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Inactivation of three emerging viruses – severe acute respiratory syndrome coronavirus, Crimean–Congo haemorrhagic fever virus and Nipah virus – in platelet concentrates by ultraviolet C light and in plasma by methylene blue plus visible light

  • DRK Blutspendedienst NSTOB, Springe, Germany
  • BRK Blutspendedienst, Nürnberg, Germany


Background: Emerging viruses like severe acute respiratory syndrome coronavirus (SARS-CoV), Crimean-Congo haemorrhagic fever virus (CCHFV) and Nipah virus (NiV) have been identified to pose a potential threat to transfusion safety. In this study, the ability of the THERAFLEX UV-Platelets and THERAFLEX MB-Plasma pathogen inactivation systems to inactivate these viruses in platelet concentrates and plasma, respectively, was investigated. Materials and methods: Blood products were spiked with SARS-CoV, CCHFV or NiV, and then treated with increasing doses of UVC light (THERAFLEX UV-Platelets) or with methylene blue (MB) plus increasing doses of visible light (MB/light; THERAFLEX MB-Plasma). Samples were taken before and after treatment with each illumination dose and tested for residual infectivity. Results: Treatment with half to three-fourths of the full UVC dose (0·2 J/cm2 ) reduced the infectivity of SARS-CoV (≥3·4 log), CCHFV (≥2·2 log) and NiV (≥4·3 log) to the limit of detection (LOD) in platelet concentrates, and treatment with MB and a fourth of the full light dose (120 J/cm2 ) decreased that of SARS-CoV (≥3·1 log), CCHFV (≥3·2 log) and NiV (≥2·7 log) to the LOD in plasma. Conclusion: Our study demonstrates that both THERAFLEX UV-Platelets (UVC) and THERAFLEX MB-Plasma (MB/light) effectively reduce the infectivity of SARS-CoV, CCHFV and NiV in platelet concentrates and plasma, respectively.
Vox Sanguinis (2020)
ORIGINAL PAPER ©2020 International Society of Blood Transfusion
DOI: 10.1111/vox.12888
Inactivation of three emerging viruses severe acute
respiratory syndrome coronavirus, CrimeanCongo
haemorrhagic fever virus and Nipah virus in platelet
concentrates by ultraviolet C light and in plasma by
methylene blue plus visible light
Markus Eickmann,
Ute Gravemann,
Wiebke Handke,
Frank Tolksdorf,
Stefan Reichenberg,
Thomas H. M
Axel Seltsam
Institute for Virology, Philipps University Marburg, Marburg, Germany
German Red Cross Blood Service NSTOB, Springe, Germany
Maco Pharma International GmbH, Langen, Germany
Received: 27 September 2019,
revised 18 December 2019,
accepted 18 December 2019
Background Emerging viruses like severe acute respiratory syndrome coronavirus
(SARS-CoV), CrimeanCongo haemorrhagic fever virus (CCHFV) and Nipah virus
(NiV) have been identified to pose a potential threat to transfusion safety. In this
study, the ability of the THERAFLEX UV-Platelets and THERAFLEX MB-Plasma
pathogen inactivation systems to inactivate these viruses in platelet concentrates
and plasma, respectively, was investigated.
Materials and methods Blood products were spiked with SARS-CoV, CCHFV or
NiV, and then treated with increasing doses of UVC light (THERAFLEX UV-Plate-
lets) or with methylene blue (MB) plus increasing doses of visible light (MB/light;
THERAFLEX MB-Plasma). Samples were taken before and after treatment with
each illumination dose and tested for residual infectivity.
Results Treatment with half to three-fourths of the full UVC dose (02 J/cm
reduced the infectivity of SARS-CoV (34 log), CCHFV (22 log) and NiV (43
log) to the limit of detection (LOD) in platelet concentrates, and treatment with
MB and a fourth of the full light dose (120 J/cm
) decreased that of SARS-CoV
(31 log), CCHFV (32 log) and NiV (27 log) to the LOD in plasma.
Conclusion Our study demonstrates that both THERAFLEX UV-Platelets (UVC)
and THERAFLEX MB-Plasma (MB/light) effectively reduce the infectivity of
SARS-CoV, CCHFV and NiV in platelet concentrates and plasma, respectively.
Key words: ultraviolet light, methylene blue, pathogen inactivation, plasma, plate-
let concentrates.
There is a large group of emerging viruses known to be
occasionally transmitted by blood or to have properties
suggesting their transmissibility by this route. These
pathogens include severe acute respiratory syndrome
coronavirus (SARS-CoV), CrimeanCongo haemorrhagic
fever virus (CCHFV) and Nipah virus (NiV), which have
been identified by the World Health Organization (WHO)
as major infectious threats with the potential to cause a
global pandemic [1–3].
There are different pathogen inactivation techniques
that have been developed to reduce or eliminate the
Correspondence: Axel Seltsam, German Red Cross Blood Service NSTOB,
Institute Springe, Eldagsener Strasse 38, 31832 Springe, Germany
Contributed equally to this work.
threat of infectivity from known and emerging transfu-
sion-transmissible agents [4]. THERAFLEX UV-Platelets
(Macopharma, Tourcoing, France) is a novel method for
pathogen inactivation treatment of platelet concentrates
(PCs) [5–7]. This purely physical system is based on short-
wave UVC light, which penetrates the fluid of PCs and
inactivates micro-organisms and leucocytes by damaging
nucleic acids. THERAFLEX MB-Plasma (Macopharma) is a
photodynamic pathogen inactivation procedure for treat-
ment of plasma [8,9]. Plasma units derived from single
blood donations are illuminated with visible light in the
presence of the phenothiazine dye methylene blue (MB).
