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1.
Viruses ; 13(2)2021 02 02.
Article in English | MEDLINE | ID: covidwho-1067781

ABSTRACT

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spread globally. Although measures to control SARS-CoV-2, namely, vaccination, medication, and chemical disinfectants are being investigated, there is an increase in the demand for auxiliary antiviral approaches using natural compounds. Here we have focused on hydroxytyrosol (HT)-rich aqueous olive pulp extract (HIDROX®) and evaluated its SARS-CoV-2-inactivating activity in vitro. We showed that the HIDROX solution exhibits time- and concentration-dependent SARS-CoV-2-inactivating activities, and that HIDROX has more potent virucidal activity than pure HT. The evaluation of the mechanism of action suggested that both HIDROX and HT induced structural changes in SARS-CoV-2, which changed the molecular weight of the spike proteins. Even though the spike protein is highly glycosylated, this change was induced regardless of the glycosylation status. In addition, HIDROX or HT treatment disrupted the viral genome. Moreover, the HIDROX-containing cream applied on film showed time- and concentration-dependent SARS-CoV-2-inactivating activities. Thus, the HIDROX-containing cream can be applied topically as an antiviral hand cream. Our findings suggest that HIDROX contributes to improving SARS-CoV-2 control measures.


Subject(s)
Antiviral Agents/pharmacology , Olea , Phenylethyl Alcohol/analogs & derivatives , Plant Extracts/pharmacology , /drug effects , Administration, Topical , Animals , Antiviral Agents/chemistry , Carbohydrates/chemistry , Chlorocebus aethiops , Genome, Viral/drug effects , Glycosylation , Microbial Sensitivity Tests , Phenylethyl Alcohol/administration & dosage , Phenylethyl Alcohol/pharmacology , Phosphoproteins/chemistry , Plant Extracts/chemistry , /physiology , Skin Cream , Spike Glycoprotein, Coronavirus/chemistry , Vero Cells , Virus Inactivation/drug effects
2.
Sci Rep ; 11(1): 2418, 2021 01 28.
Article in English | MEDLINE | ID: covidwho-1054060

ABSTRACT

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection is currently a global pandemic, and there are limited laboratory studies targeting pathogen resistance. This study aimed to investigate the effect of selected disinfection products and methods on the inactivation of SARS-CoV-2 in the laboratory. We used quantitative suspension testing to evaluate the effectiveness of the disinfectant/method. Available chlorine of 250 mg/L, 500 mg/L, and 1000 mg/L required 20 min, 5 min, and 0.5 min to inactivate SARS-CoV-2, respectively. A 600-fold dilution of 17% concentration of di-N-decyl dimethyl ammonium bromide (283 mg/L) and the same concentration of di-N-decyl dimethyl ammonium chloride required only 0.5 min to inactivate the virus efficiently. At 30% concentration for 1 min and 40% and above for 0.5 min, ethanol could efficiently inactivate SARS-CoV-2. Heat takes approximately 30 min at 56 °C, 10 min above 70 °C, or 5 min above 90 °C to inactivate the virus. The chlorinated disinfectants, Di-N-decyl dimethyl ammonium bromide/chloride, ethanol, and heat could effectively inactivate SARS-CoV-2 in the laboratory test. The response of SARS-CoV-2 to disinfectants is very similar to that of SARS-CoV.


Subject(s)
Disinfectants/pharmacology , Disinfection/methods , Virus Inactivation/drug effects , /prevention & control , Chlorine/chemistry , Chlorine/pharmacology , Disinfectants/chemistry , Ethanol/chemistry , Ethanol/pharmacology , Humans , Pandemics/prevention & control , Quaternary Ammonium Compounds/chemistry , Quaternary Ammonium Compounds/pharmacology
3.
Viruses ; 13(1)2020 12 24.
Article in English | MEDLINE | ID: covidwho-1042658

