplemental Table 3), with homebrew slightly outperforming RNAdvance Viral XP (Beckman, BM) in sensitivity of positive cases detection as well as reduced void samples (Supplemental Table 3).…”

Section: Homebrew Matches Commercial Extraction Of Rna For Saliva Sar...mentioning

confidence: 99%

“…Academic institutions became hubs where scientists changed their day-to-day research and/or teaching jobs and contributed to testing in multiple countries worldwide [4][5][6][7][8][9] . Our institution was no exception and we developed novel protocols based on reagents used in molecular research labs 2 , repurposed research spaces into diagnostic facilities 10 and with many others contributed with economic, accessible and sensitive methods to allow testing in resource-limited settings [11][12][13][14][15] . Universities have, proportionally, a large relatively young population (students), which had less risk of developing acute or severe COVID-19 as compared with older adults.…”

Section: Introductionmentioning

confidence: 99%

KCL TEST: an open-source inspired asymptomatic SARS-CoV-2 surveillance programme in an academic institution

Reis de Andrade,

Scourfield,

Peswani-Sajnani

et al. 2023

Preprint

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Testing was paramount in the management of the COVID-19 pandemic. Rapid deployment of new laboratories became widespread worldwide. Our university established KCL TEST: a SARS-CoV-2 asymptomatic testing programme that enabled sensitive and accessible PCR testing of SARS-CoV-2 RNA in saliva at 20 to 50% the price of commercial kits. We performed 158,277 PCRs in saliva for staff, students and their household contacts of Kings College London, free of charge and with an average turnaround time of 8 hours. Our pipeline is mainly made of open-source automation and non-commercial reagents and has been recently recommended for ISO15189 accreditation. Here we provide our blueprint and results to enable the rapid launch of diagnostic laboratories where and when needed. Our data span over 18 months and parallels that of the UK Office for National Statistics, with a lower positive rate and virtually no delta wave. Our observations strongly support regular asymptomatic community testing to decrease outbreaks and provide safe working spaces. KCL TEST demonstrates that universities can provide agile, resilient and accurate testing that reflects the infection rate and trend of the general population. We call for the integration of academic institutions in pandemic preparedness, with capabilities to rapidly deploy highly skilled staff, as well as develop, test and accommodate efficient low-cost pipelines.

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“…Isopropyl alcohol (or isopropanol, IPA) was added to enable DNA-silica interaction to detect human papillomavirus (HPV) from cervical swabs using a hand-sized fully integrated microfluidic device [112]. The increased binding efficiency was observed with about a 0.6 cycle improvement in the RT-qPCR test in the presence of 50% IPA and 1.25 M NaCl during the RNA-silica powder (glassmilk) binding process [123]. Binding buffers combining both Gu-based reagents and alcohol are also frequently reported.…”

Section: Reagents Anionic Supports Under Chaotropic Conditionsmentioning

confidence: 99%

“…Chaotropic agents are versatile as they can serve both cell lysis and NA binding, but the potential inhibitory effect of the chaotropes during amplification remains a concern. Moreover, the global shortage of GuSCN, which also has storage challenges as a toxic gas, hydrogen cyanide (HCN), can be formed when in contact with acid [128], places a great impetus on the development of alternative binding chemistries without the involvement of these chaotropes [123].…”

Section: Reagents Anionic Supports Under Chaotropic Conditionsmentioning

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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Homebrew: An economical and sensitive glassmilk-based nucleic-acid extraction method for SARS-CoV-2 diagnostics

Cited by 7 publications

References 23 publications

Moving towards on-site detection of Shiga toxin-producing Escherichia coli in ready-to-eat leafy greens

Moving towards on-site detection of Shiga toxin-producing Escherichia coli in ready-to-eat leafy greens

KCL TEST: an open-source inspired asymptomatic SARS-CoV-2 surveillance programme in an academic institution

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

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