When plasma is MB/light-treated, singlet oxygen is gen-
erated, which leads to the destruction of viral nucleic
acids. The MB/light-based method is in routine use in
Europe for about 17 years [10].
Both pathogen inactivation systems have been tested
in vitro to be effective against many different types of
viruses, including emerging viruses such as West Nile
virus and yellow fever virus [6,8,9,11–19]. In this study,
we investigated the capacity of THERAFLEX UV-Platelets
and THERAFLEX MB-Plasma systems to inactivate the
emerging viruses SARS-CoV, CCHFV and NiV in PCs and
plasma, respectively.
Materials and methods
Selection of donors
Selection of volunteer donors was based on local standard
practices. Only regular blood donors that fulfilled the
requirements for blood donation and had given their
informed consent approved by the local ethics committee
were included in the study.
Blood component preparation
Plasma-reduced PCs in platelet additive solution SSP
(Macopharma) were prepared from pools of five buffy
coats as previously described and were stored under agi-
tation at 22 2°C [14]. The target specifications of the
PCs were a platelet concentration of approximately
/mL and a plasma content of approximately 35% in
accordance with Macopharma’s specifications for UVC
treatment of PCs. Air was removed from all plasma units
and PCs.
Pathogen inactivation methods
Pathogen inactivation of PCs was performed using the
THERAFLEX UV-Platelets system (Macopharma) according
to the manufacturer’s instructions as described previously
[14]. All PCs were irradiated with UVC light to a total dose
of 02 joules per square centimetre (J/cm
) with constant
vigorous agitation to ensure uniform treatment [6].
Pathogen inactivation of plasma units was performed
using the THERAFLEX MB-Plasma system (Macopharma)
as described previously [14]. Plasma for pathogen inacti-
vation was processed by filtration for leucodepletion,
addition of MB and subsequent illumination with visible
light to a total dose of 120 J/cm
according to the
instructions of the manufacturer for this system [20]. The
MB removal step, an integral processing step in routine
use of the THERAFLEX MB-Plasma system, was omitted
for exclusive analysis of the virus inactivation effects of
Spiking experiments
Virus titres were determined by assessing for virus-induced
changes in morphology (cytopathic effects) of indicator
cells and calculated according to the SpearmanK
method and expressed as the log of the 50% tissue culture
infectious dose (log TCID50) [14,21,22]. Titration was per-
formed at the initial sample dilutions at which no cytotoxi-
city of indicator cells was observed. The effectiveness of
virus inactivation was calculated as the log reduction fac-
tor (RF) using the formula RF =log
, (R,
reduction factor; A
, spiked total virus load before treat-
ment; and A
, total virus load after treatment). The overall
reduction factor was expressed as the sum of RFs for all
steps. The limit of detection (LOD) of the assay was defined
as the lowest TCID
achievable at non-cytotoxic sample
SARS-CoV, strain Frankfurt 1 [23], was grown and
assessed in Vero E6 cells (ATCC CCL-22), CCHFV, strain
Afg09-2990 [24], was propagated and assessed in Huh7
cells (JCRB 0403), and NiV, strain Malaysia [25], was
grown and assayed in Vero 76 cells (ATCC CRL-1587).
For preparation of the virus stocks, viral supernatants
were collected on days 24 of cell culture, when a cell
confluence of approximately 80% was achieved, cen-
trifuged, aliquoted and frozen at -80°C until further use
in spiking experiments.
PCs (volume: 375 ml; n=2 per virus) and plasma
units (volume: 315 ml; n=2 per virus) were spiked 1:10
with supernatant of each virus and treated with UVC and
MB/light, respectively. After spiking, PCs and plasma
units were still within the specifications of the respective
pathogen inactivation method. The light doses were
applied incrementally until the full light doses of each
treatment were achieved. After each process step, samples
were collected and serially diluted for virus titration. In
order to test for intrinsic virus inactivation of the blood
product, reference samples were collected from each bag
before pathogen inactivation treatment, stored at room
©2020 International Society of Blood Transfusion
Vox Sanguinis (2020)
2M. Eickmann et al.
temperature and tested at the end of the experiments to
account for any intrinsic virus inactivation by the blood
The results of the infectivity assays demonstrated that
UVC irradiation and MB/light dose-dependently inacti-
vated SARS-CoV, CCHFV and NiV in plasma-reduced PCs
and plasma units, respectively. In PCs, at half of the full
UVC dose (01 J/cm
) SARS-CoV and CCHFV infectivity
levels were below the LOD, while at three-fourth of the
full UVC dose (015 J/cm
) also NIV infectivity levels
were below the LOD (Table 1). Thus, virus reduction fac-
tors 34 for SARS-CoV, 22 for CCHFV and 43 for
NiV were achieved with the UVC-based pathogen inacti-
vation system in PCs.
In plasma, already at one-fourth of the full light dose
(30 J/cm
) SARS-CoV, CCHFV and NiV were inactivated
to levels below the LOD (Table 2). These results corre-
spond to virus log reduction factors of 31, 32 and
27 that were achieved by MB/Light treatment for
SARS-CoV, CCHFV and NiV, respectively, in plasma.