ABSTRACT

The newly identified pathogenic human coronavirus, SARS-CoV-2, led to an atypical pneumonia-like severe acute respiratory syndrome (SARS) outbreak called coronavirus disease 2019 (abbreviated as COVID-19). Currently, nearly 77 million cases have been confirmed worldwide with the highest numbers of COVID-19 cases in the United States. Individuals are getting vaccinated with recently approved vaccines, which are highly protective in suppressing COVID-19 symptoms but there will be a long way before the majority of individuals get vaccinated. In the meantime, safety precautions and effective disease control strategies appear to be vital for preventing the virus spread in public places. Due to the longevity of the virus on smooth surfaces, photocatalytic properties of "self-disinfecting/cleaning" surfaces appear to be a promising tool to help guide disinfection policies for controlling SARS-CoV-2 spread in high-traffic areas such as hospitals, grocery stores, airports, schools, and stadiums. Here, we explored the photocatalytic properties of nanosized TiO2 (TNPs) as induced by the UV radiation, towards virus deactivation. Our preliminary results using a close genetic relative of SAR-CoV-2, HCoV-NL63, showed the virucidal efficacy of photoactive TNPs deposited on glass coverslips, as examined by quantitative RT-qPCR and virus infectivity assays. Efforts to extrapolate the underlying concepts described in this study to SARS-CoV-2 are currently underway.


Subject(s)
Disinfection/methods , Metal Nanoparticles/chemistry , /radiation effects , Titanium/pharmacology , Animals , Antiviral Agents/pharmacology , /virology , Chlorocebus aethiops , Disease Outbreaks/prevention & control , HEK293 Cells , Humans , Titanium/chemistry , Ultraviolet Rays , Vero Cells , Virus Inactivation/drug effects , Virus Inactivation/radiation effects
4.
ACS Appl Mater Interfaces ; 13(5): 5919-5928, 2021 Feb 10.
Article in English | MEDLINE | ID: covidwho-1042382

ABSTRACT

The ongoing COVID-19 pandemic has created a need for coatings that reduce infection from SARS-CoV-2 via surfaces. Such a coating could be used on common touch surfaces (e.g., door handles and railings) to reduce both disease transmission and fear of touching objects. Herein, we describe the design, fabrication, and testing of a cupric oxide anti-SARS-CoV-2 coating. Rapid loss of infectivity is an important design criterion, so a porous hydrophilic coating was created to allow rapid infiltration of aqueous solutions into the coating where diffusion distances to the cupric oxide surface are short and the surface area is large. The coating was deposited onto glass from a dispersion of cuprous oxide in ethanol and then thermally treated at 700 °C for 2 h to produce a CuO coating that is ≈30 µm thick. The heat treatment oxidized the cuprous oxide to cupric oxide and sintered the particles into a robust film. The SARS-CoV-2 infectivity from the CuO film was reduced by 99.8% in 30 min and 99.9% in 1 h compared to that from glass. The coating remained hydrophilic for at least 5 months, and there was no significant change in the cross-hatch test of robustness after exposure to 70% ethanol or 3 wt % bleach.


Subject(s)
/prevention & control , Copper/pharmacology , /physiology , Animals , Chlorocebus aethiops , Humans , Photoelectron Spectroscopy , Surface Properties , Vero Cells , Virus Inactivation/drug effects , X-Ray Diffraction
5.
Emerg Microbes Infect ; 10(1): 206-210, 2021 Dec.
Article in English | MEDLINE | ID: covidwho-1007445

ABSTRACT

The extremely rapid spread of the SARS-CoV-2 has already resulted in more than 1 million reported deaths of coronavirus disease 2019 (COVID-19). The ability of infectious particles to persist on environmental surfaces is potentially considered a factor for viral spreading. Therefore, limiting viral diffusion in public environments should be achieved with correct disinfection of objects, tissues, and clothes. This study proves how two widespread disinfection systems, short-wavelength ultraviolet light (UV-C) and ozone (O3), are active in vitro on different commonly used materials. The development of devices equipped with UV-C, or ozone generators, may prevent the virus from spreading in public places.


