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1.
Vet Pathol ; 59(4): 556-564, 2022 07.
Article in English | MEDLINE | ID: covidwho-1785003

ABSTRACT

The COVID-19 pandemic has highlighted the critical role that animal models play in elucidating the pathogenesis of emerging diseases and rapidly analyzing potential medical countermeasures. Relevant pathologic outcomes are paramount in evaluating preclinical models and therapeutic outcomes and require careful advance planning. While there are numerous guidelines for attaining high-quality pathology specimens in routine animal studies, preclinical studies using coronaviruses are often conducted under biosafety level-3 (BSL3) conditions, which pose unique challenges and technical limitations. In such settings, rather than foregoing pathologic outcomes because of the inherent constraints of high-containment laboratory protocols, modifications can be made to conventional best practices of specimen collection. Particularly for those unfamiliar with working in a high-containment laboratory, the authors describe the logistics of conducting such work, focusing on animal experiments in BSL3 conditions. To promote scientific rigor and reproducibility and maximize the value of animal use, the authors provide specific points to be considered before, during, and following a high-containment animal study. The authors provide procedural modifications for attaining good quality pathologic assessment of the mouse lung, central nervous system, and blood specimens under high-containment conditions while being conscientious to maximize animal use for other concurrent assays.


Subject(s)
COVID-19 , Containment of Biohazards , Laboratories , Specimen Handling , Animals , COVID-19/veterinary , Containment of Biohazards/standards , Laboratories/organization & administration , Mice , Reproducibility of Results , SARS-CoV-2 , Specimen Handling/methods , Specimen Handling/veterinary
2.
Viruses ; 12(6)2020 06 08.
Article in English | MEDLINE | ID: covidwho-1726020

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 , COVID-19 , Cell Culture Techniques/methods , Chlorocebus aethiops , Containment of Biohazards/methods , Containment of Biohazards/standards , Humans , Nasopharynx/virology , Pandemics , RNA, Viral/isolation & purification , SARS-CoV-2 , Specimen Handling/methods , Vero Cells , Viral Load/methods
3.
J Infect Dev Ctries ; 15(12): 1833-1837, 2021 12 31.
Article in English | MEDLINE | ID: covidwho-1633510

ABSTRACT

At the beginning of the coronavirus disease 2019 (COVID-19) pandemic in Bangladesh, there was a scarcity of ideal biocontainment facilities to detect the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a risk group of 3 organisms. Molecular detection of SARS-CoV-2 must be performed in a BSL-2 laboratory with BSL-3-equivalent infection prevention and control practices. Establishing these facilities within a short timeframe proved to be an enormous challenge, including locating a remote space distant from the university campus to establish a laboratory, motivating the laboratory staff to work with a novel pathogen without any prior experience, allocation of funds for essential equipment and accessories, and arrangement of a safe waste management system for environmental hazard reduction. This report also highlights several limitations, such as the facility's architectural design that did not follow the biosafety guidelines, lack of continuous flow of funds, and an inadequate number of laboratory personnel. This article describes various efforts taken to overcome the challenges during the establishment of this facility that may be adopted to create similar facilities in other regions of the country. Establishing a BSL-2 laboratory with BSL-3-equivalent infection prevention and control practices will aid in the early detection of a large number of cases, thereby isolating persons with COVID-19, limiting the transmission of SARS-CoV-2, and promoting a robust public health response to contain the pandemic.


Subject(s)
COVID-19 Testing/methods , COVID-19/diagnosis , Containment of Biohazards/standards , Facility Design and Construction/methods , Laboratories/standards , Bangladesh/epidemiology , COVID-19/epidemiology , Humans , Pandemics/prevention & control , SARS-CoV-2
4.
J Virol ; 96(3): e0183721, 2022 02 09.
Article in English | MEDLINE | ID: covidwho-1546443

