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WO2021223012A1 - Système à ultra-violet a (uva) et à ultra-violet c (uvc) et méthodes pour l'inactivation, la réduction et l'inhibition de la croissance du coronavirus - Google Patents

Système à ultra-violet a (uva) et à ultra-violet c (uvc) et méthodes pour l'inactivation, la réduction et l'inhibition de la croissance du coronavirus Download PDF

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Publication number
WO2021223012A1
WO2021223012A1 PCT/CA2021/050543 CA2021050543W WO2021223012A1 WO 2021223012 A1 WO2021223012 A1 WO 2021223012A1 CA 2021050543 W CA2021050543 W CA 2021050543W WO 2021223012 A1 WO2021223012 A1 WO 2021223012A1
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Prior art keywords
light source
uvc
uva
uvc light
uva light
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PCT/CA2021/050543
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English (en)
Inventor
Andrew Clark Baird AUBERT
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2449049 Ontario Inc
Helios Shield Ltd
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2449049 Ontario Inc
Helios Shield Ltd
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Priority claimed from PCT/CA2020/051059 external-priority patent/WO2021174331A1/fr
Application filed by 2449049 Ontario Inc, Helios Shield Ltd filed Critical 2449049 Ontario Inc
Priority to US17/299,378 priority Critical patent/US12458715B2/en
Priority to CA3177202A priority patent/CA3177202A1/fr
Priority to EP21799470.6A priority patent/EP4146289A4/fr
Publication of WO2021223012A1 publication Critical patent/WO2021223012A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/24—Apparatus using programmed or automatic operation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
    • A61L2/02—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor using physical phenomena
    • A61L2/08—Radiation
    • A61L2/10—Ultraviolet radiation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2202/00—Aspects relating to methods or apparatus for disinfecting or sterilising materials or objects
    • A61L2202/10—Apparatus features
    • A61L2202/11—Apparatus for generating biocidal substances, e.g. vaporisers, UV lamps

Definitions

  • UVA ULTRA-VIOLET A
  • UVC ULTRA-VIOLET C
  • This disclosure relates to a system and method of inactivating, reducing and inhibiting growth of coronavirus, in public areas such as areas frequented by humans in public transit vehicles and the like, by the use of UVA and UVC light sources at levels detrimental to coronavirus but safe for animals, including mammals and humans.
  • HCoV-229E human coronavirus
  • HCoV-OC43 Middle East respiratory syndrome coronavirus
  • SARS-CoV severe acute respiratory syndrome coronavirus
  • SARS-CoV-229E The first human coronavirus (HCoV) strain called B814 was isolated from the nasal discharge of a patient with a common cold in 1965. More than 30 additional strains were subsequently identified including HCoV-229E that was named so after a student specimen coded 229E. HCoV-229E was isolated by using the standard tissue culture method.
  • HCoVs including HCoV-299E strain can be responsible for 15%-30% of common cold cases in human adults. However severe respiratory tract infections may also occur in elderly people, infants or immunocompromised patient. Exposure to ultraviolet (UV) light can lead to antimicrobial activity. Far-UV light (for instance, from 207 to 222 nm) may be used as an efficient germicidal approach for killing microorganisms. UVC was found to provide the strongest antimicrobial activity among other types of UV radiation. For instance, it has been reported that far-UVC light (222 nm) inactivated airborne influenza virus. However, the exposure to UVC lamp might be associated with a health risk such as eye and skin damage.
  • UVC lamp might be associated with a health risk such as eye and skin damage.
  • UVC and UVB could be absorbed by RNA or DNA molecules and induce photo-chemical fusion of the adjacent pyrimidines into covalent-linked dimers such as thymine/cytosine dimers in DNA or uracil/cytosine dimers in RNA.
  • UV light may also damage RNA protein cross-linking, energy transfer between two proteins and result in site-specific damage to RNA.
  • UVA can provide oxidative damage to DNA, lead to production of reactive oxygen species and induce membrane damage.