Subject(s)
/prevention & control , Disinfection/methods , Ozone/pharmacology , Ultraviolet Rays , Virus Inactivation/drug effects , Virus Inactivation/radiation effects , Humans , /isolation & purification , /radiation effects
6.
F1000Res ; 9: 674, 2020.
Article in English | MEDLINE | ID: covidwho-902997

ABSTRACT

Background: The ability to protect workers and healthcare professionals from infection by SARS-CoV-2, the virus that causes coronavirus disease 2019 (COVID-19), is of great concern. Hospitals, nursing homes and employers are adopting infection control strategies based on guidance from leading public health organizations such as the CDC, OSHA, FDA, and other government bodies. Certain hard surface disinfectants are effective against SARS-CoV-2 but are not suitable for use on skin or personal protective equipment (PPE) that comes into contact with skin. Furthermore, near-ubiquitous alcohol-based hand sanitizers are acceptable for use on skin, but they are not suitable for use on PPE. PPE, especially masks, are also commonly being used for longer durations than normal. There is a need for new products and techniques that can effectively disinfect PPE during wear time without having detrimental effects on surrounding skin. Clyraguard spray is a novel copper iodine complex designed to be used on non-critical PPE. Methods: In this study, the Clyraguard copper iodine complex was tested for its ability to inactivate SARS-CoV-2 in solution. Results: These data indicate the product to be effective in reducing SARS-CoV-2 titers in a time-dependent manner, with the virus being reduced below the detection limits within 30 minutes. Conclusions: These results suggest that Clyraguard may be an effective tool for mitigating cross-contamination of non-critical PPE that may come into contact with SARS-CoV-2.


Subject(s)
Betacoronavirus/drug effects , Copper/pharmacology , Disinfectants/pharmacology , Iodine/pharmacology , Virus Inactivation/drug effects , Coronavirus Infections , Humans , Pandemics , Pneumonia, Viral
7.
Viruses ; 12(11)2020 10 23.
Article in English | MEDLINE | ID: covidwho-895403

ABSTRACT

Critical to facilitating SARS-CoV-2 point-of-care (POC) testing is assurance that viruses present in specimens are inactivated onsite prior to processing. Here, we conducted experiments to determine the virucidal activity of commercially available Viral Transport Mediums (VTMs) to inactivate SARS-CoV-2. Independent testing methods for viral inactivation testing were applied, including a previously described World Health Organization (WHO) protocol, in addition to a buffer exchange method where the virus is physically separated from the VTM post exposure. The latter method enables sensitive detection of viral viability at higher viral titre when incubated with VTM. We demonstrate that VTM formulations, Primestore® Molecular Transport Medium (MTM) and COPAN eNAT™ completely inactivate high-titre SARS-CoV-2 virus (>1 × 107 copies/mL) and are compatible with POC processing. Furthermore, full viral inactivation was rapidly achieved in as little as 2 min of VTM exposure. We conclude that adding certain VTM formulations as a first step post specimen collection will render SARS-CoV-2 non-infectious for transport, or for further in-field POC molecular testing using rapid turnaround GeneXpert platforms or equivalent.


Subject(s)
Betacoronavirus/isolation & purification , Point-of-Care Testing , Specimen Handling , Virus Inactivation , Clinical Laboratory Techniques , Coronavirus Infections/diagnosis , Culture Media/analysis , Culture Media/pharmacology , Humans , Point-of-Care Testing/standards , Specimen Handling/methods , Specimen Handling/standards , Viral Load/drug effects , Virus Inactivation/drug effects
8.
J Virol Methods ; 287: 113996, 2021 01.
Article in English | MEDLINE | ID: covidwho-894099