For SARS-CoV, we observed a loss of infectivity of
about 1 log lower after spiking in some cases. This signif-
icant loss of infectivity was probably caused by non-
specific innate immune factors that neutralize viruses in
plasma [23,24]. However, virus titres did not further
decrease in controls during the course of the experiments.
In particular, there were no significant differences
between load and reference samples, indicating that the
observed virus inactivation was solely caused by the
treatment with UVC and MB/light.
A major argument for using pathogen inactivation tech-
nologies to treat blood components is that they support a
proactive approach providing more generalized protection
against new and emerging infectious agents which con-
tinuously challenge the safety of the blood supply. The
conventional reactive approach to wait until the threat
from an emerging transfusion-transmitted agent has been
identified before responding by modifying donor screen-
ing programmes takes time and, ultimately, the response
may not be quick enough to prevent the transfusion of
contaminated blood products. Because there are hundreds
of known emerging or re-emerging human pathogens
[26], the manufacturers of pathogen inactivation methods
are required to continuously test the inactivation capacity
of their systems for new infectious agents.
In this study, the inactivation efficacy of UVC and MB/
light was for the first time tested against CCHFV and NiV
Table 1 Inactivation of SARS-CoV, CCHFV and NiV in platelet concentrates by THERAFLEX UV-Platelets
Light dose (cumulative)
Bag 1 Bag 2 Bag 1 Bag 2 Bag 1 Bag 2
/ml log
RF log
/ml log
RF log
/ml log
RF log
/ml log
RF log
/ml log
RF log
/ml log
Virus stock 720169036802630175027403
Spiked bag 580360025203460262036502
005 J/cm
01 J/cm
015 J/cm
02 J/cm
Ref. sample 5902-015903015402-024902-036203-01630202
, 50% tissue culture infectious dose; RF, reduction factor; Ref. sample, pretreatment reference sample.
©2020 International Society of Blood Transfusion
Vox Sanguinis (2020)
Inactivation of SARS-CoV, CCHFV and Nipah virus 3
or other members of the Nairoviridae and Paramyxoviri-
dae families. SARS-CoV was also included in the study to
confirm the efficacy of the two pathogen inactivation
systems for coronaviruses, as has previously been demon-
strated for MERS-CoV [14]. The results of this study show
that both pathogen inactivation systems effectively inac-
tivated all three viruses spiked into the PC and plasma
samples, even at light dose increments below the full
doses recommended by the manufacturers. One limitation
of this study is that the number of replicates was small
due to safety constraints laboratory studies with these
zoonotic viruses must be performed in accordance with
the highest biosafety requirements. In addition, large-vol-
ume plating which would have allowed increasing the
sensitivity of the assay and consequently improving the
log reduction value could not be performed.
SARS-CoV is an enveloped, positive-sense single-
stranded RNA coronavirus. It emerged in 2002 in China
and spread to 29 additional countries and is thought to
be an animal virus that spread to humans from civets
most likely infected by bats [27]. Similar to Middle East
respiratory syndrome coronavirus (MERS-CoV), the main
route of human-to-human transmission of SARS-CoV is
nosocomial transmission. However, transmission between
family members has also been observed, suggesting that
SARS-CoV might continue to spread via transmission by
infected persons returning from affected areas. Transmis-
sion by blood transfusion has not been described yet.
Nevertheless, the high mortality of the disease and the
not yet fully understood transmission mechanisms of
SARS-CoV pose a potential threat to the safety of the
blood supply [27]. Interestingly, the detection of low-level
viremia in asymptomatic patients during an SARS-CoV
outbreak suggests a theoretical risk of transmission via
blood products. As a precautionary measure, the World
Health Organization introduced a recommendation for the
deferral of blood donations from donors potentially
exposed to SARS-CoV, and the Australian Red Cross
Blood Service amended its donor screening questionnaire
to include questions to identify persons with SARS-CoV-
related symptoms [28].
CrimeanCongo haemorrhagic fever virus is an envel-
oped, negative-sense single-stranded RNA virus of the
Nairoviridae family. CCHFV often results in a mild, non-
specific febrile illness but may occasionally cause severe
haemorrhagic disease. This disease was first identified in
the Crimean region of the former Soviet Union in 1944
and is a significant public health concern. CCHFV occurs
across a wide geographic region, including Europe, Asia
and Africa, and may expand into new regions [29]. The
virus is usually transmitted to humans through contact
with infected ticks and animal blood, but it is also trans-
missible from human to human via exposure to infected
Table 2 Inactivation of SARS-CoV, CCHFV and NiV in plasma by THERAFLEX MB-Plasma
Light dose (cumulative)
Bag 1 Bag 2 Bag 1 Bag 2 Bag 1 Bag 2
/ml log
RF log
/ml log
RF log
/ml log
RF log
/ml log
RF log
/ml log
RF log
/ml log
Virus stock 740271026503600275027502
Spiked bag 5403560352025102610.2 6402
30 J/cm
60 J/cm
90 J/cm
120 J/cm
Ref. sample 5403005703-015102015202-016202-01630201
, 50% tissue culture infectious dose; RF, reduction factor; Refsample, pretreatment reference sample.
