Case study : Virus disinfection on living human skin

Case study:
Disinfection testing on living human skin

Simultaneous visualization of virus disinfection efficiency and the impact on skin physiology

As an independant laboratory, we were contacted by the company Aquila Bioscience  to measure the efficiency of one of their disinfection product on living human skin. This ABD (Anti Bioagent Decontamination) Device works by physically capturing pathogens on both skin and surfaces. Pathogens bind to the solution and are stripped from skin and surfaces in a Velcro-like action. Our goal was to investigate and measure the efficacy of the product before commercial launch.

Note: our experiment on living human skin is conform to the requirements set out by the French Research Ethics Committee respecting the dignity, rights, safety and well-being of the people involved in the study.

Skin test with Abwipe

Challenging ABD wipes on SARS-CoV-2 contaminated skin

A prepartation of SARS-CoV-2 virus was deposit on the surface of living human skin explants. A small piece of a control wipe, PBS soaked wipe or ADB wipe was then apply without movement on the skin.

Determination of the skin decontamination efficiency of ABD device on SARS-CoV-2. A) SARS-CoV-2 titers in TCID50/ml for two biological replicates (R1 and R2). B) Log10 decrease of infectious SARS-CoV-2 for two biological replicates (R1 and R2).

After generation of a full report and a certificate of analysis for Aquila Bioscience customer, Abwipe was launched on the market.

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Case study : UVC disinfection

Case study:
UV-C disinfection box

Speeding up product development: fast calibration of UV-C power and exposure on autofluorescent viruses

As an independant laboratory, we were contacted by the company Clinit Technologies to measure the efficiency of one of their product, aiming at the disinfection of small objects. We used our autofluorescent ANCHOR™ Adenovirus type 5, a non-enveloped virus displaying high resistance to disinfection processes,  which allows simple and fast monitoring of viral reduction upon UV-C exposure. Results were generated in 24h. This viral model, which is quick to set up, is therefore the most relevant for defining the necessary decontamination times and power output to achieves the best infectious virus reduction.

UV-C disinfection of an ANCHOR™ hAdV5 (Green) as a function of exposure time.

Following several rounds of product optimization, validation on a collection of viruses interest

The commercial boxes developed by Clinit technologies aimed at the disinfection of at least 5 logs of SARS-CoV-2 and maximal effect on other class of viruses. To qualify the product, we used one of our SARS-CoV-2 strain in BSL3 laboratory and measure the efficiency of the final product directly before commercial release.

Clinit technologies product achieved high level of disinfection as measured directly on live infectious viruses:

           -SARS-COV-2: >99,9998% (>5log10)

           -Influenza virus H1N1: >99,998% (>4log10)

           -hAdenovirus5: >99,99% (>4log10)

We have also a unique capacity to visualize virus infection using advanced imaging techniques. For commercial and marketing applications, we also verified the efficiency of Clinit technologies product using microscopy.  On the slider below is shown SARS-CoV-2 infection before and after disinfection with Clinit product. (blue: cell nuclei, green SARS-CoV-2 viral RNA, red: SARS-Cov-2 NP).

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After generation of a full report and a certificate of analysis for Clinit customers, Clinit Technologies e-Box UV-C irradiation system was launched on the market.

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SARS-CoV-2

SARS-CoV-2

Characteristics

Severe Acute Respiratory Syndrome CorOnaVirus 2 is a member of the betacoronavirus family and triggers a disease called Covid19. The original strain (pre-VOC) emerged in the wuhan district in the late 2019 and rapidly spread over the world. Several variants of concern emerged, including the delta and omicron variant.

It is an enveloped virus (70-110nm) with a positive single stranded RNA genome of around 30kb.

Pathogenicity

In January 2022, more than 367 millions of people has been infected by SARS-CoV-2, triggering more than 5 million death. This virus is present worlwide.

  • Primary infection can be completely asymptomatic in healthy individuals
  • Most of people will develop low to mild-severe infection, including fever, dryed cough, fatigue, nausea and diarhea
  • Can trigger severe to fatal disease in patient, including hospitalization in ICU and artificial ventilation.
  • All antiviral in development to date failed to reach market entry, vaccine available that greatly reduce severity of symptomes
  • We use this virus for the discovery of new antiviral molecules, testing of neutralizing antibodies and measurement of disinfection procedures
  • We have also a strong library of protocol for the contamination of different surface of interest, from plastic to living human skin explants
NeoVirtech lab BSL3
All SARS-CoV-2 experiments are done in an accreditated BSL3 laboratory.
Virus screening NeoVirtech
SARS-CoV-2 destroying human pulmonary cells 48h post-infection. Nuclei in blue, SARS NP in red and SARS RNA in green

Applications

We use SARS-CoV-2 and other respiratory viruses for a broad category of testing. Contact us to know more.

