Skip to Main Content
Page banner

Submitted
Being Assessed
In review
Responded
Thanks for your vote
This voting milestone has now ended, and the idea has progressed to a private milestone
Return to idea list
Thanks for approving the idea
This milestone has now ended, and the idea has progressed to a private milestone
Return to idea list

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

D2

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 understanding of lysis buffers and the need for high throughput COVID-19 testing led us to assess the possibility of extraction-free COVID-19 diagnosis. Our aim to produce a reagent that releases stable RNA, that is viable for RT-qPCR amplification and testing to be carried out without the need for costly and timely extraction methods. We also aim to overcome issues of extraction-free methods that can be hampered by the presence PCR inhibitors in Universal Transport Medium (UTM) and Viral Transport Medium (VTM)  creating problems with PCR diagnostic testing.

 MicroLYSIS-RNA

By modifying one of Microzone’s existing products we have developed a sample - lysis-direct to RT-qPCR lysis buffer. Our mixture of lysis reagents and PCR enhancement is providing very strong results. The buffer provides an extraction-free protocol for COVID-19 diagnosis.

Testing has demonstrated positive results using multiple RT-qPCR kits and real time thermal cyclers. To date we have analysis of a variety of samples including clinical sputum samples from a commercial collection container. We have also demonstrated post lysis RNA stability of greater than 48 hours with minimal loss of RNA when analysed using RT-qPCR.

Finally, the lysate has been used in RNA purification methods. Thus using our direct to PCR buffer, positive sample lysate can be later purified and taken to other downstream applications.

Ease of use – requires one tube (two tubes for dry swabs), vortex, short room temperature incubation and centrifuge step.

Implementation of this method would allow for a flexible approach to high-throughput testing that has previously been hampered by RNA extraction.

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

The method has been tested on numerous samples including clinical sputum. We are working to get more samples tested. Plus it has been tested using different RT-qPCR mastermixes and thermocyclers. It has so far proven to be very robust.

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

Very quick. We can supply MicroLYSIS-RNA for testing this week.

What is the likely production volume?​

We can increase production to in excess of 1 million reactions per week within a short time frame.

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

Not quite sure the risks and barriers that are being referred to. Producing at scale then risks and barriers are minimal. 

Who are you already partnering with on this?

No one presently although we have interested parties.

You will need to login to post a comment

Sonia Gandhi Apr 16, 2020

Status label added: C

Thanks for posting - do you have a plan to try microLYSIS on Covid samples, and is it compatible with inactivation/lysis buffer?

Reply 0

David Harris Apr 16, 2020

Hi Sonia,
Thank you for you comment and question.

MicroLYSIS-RNA has been tested on a COVID-19 Positive sputum sample. The results were very good. It came up strongly positive with a good CT. We are looking to test more COVID-19 samples this week.

Apologies, but I'm not clear what you mean by compatible with inactivation/lysis buffer. MicroLYSIS-RNA is a lysis buffer. If you would clarify the question, i'll be happy to answer by return.

David



Reply 0

Richard Hale Apr 16, 2020

David - is the MicroLYSIS buffer expected to inactivate viruses, or is it compatible with an additional heating step?

Richard

Reply 0

David Harris Apr 16, 2020

Hi Richard,

We expect it to inactivate the virus. However, we can not be completely certain of this as we can not test inactivation in our facilities. I can confirm that it is compatible with heating steps. We have heated sample to 90C and got good results. I would add that we do need to test this aspect on clinical samples. We are looking in to ways to achieve this at the moment.

Happy to have a further discussion on this topic.

David

Reply 0

Bev Matthews Apr 21, 2020

The idea has been progressed to the next milestone.

Reply 0

Bev Matthews Jun 4, 2020

Hello David, our champions have asked if you have additional validation data yet that they could review please? Look forward to hearing from you. Thank you bev

Reply 0

David Harris Jun 22, 2020

Hi Bev, apologies for the delay. Please find attached a pdf file that outlines sensitivity testing a quick validation trial run using MicroLYSIS RNA. We would welcome on going discussion about MicroLYSIS RNA and it use with Viral Transport Medium and Saliva. Also, happy to discuss any aspect of the information in the attached pdf. I look forward to hearing from you. David

Reply 0

Tom Jordan Jun 22, 2020

Thanks for this David, this is good timing. The champions were re-reviewing solutions earlier. On this one, they were interested in evidence on the inactivation and noted previous work focuses on sputum samples though that other samples might need testing. If it helps for you to share further details in relation to these two points please do. Though in the meantime I will note what you have shared to run by them.

Kind regards,
Tom

Reply 0

David Harris Jun 24, 2020

Hi Tom,
Thanks for the reply. On the two points you mentioned then I can add the following Saliva testing:

Further Saliva sample testing: We have run more in house testing using MicroLYSIS RNA, the Co-Diagnostics kit and SARS-CoV-2 controls re-hydrated with saliva. The results were very positive. See attached pdf.

Inactivation: We've approached a few groups. Still not managed to get any to move beyond an initial positive response. Any suggestions gratefully received. :)

Look forward to hearing from you.

Best wishes,
David

Reply 0

Tom Jordan Jul 1, 2020

Thanks for this David. We'll share this additional information with the review team. Tom

Reply 0

Tom Jordan Jul 23, 2020

Hi David,

Is there validation evidence using clinical samples in addition to control. Could we also double check on CE-IVD mark? With these details I can look into whether we can label this as a potential resource for labs.

Best wishes,
Tom

Reply 0

David Harris Jul 24, 2020

hi Tom,
Yes the product has been tested with clinical samples and it does work as outlined previously.
CE marking - working on this, however, as a general lab process we were informed that it doesn't need to be CE-IVD. Thoughts?

David

Reply 0

David Harris Sep 3, 2020

Not sure if anyone is monitoring these anymore. However, I did want to add that we two weeks ago we tested a positive sample. For the past two weeks the sample has stayed in the MicroLYSIS RNA buffer in a fridge. Today we tested again and the CT is teh same as it was when first tested. This demonstrates good stability of the RNA in our buffer.

Reply 0

Bev Matthews Jun 4, 2020

Status labels added: B: One to watch, More information requested

Status label removed: C: Further assessment

Reply 0

Tom Jordan Jul 9, 2020

Status label added: D2

Status labels removed: B: One to watch, Additional information

Reply 0

Share