©2020 International Society of Blood Transfusion
Vox Sanguinis (2020)
4M. Eickmann et al.
blood and other body fluids. Although no cases of CCHFV
transmission by blood transfusion have been reported to
date, incidences of hospital-acquired CCHFV infection
due to contaminated medical instruments have been doc-
umented [30]. These cases are strongly reminiscent of the
transmission routes of many other transfusion-relevant
NiV is an enveloped, single-stranded negative-sense
virus that belongs to the Paramyxoviridae family. It was
reported for the first time in the Malaysian population in
1998 and reappeared on different occasions in Asia. The
NiV disease spectrum ranges from asymptomatic infection
to acute respiratory illness and fatal encephalitis [31]. NiV
is a zoonotic virus transmitted to humans from animals
such as bats or pigs, but it can also be transmitted through
contaminated foods or directly person-to-person through
close contact with virus-containing body fluids and excre-
tions [32]. The available data, particularly the findings on
viral load in different body fluids, are too limited to pro-
vide a full understanding of its transmission routes [33].
Transfusion has not been implicated as a potential trans-
mission pathway to date. The incubation period of up to
14 days and the occurrence of latent infections with subse-
quent reactivation of NiV months and even years after
exposure suggest that infected persons may be overlooked
by donor screening programmes. However, the potential
transfusion risk may be limited by the fact that asymp-
tomatic and mild NiV infections are rare.
Future studies are needed to determine whether SARS-
CoV, CCHFV and/or NiV can be transmitted through
transfusion. If one or more of these viruses is transfusion
transmissible, its threshold concentration to elicit disease
must be examined to determine whether the capacity of
these pathogen inactivation technologies to inactivate the
respective virus in plasma and PCs is sufficient to prevent
transfusion transmission. Interpreting pathogen load in
relationship to infectivity and inactivation efficacy is
generally a very complex task [34]. When attempting to
do so, it is important to consider that because quantita-
tive polymerase chain reaction (qPCR), the most com-
monly used approach, measures viral load by detecting a
small fragment of the viral genome, the results may not
reflect infectivity and that qPCR usually overestimates the
titre of circulating infectious agents [34]. In contrast,
infectivity assays, which were used in this and previous
studies [5,12–14], determine the inactivation capacity of a
pathogen inactivation method based on intact, functional
viral units. Nevertheless, the log reduction factors
observed in this study and the safety margins calculated
from the inactivation levels achieved using only a frac-
tion of the standard light dose suggest that the THERA-
FLEX UV-Platelets and THERAFLEX MB-Plasma pathogen
inactivation technologies may effectively reduce the
potential risk of SARS-CoV, CCHFV and NiV and related
viruses for platelet or plasma transfusion.
We would like to thank Katharina Kowalski for excellent
technical assistance and the staff of the blood collection
and blood preparation departments for their support.
Conict of interest
FT and SR are employees of Macopharma, manufacturer
and distributor of the THERAFLEX pathogen inactivation
(PI) system. UG, WH, THM and AS received project grants
from the German Red Cross Blood Services and Maco-
pharma for the development of the UVC-based PI technol-
ogy for platelets. ME has no conflicts of interest to disclose.
Author contributions
M. Eickmann designed the study, interpreted the data and
co-wrote the manuscript. U. Gravemann designed the
study, performed the in vitro experiments and analysed
the data. W. Handke performed the in vitro experiments
and analysed the data. F. Tolksdorf interpreted the data
and edited the manuscript. S. Reichenberg interpreted the
data and edited the manuscript. T. H. M
uller interpreted
the data and edited the manuscript. A. Seltsam designed
the study, interpreted the data and wrote the manuscript.
All authors read and approved the final manuscript.
This work was supported by the German Red Cross Blood
Services (Deutsche Forschungsgemeinschaft der Blut-
spendedienste des Deutschen Roten Kreuzes) and Maco-
pharma S.A.S.
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... Examination of the inactivation of CCHF in biological samples revealed that the infectivity of plasma-reduced platelets was reduced by a factor of ‡2.2 log 10 to the limit of detection (LOD) when irradiated with ultraviolet C (UVC) at a wavelength of 254 nm to a dose of 0.1 J/cm 2 , whereas CCHV in plasma treated with methylene blue and light emitting diode-based light energy (30 J/cm 2 ) was reduced by a factor of ‡3.2 log 10 to below the LOD. 29,30 Irradiation with UV-A at 4000 lW/cm 2 at 20 min exposure 27 and (1200-3000 lW/cm 2 ) 28 has also proven effective. ...
... In PCs, at half of the full UVC dose (0.1 J/cm 2 ) SARS-CoV and CCHFV infectivity levels were below the LOD, while at three-fourth of the full UVC dose (015 J/cm 2 ) also NiV infectivity levels were below the LOD (Table 1). In plasma, already at one-fourth of the full light dose (30 J/cm 2 ) SARS-CoV, CCHFV and NiV were inactivated to levels below the LOD'' 29,30 AMT, 4¢-Aminomethyl-trioxsalen; CCHFV, Crimean Congo Haemorrhagic Fever virus; ID; LAIs, laboratory-acquired infections; LD; PFU, plaque forming units; TCID, tissue culture infectious doses; UV, ultraviolet; WHO, World Health Organization; NiV, Nipah virus; LOD, limit of detection. ...
