Page banner

#TestingMethods2020

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

by Andrew Goldsborough | 9 months ago | in Catalogue of Resources | RNA extraction: New methods
D3: Catalogue of Resources

We have developed a virus inactivation and RNA stabilisation reagent called vivoPHIX™ 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.

Have you validated this method, if so, how and what were the results of the validation?​

Complete vivoPHIX™ validation of RNA stabilisation and inactivation of all tested viruses. 1 week validation of SARS-CoV-2 stabilisation, RNA purification and analysis by RT-qPCR (1 month stabilisation tests ongoing). Direct RT-qPCR analysis of vivoPHIX™ inactivated tissue culture cells carrying Influenza A H1N1 and one other RNA virus (no RNA purification step). We propose to extend these direct-to-cDNA results using vivoPHIX™ to stabilise and inactivate SARS-CoV-2 viruses making use of our understanding of compatibility between vivoPHIX™ and various cDNA products and platforms.

UPDATE: vivoPHIX has been successfully demonstrated to inactivate SARS-CoV-2 (preliminary results). Summary; all 9 tested viruses are inactivated by vivoPHIX.

How quickly could this be deployed and what are the dependencies?​

We have proven proof-of-principle for direct-to-cDNA for 2 RNA viruses, further vivoPHIX™ protocol tests needed for SARS-CoV-2 in swab and saliva samples.

What is the likely production volume?​

Protocol requires only small volumes of vivoPHIX™, no expected limitation of providing sufficient for requirements.

What are the risks and barriers to using this at scale?​

None known

Who are you already partnering with on this?

Various interested labs in Spain and the UK.

Files
INFA RNA
12 month vivoPHIX HCV stabilisation
SARS-CoV-2
Idea image
edited on Jul 1, 2020 by Andrew Goldsborough
Commenting is closed

Sonia Gandhi 9 months ago

Status label added: C

Bev Matthews 6 months ago

The idea has been progressed to the next milestone.

Tom Jordan 6 months ago

Status label added: D3: Catalogue of Resources

Status label removed: C: Further assessment

Tom Jordan 6 months ago

See below

Tom Jordan 6 months ago

The idea has been progressed to the next milestone.

Tom Jordan 6 months ago

Status label added: A: Not for current challenges

Status label removed: D3: Catalogue of Resources

Tom Jordan 6 months ago

I am closing this solution as there is an identical one here: https://testingmethods.crowdicity.com/post/3168120

Andrew Goldsborough 6 months ago

Hi Tom, unless I am mistaken this is not the same solution; using our reagent to stabilise SARS-CoV-2 and inactivate the virus followed by direct to cDNA (3390200) is different from using our reagent directly for RNA purification step followed by cDNA synthesis (3168120). Our aim was to respond to more than one challenge using our reagent, and the supporting data we uploaded to testing methods is different for the solutions. Thanks for your understanding.

Tom Jordan 6 months ago

Apologies for the misunderstanding Andrew, I will take a look at this now. Tom

Tom Jordan 6 months ago

Hi Andrew, I will bring this challenge back to the attention of the team so we consider it on the merit of it being a separate approach covering the RNA extraction challenge. My apologies for this. The team had the same query on applying this solution to COVID - I see you've answered this on the other solution you posted.

Tom

Andrew Goldsborough 6 months ago

Hi Tom, UPDATE: vivoPHIX has been successfully demonstrated to inactivate SARS-CoV-2 (preliminary results). Summary; all 9 tested viruses are inactivated by vivoPHIX. It seems that at least 2 of our submissions are now labelled as "A", was this intended?

Tom Jordan 6 months ago

Hi Andrew, thanks for this update, this is helpful. I have brought the current labels for VivoPHIX to the attention of the team so the current "A" label is being looked over and not final. I will update you on this as soon as I can. Is there any supporting information that might help to share with the reviewers alongside your update - do say if not or if you've attached this elsewhere? Thanks, Tom

Andrew Goldsborough 6 months ago

Hi Tom, after asking 9 different institutes to carryout the inactivation test including 5 in the UK, we received the SARS-CoV-2 inactivation data only yesterday from a major medical research institute in Australia. By its nature the inactivation result is preliminary and we will need to obtain authorisation from the institute to share the full results. I'll keep you posted but we now have 9/9 tested viruses inactivated with our reagent including Dengue, HepB, FIV, BVDV, Vaccinia, Zika, Influenza A, Rift Valley Forest Virus and (preliminary) SARS-CoV-2. Thanks Andy

Andrew Goldsborough 6 months ago

Tom, just as a reminder, we have SARS-CoV-2 sensitivity data (log2, log3 and log4) from a UK Lighthouse lab, as well as 33-day SARS-CoV-2 stability data. Is there anything else which the your team needs?

Tom Jordan 6 months ago

Thanks Andrew and apologies for the confusion. The reviewers looked at the additional information you provided and are keen to make this a resource on the platform for labs to consider for their use. Happy to discuss.

Kind regards,
Tom

Tom Jordan 6 months ago

Status label added: D3: Catalogue of Resources

Status label removed: A: Not for current challenges

Share