  • external UVA (315-400 nm) and UVB (280-315 nm) are approved by FDA to use for the indication of eczema, psoriasis, skin lymphoma. UV light sources are known to be very effective in reducing coronavirus levels on surfaces. However, the typical radiated power and exposure time needed to reduce the levels of coronavirus may be deleterious to human eyes and epidermis and dermis layers.
  • an alternating UVA/UVC system for inactivating, reducing and inhibiting further growth, on a surface, of coronavirus, in one alternative, human coronavirus (HCoV-229E), wherein said system has no deleterious effects on an animal, including a human, in particular on a human eye or epidermis and dermis, wherein said system comprises: i) at least one UVA light source; ii) at least one UVC light source; and iii) at least one controller connected to each of said at least one UVA light source and said at least one UVC light source, for controlling at least one parameter of each of said UVA light source and UVC light source selected from light source, light intensity, radiated power level, wavelength, exposure time and combinations thereof; wherein said at least one UVC light source emits UVC light to a surface for a period of time reducing the level of said coronavirus on said surface to a level that is safe to animals including humans, and said at least one
  • said at least one UVC light source has an operating wavelength of from about 275 nanometers (nm) to about 295 nm. In one alternative, said at least one UVC light source has an operating wavelength of about 275 nm. [0006] According to one alternative, said at least one UVA light source has an operating wavelength of from about 385 nm to about 405 nm. In one alternative, said at least one UVA light source has an operating wavelength of about 405 nm.
  • said at least one UVC light source is a light emitting diode (LED).
  • said at least one UVA light source is a LED.
  • the at least one controller automatically cycles between emitting light from said at least one UVA light source and from said at least one UVC light source and said blanking time.
  • said at least one UVC light source has an emission at a power level and time duration to reduce coronavirus levels on a surface exposed to said at least one UVC light source.
  • the power level is selected to ensure the radiated emission from said at least one UVC light source is at a safe level for human eyes and epidermis and dermis.
  • the time duration is selected to ensure the radiated emission from said at least one UVC light source is at a safe exposure time for human eyes and epidermis and dermis.
  • said at least one UVA light source has an emission at a power level to inhibit growth of coronavirus on a surface exposed to said at least one UVC light source, while safe for human eyes and epidermis and dermis, regardless of the exposure time.
  • said at least one UVC light source has a power rating of from about 10 mW to about 100 W. In one alternative, said at least one UVC light source has a power rating of 236 mW.
  • said at least one UVA light source has a power rating of from about 10 mW to about 100 W. In one alternative, said at least one UVA light source has a power rating of 74 mW.
  • said system reduces the level of active coronavirus on a surface exposed to said system by 1 to about 100%. In one alternative, by 10 to about 20%.
  • a method of inactivating, reducing levels, on a surface, and inhibiting further growth of coronavirus, on said surface wherein said method has no deleterious effects on an animal, including a human, in particular on a human eye or epidermis and dermis, wherein said method comprises: i) Exposing said surface to at least one UVC light source for a period of time to reduce the level of coronavirus on said surface; ii) Terminating the exposure of the at least one UVC light source on said surface; iii) Exposing said UVC exposed surface to at least one UVA light source for a period of time to inhibit growth of said coronavirus on said surface; iv) Terminate the exposure of the at least one UVA light source on said surface; v) Providing a period of blanking time wherein said at least one UVA light source and said at least one UVC light source are off; vi) Optionally repeating steps i) to v) in order to maintain
  • said at least one UVC light source has an operating wavelength of from about 275 nanometers (nm) to about 295 nm. In one alternative, said at least one UVC light source has an operating wavelength of about 275 nm.
  • said at least one UVA light source has an operating wavelength of from about 385 nm to about 405 nm. In one alternative, said at least one UVA light source has an operating wavelength of about 405 nm.
  • said at least one UVC light source is a light emitting diode (LED).
  • said at least one UVA light source is a LED.