... Coronaviruses are enveloped viruses and appear to be very sensitive to photodynamic inactivation [46]. The photodynamic activity of MB has been approved for several coronavirus species [12,30,47]. When BCoV coronavirus particles were incubated with 1 µM of MB for 10 min and irradiated with a 663 nm LED at a dose of 4 J/cm 2 , we clearly observed destructive effects on the virus envelope in electron micrographs [12]. ...
Full-text available
Methylene blue has multiple antiviral properties against Severe Acute Respiratory Syndrome-related Coronavirus 2 (SARS-CoV-2). The ability of methylene blue to inhibit different stages of the virus life cycle, both in light-independent and photodynamic processes, is used in clinical practice. At the same time, the molecular aspects of the interactions of methylene blue with molecular components of coronaviruses are not fully understood. Here, we use Brownian dynamics to identify methylene blue binding sites on the SARS-CoV-2 envelope. The local lipid and protein composition of the coronavirus envelope plays a crucial role in the binding of this cationic dye. Viral structures targeted by methylene blue include the S and E proteins and negatively charged lipids. We compare the obtained results with known experimental data on the antiviral effects of methylene blue to elucidate the molecular basis of its activity against coronaviruses.
... A significant challenge lies in the quantitative determination of the UV radiation dose required to inactivate pathogenic microorganisms (Gandhi et al., 2012;Feng et al., 2010). Although researchers have investigated the necessary dose of UV radiation to disinfect coronaviruses, it's noteworthy that the calculated and measured UV 50 dosages in these studies exhibit considerable variations (Walker and Ko, 2007;Buonanno et al., 2020;Tseng and Li, 2005;Terpstra et al., 2007;Pratelli, 2008;Deshmukh and Pomeroy, 1969;Eickmann et al., 2020;Kariwa et al., 2006;Kaur and Gupta, 2020). For instance, even within the 254 nm results, the log-reduction doses ranges widely: it is 0.6 mW·s·cm -2 for bovine coronavirus, while for SARS (CoV Urbani), it's as high as 11,754 mW·s·cm -2 (Heßling et al., 2020). ...
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The application of ultraviolet (UV)-based air disinfection holds promise, but also presents several 10 challenges. Among these, the quantitative determination of the required UV radiation dose for aerosols is particularly significant. This study explores the possibility of determining the UV dose experienced by aerosols without the use of virus-containing aerosols, circumventing associated laboratory safety issues. To achieve this, we developed a model system comprised of UV-sensitive dyes dissolved in di-ethyl-hexyl-sebacate (DEHS), which facilitates the generation of non-evaporating and UV-degradable aerosols. For the selection of UV-sensitive 15 dyes, 20 dyes were tested, and two of them were selected as most suitable according to several selection criteria. Dye-laden aerosol droplets were generated using a commercial aerosol generator and subsequently exposed to UVC radiation in a laboratory-built UV irradiation chamber. We designed a low-pressure impactor to collect the aerosols pre-and post-UV exposure. Dye degradation, as a result of UV light exposure, was then analyzed by assessing the concentration changes in the collected dye solutions using a UV-visible spectrophotometer. Our 20 findings revealed that a UV dose of 245 mW·s·cm-2 resulted in a 10% degradation, while a lower dose of 21.6 mW·s·cm-2 produced a 5% degradation. In conclusion, our study demonstrates the feasibility of using aerosol droplets containing UV-sensitive dyes to determine the UV radiation dose experienced by an aerosol.
... Ultraviolet light with the wave lengths of 100 nm to 280 nm (UV-C) has been widely used for the sterilisation of various materials, water and plant matter, with UV-C wave lengths of 250-260 nm being particularly effective to degrade and denaturate various viruses. [53][54][55][56][57][58] Numerous studies have shown that UV-C can also been successfully employed to degrade and denaturate SARS-Cov-2, [59][60][61][62][63][64][65][66] [67,68] and to degrade any other viruses or bio-hazardous agents that may have been harboured undetected by the users of the tests. Dosages greater than 13 mJ/cm 2 were found to be effective while a dosage of 16.9 mJ/cm 2 254 nm at was found to denaturate all SARS-Cov-2 viral material . ...
Technical Report
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The project ‘Material Culture of the COVID-19 pandemic’ aims to document various tangible manifestations of the pandemic. The aim of this document is to provide a formal documentation on a series of Rapid Antigen Tests that were acquired in Albury, NSW, or for use in Albury. All test sets described in this document have been handed to the Albury LibraryMuseum for accessioning into their local social history collection
... Previous studies of SARS-CoV susceptibility to 254 nm radiation, as well as other human and animal coronaviruses and, more recently, SARS-CoV-2 have indicated the effectiveness of 254 nm ultraviolet germicidal irradiation (UVGI) for inactivating various coronaviruses [10][11][12][13][14][15][16][17][18][19][20]. More recent studies have demonstrated the effectiveness of 222 nm radiation in inactivation of coronaviruses [12,13,21,22], but have also shown , relative to 254 nm and 222 nm radiation, the inefficiency of 270 nm and 282 nm radiation in inactivation of coronaviruses [12,13,23]. ...