ABSTRACT

BACKGROUND: Coronavirus disease 2019 (COVID-19) pandemic caused by infection with severe acute respiratory syndrome - coronavirus-2 (SARS-CoV-2) continues to affect many countries and large populations. Serologic assays for antibody detection aid patient diagnosis and seroepidemiologic investigations. METHODS: An indirect IgG ELISA was developed indigenously using ß-propiolactone (BPL) inactivated SARS-CoV-2. This assay was used for screening 200 healthy donor sera collected prior to COVID-19 emergence (2017-2019), 185 serum/plasma samples of confirmed COVID-19 patients (n = 137) and 57 samples of viral RNA positive asymptomatic contacts (n = 51). The IgG response was studied in relation to duration and severity of illness. RESULTS: The ELISA demonstrated 97 % specificity and IgG detection in >50 %, 80 %, 93.8 % and 100 % of the patients respectively during the first, second, third and fourth week of illness. IgG detection rate was higher in patients with severe disease (SD, 90.9 %) than those with mild disease (MD, 68.8 %) during the second week of illness (P = 0.027). IgG seropositivity among asymptomatic contacts was 64.7 %. IgG ELISA absorbance values were higher in SD than MD patients during the first 2 weeks of illness (P < 0.05). No significant difference was observed between the absorbance values of asymptomatic subjects and MD patients (P = 0.94). CONCLUSION: The BPL inactivated virus-based ELISA could detect IgG antibodies early and in a significant proportion of COVID-19 patients suggesting its potential utility as a supplement to the currently used viral RNA detection tests in patient diagnosis and contact screening algorithms.


Subject(s)
/methods , Immunoglobulin G/blood , Propiolactone/pharmacology , /immunology , Antibodies, Viral/blood , /pathology , Enzyme-Linked Immunosorbent Assay , Humans , Sensitivity and Specificity , Seroepidemiologic Studies , Virus Inactivation/drug effects
9.
J Clin Microbiol ; 58(11)2020 10 21.
Article in English | MEDLINE | ID: covidwho-729361

ABSTRACT

The COVID-19 pandemic has necessitated a multifaceted rapid response by the scientific community, bringing researchers, health officials, and industry together to address the ongoing public health emergency. To meet this challenge, participants need an informed approach for working safely with the etiological agent, the novel human coronavirus SARS-CoV-2. Work with infectious SARS-CoV-2 is currently restricted to high-containment laboratories, but material can be handled at a lower containment level after inactivation. Given the wide array of inactivation reagents that are being used in laboratories during this pandemic, it is vital that their effectiveness is thoroughly investigated. Here, we evaluated a total of 23 commercial reagents designed for clinical sample transportation, nucleic acid extraction, and virus inactivation for their ability to inactivate SARS-CoV-2, as well as seven other common chemicals, including detergents and fixatives. As part of this study, we have also tested five filtration matrices for their effectiveness at removing the cytotoxic elements of each reagent, permitting accurate determination of levels of infectious virus remaining following treatment. In addition to providing critical data informing inactivation methods and risk assessments for diagnostic and research laboratories working with SARS-CoV-2, these data provide a framework for other laboratories to validate their inactivation processes and to guide similar studies for other pathogens.


Subject(s)
Betacoronavirus/drug effects , Indicators and Reagents/pharmacology , Virus Inactivation/drug effects , Animals , Betacoronavirus/isolation & purification , Cell Survival/drug effects , Chlorocebus aethiops , Clinical Laboratory Techniques , Coronavirus Infections/diagnosis , Coronavirus Infections/virology , Filtration/instrumentation , Humans , Pandemics , Pneumonia, Viral/diagnosis , Pneumonia, Viral/virology , Vero Cells
10.
Biochem Biophys Res Commun ; 530(1): 1-3, 2020 09 10.
Article in English | MEDLINE | ID: covidwho-641528

ABSTRACT

Alcohol-based disinfectant shortage is a serious concern in the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. Acidic electrolyzed water (EW) with a high concentration of free available chlorine (FAC) shows strong antimicrobial activity against bacteria, fungi, and viruses. Here, we assessed the SARS-CoV-2-inactivating efficacy of acidic EW for use as an alternative disinfectant. The quick virucidal effect of acidic EW depended on the concentrations of contained-FAC. The effect completely disappeared in acidic EW in which FAC was lost owing to long-time storage after generation. In addition, the virucidal activity increased proportionately with the volume of acidic EW mixed with the virus solution when the FAC concentration in EW was same. These findings suggest that the virucidal activity of acidic EW against SARS-CoV-2 depends on the amount of FAC contacting the virus.