  • steps i) to v) are controlled by at least one controller automatically cycling between emitting light from said at least one UVA light source and from said at least one UVC light source and providing said blanking time.
  • said at least one UVC light source has an emission at a power level and time duration to reduce coronavirus on a surface exposed to said at least one UVC light source.
  • the power level is selected to ensure the radiated emission from said at least one UVC light source is at a safe level for human eyes and epidermis and dermis.
  • the time duration is selected to ensure the radiated emission from said at least one UVC light source is at a safe exposure time for human eyes and epidermis and dermis.
  • said at least one UVA light source has an emission at a power level to inhibit growth of coronavirus on a surface exposed to said at least one UVC light source, while safe for human eyes and epidermis and dermis, regardless of the exposure time.
  • said at least one UVC light source has a power rating of from about 10 mW to about 100 W. In one alternative, said at least one UVC light source has a power rating of 236 mW.
  • said at least one UVA light source has a power rating of from about 10 mW to about 100 W. In one alternative, said at least one UVA light source has a power rating of 74 mW.
  • said method reduces the level, and in another alternative inhibits growth, of active coronavirus on a surface by 1 to 100%. In one alternative, by at least one of the following ranges: 10 to 20%, 20 to 30%, 30 to 40%, 40 to 50%, 50 to 60%, 60 to 70%, 70 to 80%, 80 to 90% and 90 to 100%.
  • said method includes said at least one UVC light source is on for about 6 seconds, then off and followed immediately by at least one UVA light source on for about 6.5 hours, then both said at least one UVC light source and said at least one UVA light source off for about 1.5 hours for a blanking period before recommencing cycling of UVC and UVA light exposure, as required.
  • the UVA light source may remain on at levels safe to animals including humans to inhibit coronavirus growth and UVC is turned on at intervals to reduce coronavirus levels should coronavirus growth inhibition meet its limit, if any.
  • said system and method with blanking intervals are considered Risk exempt when tested to the IEC 62471 standard.
  • coronavirus may include FICoV-229E.
  • surface includes surfaces typically found in public places such as bathrooms and kitchens, including but not limited to countertops, hard counters, wood counters, concrete, plastic, rubber, leather, material and the like.
  • Figure 1 is a block diagram of the system, according to one alternative.
  • Figure 2 is a block diagram of the system, according to another alternative.
  • Figure 3a is a schematic of the setup for Example 1
  • Figure 3b is a photograph of the interior of the setup for Example 1.
  • Figure 4 is a schematic representation of the serial dilution carried out for Example 1.
  • Figure 5 depicts Non-infected (left) and infected (right) with HCoV-229E MRC- 5 cells of Example 1.
  • Figure 6 depicts the effect of UVA and UVC light on infectivity of HCoV-229E in 96-well plate of Example 1 following protocol 1.
  • Figure 7 Percentage of infected MRC-5 cells after 0, 1 and 3 cycles using protocol 1 in 96-well plate of Example 1 following protocol 1.
  • Figure 9 Percentage of infected MRC-5 cells after 0, 1 and 3 cycles in 96-well plate following protocol 2.
  • Figure 11 Percentage of infected MRC-5 cells after 0, 1 and 3 cycles in 96- well plate following protocol 2a.
  • Figure 12 Effect of protocol 1 , 2 and 2a on infectivity of HCoV-229E in 96-well plate.
  • Figure 13 depicts images showing MRC-5 cells at different experimental stages: non-infected control (a), infected cells with HCoV-229E before UV treatment (b), infected cells after 1 cycle of UV treatment (c) and infected cells after 3 cycles of UV treatment (d) using protocol 2a.
  • Figure 14 Effect of protocol 1 , 2 and 2a on infectivity of HCoV-229E in 24-well plate. Control is considered as “0 cycles”. Data shown represent mean of two independent experiments with error bars of standard deviation. Control was compared to treatments with P-values being ⁇ 0.5, >0.1 and ⁇ 0.01 for protocol 1 , 2 and 2a, respectively.