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Ultraviolet germicidal irradiation (UVGI) is a highly effective means of inactivating many bacteria, viruses, and fungi. UVGI is an attractive viral mitigation strategy against coronaviruses, including the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the cause of the coronavirus disease-2019 (COVID-19) pandemic. This investigation measures the susceptibility of two human coronaviruses to inactivation by 254 nm UV-C radiation. Human coronavirus NL63 and SARS-CoV-2 were irradiated in a collimated, dual-beam, aqueous UV reactor. By measuring fluence and integrating it in real-time, this reactor accounts for the lamp output transients during UVGI exposures. The inactivation rate constants of a one-stage exponential decay model were determined to be 2.050 cm2/mJ and 2.098 cm2/mJ for the NL63 and SARS-CoV-2 viruses, respectively. The inactivation rate constant for SARS-CoV-2 is within 2% of that of NL63, indicating that in identical inactivation environments, very similar UV 254 nm deactivation susceptibilities for these two coronaviruses would be achieved. Given the inactivation rate constant obtained in this study, doses of 1.1 mJ/cm2, 2.2 mJ/cm2, and 3.3 mJ/cm2 would result in a 90%, 99%, and 99.9% inactivation of the SARS-CoV-2 virus, respectively. The inactivation rate constant obtained in this study is significantly higher than values reported from many 254 nm studies, which suggests greater UV susceptibility to the UV-C than what was believed. Overall, results from this study indicate that 254 nm UV-C is effective for inactivation of human coronaviruses, including SARS-CoV-2.
The characterization of modifications of microbial proteins is of primary importance to dissect pathogen lifecycle mechanisms and could be useful in identifying therapeutic targets. Attempts to solve this issue yielded only partial and non-exhaustive results. We developed a multidisciplinary approach by coupling in vitro infection assay, mass spectrometry (MS), protein 3D modelling, and surface plasma resonance (SPR). As a proof of concept, the effect of low UV-C (273nm) irradiation on SARS-CoV-2 spike (S) protein was investigated. Following UV-C exposure, MS analysis identified, among other modifications, the disruption of a disulphide bond within the conserved S2 subunit of S protein. Computational analyses revealed that this bond breakage associates with an allosteric effect resulting in the generation of a closed conformation with a reduced ability to bind the ACE2 receptor. The UV-C-induced reduced affinity of S protein for ACE2 was further confirmed by SPR analyses and in vitro infection assays. This comprehensive approach pinpoints the S2 domain of S protein as a potential therapeutic target to prevent SARS-CoV-2 infection. Notably, this workflow could be used to screen a wide variety of microbial protein domains, resulting in a precise molecular fingerprint and providing new insights to adequately address future epidemics.
Prior to the coronavirus disease-19 (COVID-19) pandemic, the germicidal effects of visible light (λ = 400 – 700 nm) were well known. This review provides an overview of new findings that suggest there are direct inactivating effects of visible light – particularly blue wavelengths (λ = 400 – 500 nm) – on exposed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virions, and inhibitory effects on viral replication in infected cells. These findings complement emerging evidence that there may be clinical benefits of orally administered blue light for limiting the severity of COVID-19. Possible mechanisms of action of blue light (e.g., regulation of reactive oxygen species) and important mediators (e.g., melatonin) are discussed.
Conference Paper
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Due to the recent pandemic, people are becoming conscious of the spread of germsdue to direct as well as indirect contact. The process of last mile delivery that is delivery directly to the house of customers from their nearby businesses and shops have suffered andreduced. This is due to no measures being taken to ensure cleanliness or sanitation during the process. The project described below is aimed at designing a robot capable of autonomous operation which will help with last mile delivery in India. This will help in providing consumers with a safe option for delivery. A compartment for storing the product,which also acts as the disinfection compartment is added to the product. The compartment will be equipped with disinfection mechanisms like UV Light disinfection, dehumidifying apparatus and disinfection spray to kill microbes present on the surface of the package.
Ultraviolet irradiation C (UVC) has emerged as an effective strategy for microbial control in indoor public spaces. UVC is commonly applied for air, surface, and water disinfection. Unlike common 254 nm UVC, far-UVC at 222 nm is considered non-harmful to human health, being safe for occupied spaces, and still effective for disinfection purposes. Therefore, and allied to the urgency to mitigate the current pandemic of SARS-CoV-2, an increase in UVC-based technology devices appeared in the market with levels of pathogens reduction higher than 99.9 %. This environmentally friendly technology has the potential to overcome many of the limitations of traditional chemical-based disinfection approaches. The novel UVC-based devices were thought to be used in public indoor spaces such as hospitals, schools, and public transport to minimize the risk of pathogens contamination and propagation, saving costs by reducing manual cleaning and equipment maintenance provided by manpower. However, a lack of information about UVC-based parameters and protocols for disinfection, and controversies regarding health and environmental risks still exist. In this review, fundamentals on UVC disinfection are presented. Furthermore, a deep analysis of UVC-based technologies available in the market for the disinfection of public spaces is addressed, as well as their advantages and limitations. This comprehensive analysis provides valuable inputs and strategies for the development of effective, reliable, and safe UVC disinfection systems.