Subject(s)
Betacoronavirus/drug effects , Chlorine/pharmacology , Disinfectants/pharmacology , Disinfection/methods , Virus Inactivation/drug effects , Acids/chemistry , Acids/pharmacology , Antiviral Agents/chemistry , Antiviral Agents/pharmacology , Betacoronavirus/physiology , Chlorine/chemistry , Coronavirus Infections/prevention & control , Coronavirus Infections/virology , Disinfectants/chemistry , Electrolysis/methods , Humans , Pandemics/prevention & control , Pneumonia, Viral/prevention & control , Pneumonia, Viral/virology , Water/chemistry , Water/pharmacology
11.
Viruses ; 12(6)2020 06 08.
Article in English | MEDLINE | ID: covidwho-574875

ABSTRACT

Clinical samples collected in coronavirus disease 19 (COVID-19), patients are commonly manipulated in biosafety level 2 laboratories for molecular diagnostic purposes. Here, we tested French norm NF-EN-14476+A2 derived from European standard EN-14885 to assess the risk of manipulating infectious viruses prior to RNA extraction. SARS-CoV-2 cell-culture supernatant and nasopharyngeal samples (virus-spiked samples and clinical samples collected in COVID-19 patients) were used to measure the reduction of infectivity after 10 minute contact with lysis buffer containing various detergents and chaotropic agents. A total of thirteen protocols were evaluated. Two commercially available formulations showed the ability to reduce infectivity by at least 6 log 10, whereas others proved less effective.


Subject(s)
Betacoronavirus/drug effects , Coronavirus Infections/virology , Pneumonia, Viral/virology , Virus Inactivation/drug effects , Animals , Betacoronavirus/genetics , Betacoronavirus/isolation & purification , Betacoronavirus/physiology , Cell Culture Techniques/methods , Chlorocebus aethiops , Containment of Biohazards/methods , Containment of Biohazards/standards , Humans , Nasopharynx/virology , Pandemics , RNA, Viral/isolation & purification , Specimen Handling/methods , Vero Cells , Viral Load/methods
12.
J Clin Microbiol ; 58(8)2020 Jul 23.
Article in English | MEDLINE | ID: covidwho-421116

ABSTRACT

The outbreak of coronavirus disease 2019 (COVID-19) has spread across the world and was characterized as a pandemic. To protect medical laboratory personnel from infection, most laboratories inactivate the virus causing COVID-19, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), in clinical samples before testing. However, the effect of inactivation on the detection results remains unknown. Here, we used a digital PCR assay to determine the absolute SARS-CoV-2 RNA copy number in 63 nasopharyngeal swab samples and assess the effect of inactivation methods on viral RNA copy number. Viral inactivation was performed by three different methods: (i) incubation with the TRIzol LS reagent for 10 min at room temperature, (ii) heating in a water bath at 56°C for 30 min, and (iii) high-temperature treatment, including autoclaving at 121°C for 20 min, boiling at 100°C for 20 min, and heating at 80°C for 20 min. Compared to the amount of RNA in the original sample, TRIzol treatment destroyed 47.54% of the nucleocapsid protein (N) gene and 39.85% of open reading frame (ORF) 1ab. For samples treated at 56°C for 30 min, the copy number of the N gene and ORF 1ab was reduced by 48.55% and 56.40%, respectively. The viral RNA copy number dropped by 50 to 66% after heating at 80°C for 20 min. Nearly no viral RNA was detected after autoclaving at 121°C or boiling at 100°C for 20 min. These results indicate that inactivation reduced the quantity of detectable viral RNA and may cause false-negative results, especially in weakly positive cases. Thus, use of the TRIzol reagent rather than heat inactivation is recommended for sample inactivation, as the TRIzol reagent had the least effect on the RNA copy number among the tested methods.