  • Figure 15 Effect of protocol 1 , 2 and2a on infectivity of HCoV-229E in 24-well plate. Control is considered as “0 cycles”.
  • FIG. 1 there is depicted a block diagram of a two continuous Pulse Width Modulation (PWM) example of one alternative for the system described herein.
  • a PWM generator 10 generates a continuous PWM which feeds into two circuits 20 and 30.
  • the first circuit is an optional logic buffer circuit 20 for controlling the pulsing of the UVC emitter 40.
  • the logic buffer circuit 20 ensures that the UVC emitter 40 is emitting when the PWM generator 10 is outputting a high logic level, and off when the PWM generator 10 is outputting a low logic level. See the output curve 22.
  • the second circuit is a logic inverter 30 which feeds into an OR circuit 40', along with the output of the second PWM generator 10', wherein the output of the OR circuit 40' controls the UVA emitter 50, ensuring that the UVA emitter is off when the PWM generator 10' is outputting a high logic level, and UVA emitter is on when the PWM generator 10' is outputting a low logic level. See the inverted output curve 32.
  • FIG. 2 there is depicted a block diagram of a three timer controlled system, according to one alternative.
  • UVC timer circuit 100 there is a UVC timer circuit 100, a UVA timer circuit 200 each controlling the UVC emitter 40 and UVA emitter 50 respectively, and a blanking timer circuit 300 for controlling the blanking period.
  • the UVC timer circuit 100 is set for 6 seconds on and the UVA timer circuit 200 is set for 6.5 hours and the blanking timer circuit 300 is set for 1.5 hours.
  • the UVC timer circuit 100 is enabled and outputs a logic high which is fed into a first logic buffer 110 and first logic inverter 120.
  • the first logic buffer 110 controls the UVC emitter 40 to be on with a high logic output and the UVC emitter 40 to be off with a low logic output, while the first logic inverter 120 is used to ensure the UVA timer circuit 200 is off.
  • the output changes state to turn off the UVC emitter 40 and turn on the UVA timer circuit 200 for 6.5 hours.
  • UVA timer circuit 200 outputs a logic high which is fed into a second logic buffer 210 and second logic inverter 220.
  • the second logic buffer 210 controls the UVA emitter 50 to be on, while the second logic inverter 220 is used to ensure the UVC timer circuit 100 is off.
  • the output changes state to turn off the UVA emitter 50 and turn on the blanking timer circuit 300 which ensures both UVC emitter 40 and UVC emitter 50 remain off for 1.5 hours, and the cycle repeats as required.
  • the 1.5 hour timer circuit 300 outputs a logic high and this output is fed into logic inverter 310 wherein the output of logic inverter 310 is combined with the output of logic inverter 220 and fed into logic AND circuit 60 producing a rising edge of the output of the logic AND circuit 60 which feeds in to the 6 second timer 100 to restart the 6 second timer 100 once the 1.5 hour timer circuit 300 completes the time.
  • the time value of each time may be determined by a variety of factors including, power level of UV light source, size of room, etc.
  • Example 1 UVA and UVC effect on coronavirus [00053]
  • the UV lamp was located in a sealed light box (See FIGS. 3a and 3b). Eyes were covered with a UV protective shield and a lab coat was worn at all times during the experiments. Experiments involving Human coronavirus (HCoV-229E) were carried out in a safety level 2 hood. Appropriate risk assessments and Control of Substances Hazardous to Health Regulations (COSHH) forms were completed prior to the experiment. Materials
  • MRC-5 (ATCC® CCL-171 TM) Human fibroblast cells were obtained from American Type Culture Collection and used to multiply HCoV-229E and for subsequent assays.
  • HCoV-229E (ATCC VR-740) was also purchased from American type culture collection.
  • Eagle's Minimum Essential Medium (EMEM) (ATCC® 30-
  • UVA (405 nm and 74 mW and 147 mW) and UVC (275 nm and 236 mW) light equipment was provided by Helios Shield LTD.