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Orthopoxviruses ANCHOR™

Orthopoxviruses ANCHOR™

Characteristics

Orthopoxviruses are large enveloped virus, with a genome consisting of linear double-stranded DNA of 170 to 250 kilobases. Replication is strictly cytoplasmic

Pathogenicity

Pathogenic, can induce severe diseases in humans from variola virus (VARV virus eradicated in the 80s thanks to vaccination) to zoonotic monkeypox or cowpox virus..
  • Antivirals : Brincidofovir, Tecovirimat, Metisazone
HeLa cells infected with VacV-ANCHORTM, MOI 0.1, 48h PI, Cell Insight CX7 HCS System, ThermoFisher Scientific

Applications

Our Vaccinia virus (VacV) is combined with our ANCHORTM technology. This allows us to easily visualize viral infection and replication in high throughput imaging, making this virus an excellent tool. VacV-ANCHOR allows the discovery or repositioning of active antiviral molecules but also fundamental research on its biology. A Cowpox-ANCHOR virus is under development.

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Human Cytomegalovirus ANCHOR™

Human Cytomegalovirus ANCHOR™

Characteristics

Human Cytomegalovirus belongs to the Herpes virus family. It is a large enveloped virus (150-200nm), with a double-stranded DNA genome of around 240 kilobases.

Pathogenicity

Prevalence of 55-100% within human population depending on the geographic area and socioeconomic conditions:

  • Primary infection usually silent or benign in healthy individuals
  • Leading infectious cause of congenital abnormalities in western world (1-2,5% of birth defects)
  • Severe to fatal disease in immunocompromised individuals, organ transplant recipients or AIDS patients.
  • Main antivirals: Ganciclovir, Letermovir.  Ganciclovir and Letermovir resistant strains availables.
MRC5 cells infected with hCMV ANCHOR™ (green), MOI 0,5, 72h PI. Apotome ZEISS, 63X.
ANCHOR tagged virus (green) combined to anti-actin immunofluorescence (red), nuclei in blue

Applications

Our hCMV, hCMV Ganciclovir resistant and hCMV Letermovir resistant are combined with the ANCHOR™ technology. This allows us to easily visualize viral infection and replication in high throughput and in living cells, making this virus an excellent tool to investigate the behavior of hCMV infection in presence of a molecule of interest. This virus is used in routine for the discovery or repurposing of new antiviral molecules but also to understand the fundamental aspect of hCMV infection in living cells.

Reference:  Mariamé et al., Journal of Virology, 2018

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Human Adenovirus 5 ANCHOR™

Human Adenovirus 5 ANCHOR™

Characteristics

Human adenoviruses are medium-sized viruses (70 to 110 nm), with a genome consisting of double-stranded DNA (30,000 to 38,000 base pairs), with an icosahedral capsid and no enveloped.

Pathogenicity

Infection is usually silent during childhood. AdV infect epithelial cells.

  • 10% of respiratory infections are linked to an adenovirus
  • Adv is the causing agent of conjunctivitis, some serotype provoke severe keratoconjunctivitis (+++Japan)
  • Severe adenovirus infection in immunocompetent persons has been reported (CDC, 2000)
  • No antiviral available
sensible
hcs
no antiviral
Human Adenovirus 5 ANCHOR™ infecting cells in control conditions
Human Adenovirus 5 ANCHOR™ infecting cells on the presence of 35nM of an anti adenovirus compound

Applications

Our Adv5 is combined with our ANCHORTM technology. This allows us to easily visualize viral infection and replication in high throughput, making this virus an excellent tool.

Adv5 is a virus used in many applications, for the discovery or repositioning of active antiviral molecules but also an excellent model for measuring chemical, physical or radiative disinfection.

Remarkably, Adenoviruses are highly resistant to UVC because they can take advantage of the cell repair mechanisms of nucleic acid alterations1. This resistance, even though they are DNA viruses, reinforces the idea of using a labelled Adv5 as a model for rapid testing of many irradiation conditions.

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