Subject(s)
Betacoronavirus/drug effects , Betacoronavirus/radiation effects , Disinfection/methods , RNA, Viral/analysis , Specimen Handling/methods , Virus Inactivation/drug effects , Virus Inactivation/radiation effects , Adolescent , Adult , Aged , Aged, 80 and over , Disinfectants , Female , Gene Dosage , Hot Temperature , Humans , Male , Middle Aged , Polymerase Chain Reaction , RNA, Viral/genetics , Young Adult
13.
Clin Chem ; 66(6): 794-801, 2020 06 01.
Article in English | MEDLINE | ID: covidwho-30705

ABSTRACT

BACKGROUND: Coronavirus disease-2019 (COVID-19) has spread widely throughout the world since the end of 2019. Nucleic acid testing (NAT) has played an important role in patient diagnosis and management of COVID-19. In some circumstances, thermal inactivation at 56°C has been recommended to inactivate severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) before NAT. However, this procedure could theoretically disrupt nucleic acid integrity of this single-stranded RNA virus and cause false negatives in real-time polymerase chain reaction (RT-PCR) tests. METHODS: We investigated whether thermal inactivation could affect the results of viral NAT. We examined the effects of thermal inactivation on the quantitative RT-PCR results of SARS-CoV-2, particularly with regard to the rates of false-negative results for specimens carrying low viral loads. We additionally investigated the effects of different specimen types, sample preservation times, and a chemical inactivation approach on NAT. RESULTS: Our study showed increased Ct values in specimens from diagnosed COVID-19 patients in RT-PCR tests after thermal incubation. Moreover, about half of the weak-positive samples (7 of 15 samples, 46.7%) were RT-PCR negative after heat inactivation in at least one parallel testing. The use of guanidinium-based lysis for preservation of these specimens had a smaller impact on RT-PCR results with fewer false negatives (2 of 15 samples, 13.3%) and significantly less increase in Ct values than heat inactivation. CONCLUSION: Thermal inactivation adversely affected the efficiency of RT-PCR for SARS-CoV-2 detection. Given the limited applicability associated with chemical inactivators, other approaches to ensure the overall protection of laboratory personnel need consideration.


Subject(s)
Betacoronavirus/chemistry , Coronavirus Infections/diagnosis , Hot Temperature , Pneumonia, Viral/diagnosis , RNA, Viral/analysis , Viral Load , Clinical Laboratory Techniques/methods , False Negative Reactions , Feces/virology , Guanidine/chemistry , Humans , Pandemics , Real-Time Polymerase Chain Reaction/methods , Real-Time Polymerase Chain Reaction/statistics & numerical data , Specimen Handling/methods , Sputum/virology , Time Factors , Virus Inactivation/drug effects
14.
Arch Med Res ; 51(3): 282-286, 2020 04.
Article in English | MEDLINE | ID: covidwho-23547

ABSTRACT

SARS-CoV-2, SARS and MERS are all enveloped viruses that can cause acute respiratory syndrome. Arachidonic acid (AA) and other unsaturated fatty acids (especially eicosapentaenoic acd, EPA and docosahexaenoic acid DHA) are known to inactivate enveloped viruses and inhibit proliferation of various microbial organisms. The pro-inflammatory metabolites of AA and EPA such as prostaglandins, leukotrienes and thromboxanes induce inflammation whereas lipoxins, resolvins, protectins and maresins derived from AA, EPA and DHA not only suppress inflammation but also enhance would healing and augment phagocytosis of macrophages and other immunocytes and decrease microbial load. In view of these actions, it is suggested that AA and other unsaturated fatty acids and their metabolites may serve as endogenous anti-viral compounds and their deficiency may render humans susceptible to SARS-CoV-2, SARS and MERS and other similar viruses' infections. Hence, oral or intravenous administration of AA and other unsaturated fatty acids may aid in enhancing resistance and recovery from SARS-CoV-2, SARS and MERS infections.


Subject(s)
Betacoronavirus/drug effects , Coronavirus Infections/drug therapy , Coronavirus Infections/virology , Fatty Acids, Unsaturated/therapeutic use , Pneumonia, Viral/drug therapy , Pneumonia, Viral/virology , Virus Inactivation/drug effects , Coronavirus Infections/epidemiology , Coronavirus Infections/pathology , Fatty Acids, Unsaturated/pharmacology , Humans , Models, Molecular , Pandemics , Pneumonia, Viral/epidemiology , Pneumonia, Viral/pathology
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