  • MRC-5 cells human lung fibroblast cells
  • Short-term frozen storage of the cells at -80°C was carried out by resuspending cell pellets produced by centrifugation for 5 minutes at 1200 rpm, in 1 ml of a solution of 900 pi of fetal calf serum (FCS) and 100 mI of dimethyl sulfoxide (DMSO).
  • FCS fetal calf serum
  • DMSO dimethyl sulfoxide
  • EMEM Eagle's minimum essential medium
  • Eagle's minimum essential medium contains Earle's Balanced Salt Solution, nonessential amino acids, 2 mM Glutamine, 1 mM sodium pyruvate, and 1500 mg/L sodium bicarbonate
  • the medium was supplemented with a mixture of penicillin- streptomycin antibiotic (1% of penicillin streptomycin antibiotic) and FCS (10% of FCS (fetal calf serum) to give a final concentration of 1 vol. % and 10 vol. %, respectively.
  • penicillin- streptomycin antibiotic 1% of penicillin streptomycin antibiotic
  • FCS fetal calf serum
  • the MRC-5 Cells were thawed (thawed from freezer to lab room temperature. Limited control. Thawing of frozen cells was performed as follows: a tube containing 1 ml of frozen cells was taken out of the freezer (-80°C) and left inside a tissue culture hood at room temperature (around 19°C). Once there was a small bit of ice left in the vial (usually after about a minute), transferred the cell suspension into a centrifuge tube and diluted to 1 :10 with EMEM medium, and centrifuged at 1200 rpm for 5 minutes. Pellets were then re-suspended in 1 ml of fresh EMEM medium, diluted to 1 :6 with the fresh EMEM medium, and incubated in 25 cm 2 tissue culture flasks at 37°C for up to 72 hours.
  • MRC-5 lung fibroblast cells were seeded at a concentration of 1 x 104 cell/ml into two 24-well plates 48 hours prior to the experiment.
  • the initial purchased stock of HCoV-229E in a volume of 100 pi was serially diluted to 10 -9 in EMEM media (serial dilution is shown in FIG. 4).
  • the old medium was replaced with each dilution of FICoV-229E in a fresh medium.
  • FIG. 3a the schematic representation of the experimental set up is shown.
  • the control 32 had two green buttons: the UVA 34 and UVC 36 switches.
  • the UV lamp 38 was placed 32 cm away from the 24-well plate 39 containing infected MRC-5 cells.
  • FIG. 3b is a photograph of the set up during the experiment with the UV lamp 38 and well plate 39.
  • UVC was activated for 6 seconds at the rotary position “F” (a light power level of 236 mW) and then deactivated.
  • UVA was then immediately activated for 6.5 hours at the rotary position “7” (light power level of 74 mW) and then deactivated.
  • the last 1.5 hours of the 8 hour interval was a blanking time, where both UVA and UVC light is off or deactivated.
  • Such UVC/UVA/blanking interval cycles were repeated up to 11 times. Viral inactivation was analysed after 1 , 3, 5, 7, 9, and 11 intervals as described below.
  • UVC was activated for 6 seconds at the rotary position “F” (a light power level of 236 mW) and then deactivated.
  • UVA was immediately pulsed (or activated) for 8 hours at the rotary position “F” (a light power level of 147 mW) and then deactivated. There was no blanking time and the UVC/UVA interval cycle was repeated. Such intervals were repeated up to 11 times. Viral inactivation was analysed after 1, 3, 5, 7, 9, and 11 intervals as described in below.
  • UVC was pulsed for 20 seconds at the rotary position “F” (a light power level of 236 mW) and then deactivated.
  • UVA was immediately pulsed for 8 hours at the rotary position “F” (a light power level of 147 mW) and then deactivated). There was no blanking time and the UVC/UVA interval cycle was repeated. Such intervals were repeated up to 11 times. Viral inactivation was analysed after 1, 3, 5, 7, 9, and 11 intervals as described in below.
