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Completed- Fastgene versus pathogens aka RT PCR on chip: A valorization success story

Introduction to Fastgene technology

[Last Updated: April 15th, 2020]

pcr-on-chip covid-19 fastgene

Fastgene is a PCR-on-chip research project initiated at the beginning of Elveflow’s story. Taking advantage of microfluidics efficiency, this technology came out after almost a decade of research and development. 

It allows an ultra-rapid detection of pathogens (e.g., Ebola and the SARS-CoV-2 virus of the Covid-19 crisis).

pcr-on-chip biology

In 2010-2011, Elvesys’ founders worked on a disruptive project. What if temperature could be controlled at the second scale to allow unprecedented biological applications?

 

From this simple question, two main applications arose. On the one hand, a team led by Jeremy Cramer focused on controlling temperature to precisely control cell mechanisms (cell division, chromosome segregation, protein stability…). Cherry Biotech was founded in 2014.

Fast PCR-on-chip for pathogens detection

On the other hand, temperature ramps are the limiting factor in the gold standard PCR technique. Did you say PCR? Polymerase Chain Reaction is a gold standard used in laboratories to amplify DNA. With many genetic applications (cloning, detection, mutation…, etc.), PCR is a sequence of steps where an original DNA sequence is processed to allow for rapid duplication and amplification until a sufficient quantity of nucleic acid material is obtained.

 

If we simplify, a typical PCR cycle follows three steps:

  1. Denaturation of the original double-stranded DNA sequence. Typically, it is around 95°C for 20-30 seconds.
  2. Annealing of specific primer DNA short sequences to the single-stranded DNA matrix. From 50 to 65°C, depending on the primers, for approx. 30 seconds.
  3. Extension of the newly synthesized complementary DNA strand. Temperature of 72°C for some minutes, depending on the length of the DNA strand to be copied.

 

 

Steps 1 to 3 are repeated 25-30 times, allowing for an exponential amplification of the original DNA sequence. As each cycle doubles the DNA quantity, one may end up with 2^30 copies after 30 cycles!

Consequently, the limiting factor to speed up the entire process is the ability to get rapid thermal cycles. From there, started the “Fastgene” research pro project. This original temperature question is about the efficiency of low volume-to-surface ratios. 

A liquid whose temperature is tightly controlled can be actuated into a specific chamber, allowing for a precise and very efficient control of temperature. Controlling and changing the temperature at the micro-scale is easy and ultra-rapid, as shown in an original Fastgen quantitative PCR characterization characterization.

fastgene-microfluidic-pcr-on-chip-qPCR-qRTPCR

Mael Leberre founded BforCure in 2018 after a long period of research and development. The original technology has been improved and is now a robust system to detect short nucleic acid sequences in an unprecedented short time.

As a strong proof-of-concept, the Fastgene platform detected Ebola and Anthrax variants in less than 10 minutes! (see LOC publication).

Today, we are very proud of this story as BforCure was selected by the French Ministry of Armed Forces with a 1.8 million Euro grant to develop a rapid point-of-care method to detect COVID-19 in less than 30 minutes!

The project is called NOMORECOV and will strongly contribute to boosting BforCure’s objective: “For future outbreaks, including the possible seasonal reoccurrence of SARS-CoV-2, people will be able to be tested and have results within minutes at multiple sites, such as airports, hospitals, outside of office buildings, etc.”

 

We have a single word to say to the BforCure team: BRAVO!

PCR-on-chip for bacteria and viruses diagnosis in only 7 minutes

Aim of the project

pcr-on-chip to diagnose Ebola virus

Detect Ebola in 8 minutes? Anthrax in 7 minutes? It is possible with PCR-on-chip.

The Fastgene project aims to exploit the unique features of microfluidics to provide the world’s fastest qPCR system for medical diagnosis. Indeed, Fastgene can detect the presence of pathogens such as Bacillus anthracis (Anthrax agent) and the Ebola virus in less than 8 minutes (which means seven times faster than conventional off-the-shelf apparatus).

More importantly, this outstanding reduction in the assay duration does not affect qPCR amplification volume, efficiency, or sensitivity, which are on par with conventional apparatus.

Why is it so important to be fast and specific?

The 2014 Ebola outbreaks demonstrated that currently available diagnostic tools are not up to tackle effectively widespread epidemics.

Indeed, current devices are:

  • Either fast but lacking sensitivity, meaning they don’t allow early diagnosis.
  • Or sensitive but time-consuming, meaning they need several hours to produce a result that the doctor can process.
pcr-on-chip to diagnose anthrax virus

History of the Fastgene PCR-on-chip

Elveflow is committed to this goal: develop the fastest specific diagnostic tool.

fastgene-microfluidic-lab-on-chip-qPCR-qRTPCR_PCR DNA amplification






The Fastgene system allows ultra-fast assays typical of research-oriented setups while retaining the same performance of off-the-bench approaches regarding sensitivity and specificity. Such performances are made possible thanks to the most sensitive and reliable diagnostic method: the real-time Polymerase Chain reaction (qPCR).

From a PhD thesis to the most innovative system of 2014

Elvesys is a French startup in microfluidics founded in 2011. Since then, Elvesys has created a new startup every year. In 2012, it obtained funding from the DGA for Fastgene, managed in partnership with the ENS. In 2014, Elvesys was awarded the “Innovation 2030 Worldwide Challenge” thanks to Fastgene.

