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RNA extraction: New Methods

23 Ideas
20 Votes
176 Comments
86 Subscribers

RNA extraction capacities are currently challenged even with automated platforms.

New methods of extracting viral RNA or enabling viral detection without an extraction step would help remove this bottleneck, as long as they are “ready to go” and can be integrated into existing or optimised PCR testing chains.

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B: One to watch

N-acetyl-L-cysteine to disrupt mucus and increase RNA extraction and sensitivity

A paper from 2004 showed addition of N-acetyl-L-cysteine to the sample could disrupt the mucus and increase the yield of RNA by ~2.75 times.

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Additional information

Viral RNA Lysis Buffer

Viral RNA Lysis Buffer is designed for: • Nasopharyngeal swab samples • Lysis of DTT pre-treated sputum samples which had been spun down to pellets before adding the RNA Lysis buffer • Stabilization of nucleic acids with lysates Compatible with commercially available RNA extraction kits comprising magnetic beads or spin columns, prior to RT-qPCR.

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E: A connection is helpful

Rapid Covid-19 IgM / IgG Point of Care Test

The IgG/IgM Rapid Test is based on rapid immunochromatographic test. It is used for the detection of 2019 Novel Coronavirus IgG and IgM antibody in human whole blood/serum/plasma, offering a diagnostic reference for COVID 19. IgG line uses a cocktail of antibodies against Protein N and Protein S.

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E: A connection is helpful

WHO APROVED COVID-19 IgG/IgM Rapid Test (colloidal gold-based) (CE-IVD) 10 Minutes result.

The idea is to with less Human Power and the already used knowledge of medical Personnel to implement this Testers which allow to have very accurate results within 10 minutes. We can provide them To any Airport to any border to any control station train the Medical Staff. I would recommend also Heat scanners to make the first broadband check. 

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D2

Increase Covid Testing by 3, 9 or 15 fold

Mass Testing for Covid 19 – Welch Protocol This depends on the actual level of Covid 19 in the community. Particularly useful where you expect the incidence of positives within a group to be low, such as all inhabitants and staff in a care home, NHS staff or mass testing of the population in general Results to date show that roughly 1 in 4 of those tested are positive, so far these are mainly patients with symptoms. At 1 in 4 you might as well test individual samples – see below...

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B: One to watch
Additional information

MicroLYSIS-RNA : From sample to lysis to RT-qPCR

Microzone, a division of Clent Life Science, has been manufacturing molecular biology products for over 20 years in the UK. Microzone has historical experience of developing direct to PCR lysis buffers. For example, Microzone was one of, if not the first, company to place a commercial reagent to go from sample to lysis to PCR in to the market. This product is called MicroLYSIS and still in use today. Subsequent development saw the release of MicroLYSIS Plus and DNAmite for plants. Our...

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C: Further assessment

Direct-to-PCR COVID-19 Testing - Enhancing sensitivity

The Novacyt Group specialises in molecular testing technologies. Primerdesign, a part of the Novacyt Group, has a track record of developing in vitro diagnostics and led the world in the assay development response to COVID-19.   Continuing these efforts, our research and development team have screened a variety of extraction-free viral testing protocols and are currently in the process of validating the lead candidate approach.   Based on our preliminary analytical data, we are...

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A: Not for current challenges
Additional information

RT-PCR detection directly from sample

Based on our patent WO2016106113 we believe that a direct from sample RT-PCR reaction can be used to detect Covid-19 without the need of an extraction step. This technology could be used on any open qPCR/RT-PCR equipment already existing on most of the labs. This will enable faster detection & eliminate the RNA extraction bottleneck. 

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A: Not for current challenges

vivoPHIX™ RNA stabilisation, virus inactivation and direct sample to cDNA protocol

We have developed a virus inactivation and RNA stabilisation reagent called  vivo PHIX™ that has been used for SARS-CoV-2 sample collection. We intend to use our reagent and know-how to replace the RNA purification step of the current SARS-CoV-2 test protocol with a ‘Direct-to-cDNA’ protocol, thereby reducing costs, avoiding reagents in short supply, and improving testing throughput.

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B: One to watch

Super high throughput testing using Next Generation Sequencing

This is not a method for RNA isolation but rather the replacement of testing by qPCR by a far more high throughput method – millions of tests in one experiment . When RNA isolation problem stops being a bottle neck in testing, the method proposed below will remove the need for qPCR, which, in turn, will become the new bottle neck. The scheme of the process is shown in attached Figure. - The reverse transcription primer contains sequence standard for Illumina flow cell (blue), followed...

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