  • the suspension containing infected cells, released virus and medium was transferred into a cryotube and underwent one rapid cycle of freeze and thaw, where the tube was placed for an hour at -80°C and subsequently thawed at room temperature for 30 minutes [B.-W. Kong, L. K. Foster and D. N. Foster, “A method for the rapid isolation of virus from cultured cells,” BoTechniques, vol. 44, pp. 1-5, 2018]. Then, the suspension was centrifuged at 2000 rpm for 10 minutes to remove cell debris and the culture supernatant. The supernatant was filter-sterilised using a 0.45 pm pore size filter and stored at -80°C until used for tissue culture infectious dose (TCID50) assay.
  • TCID50 tissue culture infectious dose
  • FICoV-229E untreated and treated for 1 , 3, 5, 7, 9, 11 cycles using either protocol 1 or protocol 2 or 2a was serially diluted in fresh EMEM medium.
  • 100 ⁇ l of virus suspension was placed into 900 pi of the fresh medium that was corresponded to 1 :10 dilution or 10 -1 as shown in FIG. 4 as per the protocol in S. E. Grimes, A Basic Laboratory Manual for the Small-Scale Production and Testing of I-2 Newcastle Disease Vaccine, RAP publications, 2002.
  • 100 pi of virus/medium suspension from 10 -1 was transferred to another tube containing 900 pi of fresh medium and classified as 10 -2 dilution or 1:100. This process was repeated to 10 '8 dilution factor.
  • MRC-5 cells at a concentration of 1 x 10 4 were seeded into either 96-well plate or 24-well plate. Once, the cells reached approximately 50% of confluence, they were infected with serially diluted treated coronavirus in 5 repeated wells for up to 4 days until cytopathic effect (CPE) was observed. Another plate was incubated with the serially diluted virus without any treatment in order to obtain control for tissue culture infectious dose (TCID50) and will be further called 0 cycles.
  • CPE cytopathic effect
  • TCID50 was calculated using the Reed and Muench method [L. J. Reed and H. Muench, “A simple method of estimating fifty per cent endpoints,” American Journal of Epidemiology, vol. 27, no. 3, pp. 493-497, 1938].
  • the formula for the calculations is the following (as per Reed and Muench): log10 50% endpoint dilution — tog10 of dilution showing a mortality next above 50% - (difference of logarithms X logarithm of dilution factor )
  • Cross-contamination risk will be calculated using the Exponential model, which represents a “dose-response” relationship between the dose applied to hosts (cells) and the probability of such a host to respond [T. Watanabe, T. A. Bartrand, M. H. Weir, T. Omura and C. N. Haas, “Development of a dose-response model for SARS coronavirus,” Risk Anal, vol. 30, pp. 1129-1138, 2010].
  • MRC-5 cells were infected with HCoV-229E and treated with UVA and UVC light as described.
  • Fig. 5 illustrates non-infected (left) and infected (right) MRC-5 cells, where CPE could be observed.
  • TCID50 assay was performed in order to investigate any infectivity of HCoV- 229E after each cycle of the treatment following protocol 1.
  • FIGS. 6 and 7 show the effect of protocol 1 on viral activity. As seen in FIG. 6, no CPE was observed after 1 cycle, whereas TCID50 was reduced from 5.1 log TCID50 to 2.5 log TCID50 after the first cycle. Control is considered as “0 cycles”. Data showed represent the mean of two independent experiments with error bars of standard deviation. Control (0 cycles) was compared to cycle one with P-value >0.05.
  • Protocol 2 was used for inactivation of HCoV-229E for up to 11 cycles. Data showed represent the mean of two independent experiments with error bars of standard deviation. Control (0 cycles) was significantly different to cycle 1 and cycle 3 (P-value ⁇ 0.01), respectively.