Fastgene PCR-on-chip system and its performances

PCR on chip: Fastgene’s powerful weapons against diseases.

Fastgene protoype qpcr

The Fastgene diagnostic system is based on the quantitative polymerase chain reaction (qPCR), a molecular test that detects bacterial and virus DNA. 


Fastgene technology uses a patented microfluidic thermalization technique with microfluidic chips and Elveflow high-performance pressure controllers.

Outstanding thermalization performances enables ultra-fast detection.

Fasgene thermalisation performances 30 pcr cycles in 2 min

Microfluidics thermalization is the basis of Fastgene, enabling it to be the fastest microfluidic-based diagnostic tool to our knowledge. 

Indeed, the Fastgene system has been tested on Anthrax and Ebola-simulating agents in 7 minutes, making it 7-14 times faster than current systems. 

Furthermore, the Fastgene detection limit is on par with off-the-shelf methods.

Temperature profile of Fastgene microfluidic chip
fastgene microfluidic chip qrtpcr sensitivity comparison
Molecular detection efficiency of the Fastgene system: despite being 6 times faster, the Fastgene system presents the same amplification efficiency as commercial state-of-the-art system. The Fastgene system presents also the same sensitivity (1,2).
fastgene microfluidic chip qrtpcr speed comparison
Molecular detection of Fastgene compared to a commercial state-of-the-art system: the Fastgene system enables a detection 6 times faster.

The people behind the study

Team-Elveflow

Mael

DR. MAËL LE BERRE

ELVESYS CTO

Phd in Microfluidics

Manager of the Project

Walter

WALTER MINNELLA

MARIE CURIE PHD FELLOW

Msc in Applied Physics

Responsible of thermics and fluidics

Mathilde

MATHILDE DUPARD

R&D ELECTRONICHS ENGINEER

Msc in Electrotechnics

Responsible of Electronics

DSC04063 (2)

DR. PIERRE-EMMANUEL DOUARRE

R&D MOLECULAR BIOLOGY ENGINEER

PhD in Microbiology

Responsible of molecular biology

Duc

DR. AHN DUC VU

R&D OPTICS ENGINEER

PhD in Physics

Responsible of optics

Morgan

MORGAN CAZAUD

INTERN

Msc in Electrotechnics

Mechanical conception

References

1. Houssin, T., Cramer, J., Grojsman, R., Bellahsene, L., Colas, G., Moulet, H., Minnella, W, Pannetier, C., Leberre, M., Plecis Chen, Y. (2016). Ultrafast, sensitive and large-volume on-chip real-time PCR for the molecular diagnosis of bacterial and viral infections. Lab Chip. doi:10.1039/C5LC01459J

2. D. Lee, P.-J. Chen and G.-B. Lee, Biosens. Bioelectron., 2010, 25, 1820–1824.

3. S. Kirchner, K. M. Krämer, M. Schulze, D. Pauly, D. Jacob, F. Gessler, A. Nitsche, B. G. Dorner and M. B. Dorner, Appl. Environ. Microbiol., 2010, 76, 4387–4395.

Elveflow has been awarded several times and covered in many national media outlets for its Fastgene technology.

Fastgene project presented for the TV show “Des inventions et des hommes” (“Inventions and Men”) on M6 channel.

fastgene-microfluidic-lab-on-chip-qPCR-qRTPCR_Fastgene-awarded-at-the-Innovation-2030-Worldwide-Chal
Fastgene awarded at the Concours Mondial d’Innovation 2030 (Innovation 2030 Worldwide Challenge) with the French Prime Minister
fastgene-microfluidic-lab-on-chip-qPCR-qRTPCR_Fastgene-awarded-and-presented-for-Grand-Prix-de-lInno
Fastgene awarded and presented for Grand Prix de l’Innovation Paris (Paris Innovation Grand Prize): “Make a difference: if you found out in less than 10 minutes the origin of your fever, it is also thanks to us”.

Corresponding press article in l’Entreprise here

Ultra fast temperature control PCR-on-chip for biology

Other press articles about the FASTGENE project

Press article about Fastgen in 20 minutes: “It is one of Hollywood film’s favorite subjects. If movies like Outbreak, The Flu, or Contagion show with or less realism the consequences of a pandemic on civil society, in real life, ELVESYS developed a technology enabling the detection of any virus or bacteria three times faster than any regular methods by using microfluidics.”


Press article in la Tribune about Fastgene here with an interview of ELVESYS President Guilhem Velve-Casquillas. This latter explains: “[The Fastgene project] is based on my thesis and is very important for me. It will detect any virus, bacteria, or tumors in less than 10 minutes, at lower costs, without going to the hospital.”


Press article in l’Usine Nouvelle about Fastgene PCR on chip here: “[ELVESYS] also develops with the Defense ministry a technology enabling a lab-on-chip to detect a virus or a bacteria in 10 minutes.”


Press article in Silex ID about Fastgene, dec. 2014, here: “[The Fastgene project] consists in a miniaturized chip able to “scan” the human organism in less than 10 minutes, from a single drop of blood or saliva. This microfluidic technology, adaptable to any lab-on-a-chip and enabling rapid detection of bacteria or even circulating tumor cells, is a potent prevention tool, notably in the case of bacteriological attack. The French army is, moreover, an ELVESYS’ purchaser”.