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When the COVID-19 outbreak is ongoing, the classification of COVID-19 patients based on the severity assessment is necessary to optimize the allocation of existing resources and early management interventions to improve prognosis. Neutrophil-to-Lymphocyte Ratio (NLR) and Platelet-to-Lymphocyte Ratio (PLR) are two of the most common, simple, inexpensive, rapid, and widely available tests in all health facilities, which indirectly indicate the inflammatory status of COVID-19 patients. This study aimed to analyze the correlation between NLR and PLR with the severity of COVID-19 inpatients. This cross-sectional study was conducted retrospectively using medical record data of COVID-19 patients hospitalized at Al Islam Hospital, Bandung, from January to March 2021. COVID-19 patients involved in this study were classified into moderate, severe, and critical degrees. Statistical analysis was carried out using ANOVA or Kruskal-Wallis and Spearman with a significant value of p < 0.05. The median NLR and PLR results based on the severity were 3.49; 6.27; 8.4 (p<0.001) and 159.2; 202.6; 250.9 (p<0001), respectively. There was a correlation between NLR and PLR and the severity with r= 0.415 (p<0.001) and r=0.216 (p<0.001), respectively. The correlation between NLR and the severity was stronger than PLR. Therefore, it was concluded that there was a correlation between NLR and PLR with the severity of COVID-19 patients.
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Pathogen‐reduced (PR) platelets are routinely used in many countries. Some studies reported changes in platelet and red blood cell (RBC) transfusion requirements in patients who received PR platelets when compared to conventional (CONV) platelets. Over a 28‐month period we retrospectively analysed platelet utilisation, RBC transfusion trends, and transfusion reaction rates data from all transfused adult patients transfused at the Yale‐New Haven Hospital, New Haven, CT, USA. We determined the number of RBC and platelet components administered between 2 and 24, 48, 72 or 96 h. A total of 3767 patients received 21 907 platelet components (CONV = 8912; PR = 12 995); 1,087 patients received only CONV platelets (1578 components) and 1,466 patients received only PR platelets (2604 components). The number of subsequently transfused platelet components was slightly higher following PR platelet components (P < 0·05); however, fewer RBCs were transfused following PR platelet administration (P < 0·05). The mean time‐to‐next platelet component transfusion was slightly shorter following PR platelet transfusion (P = 0·002). The rate of non‐septic transfusion reactions did not differ (all P > 0·05). Septic transfusion reactions (N = 5) were seen only after CONV platelet transfusions (P = 0·011). These results provide evidence for comparable clinical efficacy of PR and CONV platelets. PR platelets eliminated septic transfusion reactions without increased risk of other types of transfusions with only slight increase in platelet utilisation.
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Background Nipah virus is a highly virulent zoonotic pathogen that can be transmitted between humans. Understanding the dynamics of person-to-person transmission is key to designing effective interventions. Methods We used data from all Nipah virus cases identified during outbreak investigations in Bangladesh from April 2001 through April 2014 to investigate case-patient characteristics associated with onward transmission and factors associated with the risk of infection among patient contacts. Results Of 248 Nipah virus cases identified, 82 were caused by person-to-person transmission, corresponding to a reproduction number (i.e., the average number of secondary cases per case patient) of 0.33 (95% confidence interval [CI], 0.19 to 0.59). The predicted reproduction number increased with the case patient’s age and was highest among patients 45 years of age or older who had difficulty breathing (1.1; 95% CI, 0.4 to 3.2). Case patients who did not have difficulty breathing infected 0.05 times as many contacts (95% CI, 0.01 to 0.3) as other case patients did. Serologic testing of 1863 asymptomatic contacts revealed no infections. Spouses of case patients were more often infected (8 of 56 [14%]) than other close family members (7 of 547 [1.3%]) or other contacts (18 of 1996 [0.9%]). The risk of infection increased with increased duration of exposure of the contacts (adjusted odds ratio for exposure of >48 hours vs. ≤1 hour, 13; 95% CI, 2.6 to 62) and with exposure to body fluids (adjusted odds ratio, 4.3; 95% CI, 1.6 to 11). Conclusions Increasing age and respiratory symptoms were indicators of infectivity of Nipah virus. Interventions to control person-to-person transmission should aim to reduce exposure to body fluids. (Funded by the National Institutes of Health and others.)
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Since emergence of the Nipah virus (NiV) in 1998 from Malaysia, the NiV virus has reappeared on different occasions causing severe infections in human population associated with high rate of mortality. NiV has been placed along with Hendra virus in genus Henipavirus of family Paramyxoviridae. Fruit bats (Genus Pteropus) are known to be natural host and reservoir of NiV. During the outbreaks from Malaysia and Singapore, the roles of pigs as intermediate host were confirmed. The infection transmitted from bats to pigs and subsequently from pigs to humans. Severe encephalitis was reported in NiV infection often associated with neurological disorders. First NiV outbreak in India occurred in Siliguri district of West Bengal in 2001, where direct transmission of the NiV virus from bats‐to‐human and human‐to‐human was reported in contrast to the role of pigs in the Malaysian NiV outbreak. Regular NiV outbreaks have been reported from Bangladesh since 2001 to 2015. The latest outbreak of NiV has been recorded in May, 2018 from Kerala, India which resulted in the death of 17 individuals. Due to lack of vaccines and effective antivirals, Nipah encephalitis poses a great threat to public health. Routine surveillance studies in the infected areas can be useful in detecting early signs of infection and help in containment of these outbreaks.