  • FIG. 8 shows the effect of each cycle on the ability of HCoV-229E to infect at least 50% of MRC-5 cells. TCID at 0 cycles was considered as the control and corresponded to 7.57 log TCID . As shown in FIG. 8, logTCIDso was reduced to 2.34 and 1.16 after the first and third cycle, respectively. No CPE was observed after 5, 6, 7 and 9 cycles.
  • TCID assay was also used in order to investigate any infectivity of HCoV- 229E after each cycle of the treatment following protocol 2a.
  • FIGS. 10 and 11 show the effect of UV lamp on CPE caused by HCoV-229E after 11 cycles. As illustrated in FIG. 10, TCID significantly reduced from 6.1 log TCID to 1.6 and 1.4 log TCID after 1 and 3 cycles, respectively. No CPE was observed after 3 cycles.
  • FIG. 10 there is shown the effect of UVA and UVC light on infectivity of HCoV-229E in 96-well plate.
  • the virus was treated with UV following the protocol 2.
  • Control is considered as “0 cycles”. Data showed represent the mean of two independent experiments with error bars of standard deviation. Control (0 cycles) was compared to cycle 1 and 3 with P-values being ⁇ 0.05 respectively.
  • FIG. 11 represents percentage of infected cells, which dropped from 100% to 60% and 40% at dilution 10 -1 after 1 and 3 cycles, respectively.
  • FIG. 12 shows the comparison of protocol 1 , 2 and 2a after 11 cycles, respectively, on infectivity of HCoV-229E in 96-well plate. Control is considered as “0 cycles”.
  • TCID50 reduced from the control to 1.6 log after 1 cycle using protocol B2.
  • TCID50 was 2.34 and 2.5 log after 1 cycle using protocols 1 and 2, respectively.
  • Flowever there was no CPE detected after 3 cycles using protocol 1 , whereas TCID50 was reduced to 1.6 and 1.4 log by protocols 2 and 2a, respectively.
  • protocol 1 might be the most successful setting in order to inactivate HCoV-229E after 3 cycles.
  • protocols 2 and 2a differed by the duration of UVC, the longer UVC treatment (20 seconds for protocol 2a) showed a slight change in TCID50 from 2.34 to 1.6 log.
  • FIGS. 13A-13D illustrates MRC-5 cells at different conditions: non-infected (FIG. 13A), infected, but not treated (FIG. 13B), infected and treated for 1 cycle (FIG. 13C) and infected and treated for 3 cycles (FIG. 13D). Images showing MRC-5 cells at different experimental stages: non-infected control (FIG. 13A), infected cells with HCoV-229E before UV treatment (FIG. 13B), infected cells after 1 cycle of UV treatment (FIG. 13C) and infected cells after 3 cycles of UV treatment (FIG. 13D) using protocol 2a. According to FIGS. 13A-13D, cells appeared rounded upon the infection, which was referred to CPE.
  • FIGS. 14 and 15 represent effect of different protocols on TCID50 of MRC-5 cells after 1 cycle in 24-well plates. According to FIGS. 14 and 15, the effect of protocols on CPE caused by HCoV-229E in 24-well plates was similar to 96-well plates.
  • FIG. 14 depicts the effect of protocol 1, 2 and 2a on infectivity of HCoV-229E in 24-well plate. Control is considered as “0 cycles”. Data shown represent mean of two independent experiments with error bars of standard deviation. Control was compared to treatments with P-values being ⁇ 0.5, >0.1 and ⁇ 0.01 for protocol 1 , 2 and 2a, respectively.
  • FIG. 15 depicts the effect of protocol 1, 2 and 2a on infectivity of HCoV-229E in 24-well plate. Control is considered as “0 cycles”. Cross-contamination
  • HCoV-229E could be infectious in a human lung cell culture such as MRC-5 for at least 5 days as well as on nonbiocidal surface materials: polytetrafluoroethylene, glass, polyvinyl chloride (PVC), silicone rubber, ceramic tiles and stainless steel [C. S. Heilingloh, U. W. Aufderhost, L. Schipper, U. Dittmer, O. Witzke, D. Yang, X. Zheng, K. Sutter, M. Trilling, M. Alt, E. Steinmann and A.
  • SARS-CoV-2 may be still infectious on surfaces such as on plastic surface for 3-4 days at a room temperature, SARS-CoV-1 can survive on the surface of polystyrene petri dish for at least 6 days at room temperature, but loss it's activity after 9 days [M. E. R. Darnell, K. Subbarao, S. M. Feinstone and D. R. Taylor, ''Inactivation of the coronavirus that induces severe acute respiratory syndrome, SARS-CoV,” J Virol Methods, vol. 121 , pp.
  • UVA was also used for viral inactivation. It was observed that UVA (540 ⁇ W/cm 2 at a distance of 3 cm) demonstrated weak inactivation of SARS-CoV-2 after 15 minutes, but UVC (1940 pW/cm 2 ) in a 400-fold decrease in infectious virus after 6 minutes [M. Bueckert, R. Gupta, A. Gupta, M. Garg and A.
  • UVC and UVA may be explained by mechanisms of light absorption. UVA light may be weakly absorbed by RNA and DNA and subsequently could be less effective in inducing pyrimidine dimers than either
  • UVC or UVB UVC or UVB.
  • UVA was found to cause additional genetic damage via production of reactive oxygen species that lead to oxidation of bases and strand breaks [M. Bueckert, R. Gupta, A. Gupta, M. Garg and A. Mazumder, “I nfectivity of SARS-CoV-2 and Other Coronaviruses on Dry Surfaces: Potential for Indirect Transmission,” Materials, vol. 13, pp. 1-16, 2020], [R. A, G. G. S. Leite, G. Y.

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  • Health & Medical Sciences (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
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  • General Health & Medical Sciences (AREA)
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Abstract

L'invention concerne un système à UVA/UVC destiné à réduire les taux actifs, sur une surface, et à inhiber la croissance supplémentaire du coronavirus sur ladite surface, ledit système n'ayant pas d'effets délétères sur un humain, en particulier sur l'œil humain ou l'épiderme et le derme humains, ledit système comprenant : iv) au moins une source de lumière UVA ; v) au moins une source de lumière UVC ; et au moins un dispositif de commande connecté à chaque source parmi ladite au moins une source de lumière UVA et ladite au moins une source de lumière UVC, pour commander au moins un paramètre de chaque source parmi la source de lumière UVA et la source de lumière UVC.
PCT/CA2021/050543 2020-03-03 2021-04-20 Système à ultra-violet a (uva) et à ultra-violet c (uvc) et méthodes pour l'inactivation, la réduction et l'inhibition de la croissance du coronavirus Ceased WO2021223012A1 (fr)

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US17/299,378 US12458715B2 (en) 2020-03-03 2021-04-20 Ultra-violet A (UVA) and ultra-violet C (UVC) system and methods for inactivation, reduction and inhibition of growth of coronavirus
CA3177202A CA3177202A1 (fr) 2020-05-04 2021-04-20 Systeme a ultra-violet a (uva) et a ultra-violet c (uvc) et methodes pour l'inactivation, la reduction et l'inhibition de la croissance du coronavirus
EP21799470.6A EP4146289A4 (fr) 2020-05-04 2021-04-20 Système à ultra-violet a (uva) et à ultra-violet c (uvc) et méthodes pour l'inactivation, la réduction et l'inhibition de la croissance du coronavirus

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US63/019,534 2020-05-04
CAPCT/CA2020/051059 2020-07-31
PCT/CA2020/051059 WO2021174331A1 (fr) 2020-03-03 2020-07-31 Système combiné d'ultra-violet a (uva) et d'ultra-violet c (uvc) pour la réduction et l'inhibition de la croissance d'agents pathogènes
US16/984,366 US12409245B2 (en) 2020-03-03 2020-08-04 Combination ultra-violet A (UVA) and ultra-violet C (UVC) system for reduction and inhibition of growth of pathogens
US16/984,366 2020-08-04

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