Background: To reduce the risk of transfusion transmission infection, nucleic acid targeted methods have been developed to inactivate pathogens in PCs. miRNAs have been shown to play an important role in platelet function, and changes in the abundance of specific miRNAs during storage have been observed, as have perturbation effects related to pathogen inactivation (PI) methods. The aim of this work was to investigate the effects of PI on selected miRNAs during storage. Study design and methods: Using a pool and split strategy, 3 identical buffy coat PC units were generated from a pool of 24 whole blood donors. Each unit received a different treatment: 1) Untreated platelet control in platelet additive solution (C-PAS); 2) Amotosalen-UVA-treated platelets in PAS (PI-PAS); and 3) untreated platelets in donor plasma (U-PL). PCs were stored for 7 days under standard blood banking conditions. Standard platelet quality control (QC) parameters and 25 selected miRNAs were analyzed. Results: During the 7-day storage period, differences were found in several QC parameters relating to PI treatment and storage in plasma, but overall the three treatments were comparable. Out of 25 miRNA tested changes in regulation of 5 miRNA in PI-PAS and 3 miRNA U-PL where detected compared to C-PAS. A statistically significant difference was observed in down regulations miR-96-5p on Days 2 and 4, 61.9% and 61.8%, respectively, in the PI-PAS treatment. Conclusion: Amotosalen-UVA treatment does not significantly alter the miRNA profile of platelet concentrates generated and stored using standard blood banking conditions.
BACKGROUND The THERAFLEX UV‐Platelets system (Maco Pharma) uses ultraviolet C (UVC) light for pathogen inactivation (PI) of platelet concentrates (PCs) without any additional photoactive compound. The aim of the study was to systematically investigate bacterial inactivation with this system under conditions of intended use. STUDY DESIGN AND METHODS The robustness of the system was evaluated by assessing its capacity to inactivate high concentrations of different bacterial species in accordance with World Health Organization guidelines. The optimal use of the PI system was explored in time‐to‐treatment experiments by testing its ability to sterilize PCs contaminated with low levels of bacteria on the day of manufacture (target concentration, 100 colony‐forming units/unit). The bacteria panel used for spiking experiments in this study included the World Health Organization International Repository Platelet Transfusion Relevant Reference Strains (n = 14), commercially available strains (n = 13), and in‐house clinical isolates (n = 2). RESULTS Mean log reduction factors after UVC treatment ranged from 3.1 to 7.5 and varied between different strains of the same species. All PCs (n = 12/species) spiked with up to 200 colony‐forming units/bag remained sterile until the end of storage when UVC treated 6 hours after spiking. UVC treatment 8 hours after spiking resulted in single breakthrough contaminations with the fast‐growing species Escherichia coli and Streptococcus pyogenes. CONCLUSION The UVC‐based THERAFLEX UV‐Platelets system efficiently inactivates transfusion‐relevant bacterial species in PCs. The comprehensive data from this study may provide a valuable basis for the optimal use of this UVC‐based PI system.
BACKGROUND Nonenveloped transfusion‐transmissible viruses such as hepatitis A virus (HAV) and hepatitis E virus (HEV) are resistant to many of the common virus inactivation procedures for blood products. This study investigated the pathogen inactivation (PI) efficacy of the THERAFLEX UV‐Platelets system against two nonenveloped viruses: HAV and feline calicivirus (FCV), in platelet concentrates (PCs). STUDY DESIGN AND METHODS PCs in additive solution were spiked with high titers of cell culture–derived HAV and FCV, and treated with ultraviolet C at various doses. Pre‐ and posttreatment samples were taken and the level of viral infectivity determined at each dose. For some samples, large‐volume plating was performed to improve the detection limit of the virus assay. RESULTS THERAFLEX UV‐Platelets reduced HAV titers in PCs to the limit of detection, resulting in a virus reduction factor of greater than 4.2 log steps, and reduced FCV infectivity in PCs by 3.0 ± 0.2 log steps. CONCLUSIONS THERAFLEX UV‐Platelets effectively inactivates HAV and FCV in platelet units.
BACKGROUND Ebola virus (EBOV) and Middle East respiratory syndrome coronavirus (MERS‐CoV) have been identified as potential threats to blood safety. This study investigated the efficacy of the THERAFLEX UV‐Platelets and THERAFLEX MB‐Plasma pathogen inactivation systems to inactivate EBOV and MERS‐CoV in platelet concentrates (PCs) and plasma, respectively. STUDY DESIGN AND METHODS PCs and plasma were spiked with high titers of cell culture–derived EBOV and MERS‐CoV, treated with various light doses of ultraviolet C (UVC; THERAFLEX UV‐Platelets) or methylene blue (MB) plus visible light (MB/light; THERAFLEX MB‐Plasma), and assessed for residual viral infectivity. RESULTS UVC reduced EBOV (≥4.5 log) and MERS‐CoV (≥3.7 log) infectivity in PCs to the limit of detection, and MB/light decreased EBOV (≥4.6 log) and MERS‐CoV (≥3.3 log) titers in plasma to nondetectable levels. CONCLUSIONS Both THERAFLEX UV‐Platelets (UVC) and THERAFLEX MB‐Plasma (MB/light) effectively reduce EBOV and MERS‐CoV infectivity in platelets and plasma, respectively.
Nipah virus, a Paramyxovirus related to Hendra first emerged in Malaysia in 1998. Clinical presentation ranges from asymptomatic infection to fatal encephalitis. Malaysia has had no more cases since 1999, but outbreaks continue to occur in Bangladesh and India. In the Malaysia-Singapore outbreak, transmission occurred primarily through contact with pigs while in Bangladesh-India, it is associated with ingestion of contaminated date palm sap and human-to-human transmission. Bats are the main reservoir for this virus which can cause disease in humans and animals. There are currently no effective therapeutics and supportive care and prevention are the mainstays of management.