ABSTRACT

Research activities with infectious severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are currently permitted only under biosafety level 3 (BSL3) containment. Here, we report the development of a single-cycle infectious SARS-CoV-2 virus replicon particle (VRP) system with a luciferase and green fluorescent protein (GFP) dual reporter that can be safely handled in BSL2 laboratories to study SARS-CoV-2 biology. The spike (S) gene of SARS-CoV-2 encodes the envelope glycoprotein, which is essential for mediating infection of new host cells. Through deletion and replacement of this essential S gene with a luciferase and GFP dual reporter, we have generated a conditional SARS-CoV-2 mutant (ΔS-VRP) that produces infectious particles only in cells expressing a viral envelope glycoprotein of choice. Interestingly, we observed more efficient production of infectious particles in cells expressing vesicular stomatitis virus (VSV) glycoprotein G [ΔS-VRP(G)] than in cells expressing other viral glycoproteins, including S. We confirmed that infection from ΔS-VRP(G) is limited to a single round and can be neutralized by anti-VSV serum. In our studies with ΔS-VRP(G), we observed robust expression of both luciferase and GFP reporters in various human and murine cell types, demonstrating that a broad variety of cells can support intracellular replication of SARS-CoV-2. In addition, treatment of ΔS-VRP(G)-infected cells with either of the anti-CoV drugs remdesivir (nucleoside analog) and GC376 (CoV 3CL protease inhibitor) resulted in a robust decrease in both luciferase and GFP expression in a drug dose- and cell-type-dependent manner. Taken together, our findings show that we have developed a single-cycle infectious SARS-CoV-2 VRP system that serves as a versatile platform to study SARS-CoV-2 intracellular biology and to perform high-throughput screening of antiviral drugs under BSL2 containment. IMPORTANCE Due to the highly contagious nature of SARS-CoV-2 and the lack of immunity in the human population, research on SARS-CoV-2 has been restricted to biosafety level 3 laboratories. This has greatly limited participation of the broader scientific community in SARS-CoV-2 research and thus has hindered the development of vaccines and antiviral drugs. By deleting the essential spike gene in the viral genome, we have developed a conditional mutant of SARS-CoV-2 with luciferase and fluorescent reporters, which can be safely used under biosafety level 2 conditions. Our single-cycle infectious SARS-CoV-2 virus replicon system can serve as a versatile platform to study SARS-CoV-2 intracellular biology and to perform high-throughput screening of antiviral drugs under BSL2 containment.


Subject(s)
Genetic Engineering , Recombination, Genetic , Replicon , SARS-CoV-2/genetics , COVID-19/virology , Cell Culture Techniques , Cell Line , Containment of Biohazards/standards , Genes, Reporter , Humans , Laboratories/standards , Viral Proteins/genetics , Virus Replication
10.
Cytometry A ; 99(1): 68-80, 2021 01.
Article in English | MEDLINE | ID: covidwho-1086342

ABSTRACT

Biosafety has always been an important aspect of daily work in any research institution, particularly for cytometry Shared Resources Laboratories (SRLs). SRLs are common-use spaces that facilitate the sharing of knowledge, expertise, and ideas. This sharing inescapably involves contact and interaction of all those within this working environment on a daily basis. The current pandemic caused by SARS-CoV-2 has prompted the re-evaluation of many policies governing the operations of SRLs. Here we identify and review the unique challenges SRLs face in maintaining biosafety standards, highlighting the potential risks associated with not only cytometry instrumentation and samples, but also the people working with them. We propose possible solutions to safety issues raised by the COVID-19 pandemic and provide tools for facilities to adapt to evolving guidelines and future challenges.


Subject(s)
COVID-19/epidemiology , Containment of Biohazards/trends , Laboratories/trends , COVID-19/prevention & control , COVID-19/transmission , Containment of Biohazards/standards , Flow Cytometry , Humans , Laboratories/standards , Risk Assessment/standards , Risk Assessment/trends
11.
Pathologica ; 112(4): 174-177, 2020 Dec.
Article in English | MEDLINE | ID: covidwho-1022379

ABSTRACT

Up to now, Italy is one of the European centers with the most active Coronavirus cases with 233,836 positive cases and 33,601 total deaths as of June 3rd. During this pandemic and dramatic emergency, Italian hospitals had also to face neoplastic pathologies, that still afflict the Italian population, requiring urgent surgical and oncological treatment. In our Cancer Center Hospital, the high volume of surgical procedures have demanded an equally high volume of intraoperative pathological examinations, but also posed an additional major challenge for the safety of the staff involved. The current commentary reports our experience in the past two months (since March 9th) for a total of 1271 frozen exams from 893 suspect COVID-19 patients (31 confirmed).


Subject(s)
COVID-19 , Containment of Biohazards/standards , Intraoperative Care/standards , Pandemics , Pathology/standards , COVID-19/epidemiology , Humans , Intraoperative Care/statistics & numerical data , Italy/epidemiology , Middle Aged , Pathology/statistics & numerical data
12.
Cytometry A ; 99(1): 68-80, 2021 01.
Article in English | MEDLINE | ID: covidwho-1009048

ABSTRACT

Biosafety has always been an important aspect of daily work in any research institution, particularly for cytometry Shared Resources Laboratories (SRLs). SRLs are common-use spaces that facilitate the sharing of knowledge, expertise, and ideas. This sharing inescapably involves contact and interaction of all those within this working environment on a daily basis. The current pandemic caused by SARS-CoV-2 has prompted the re-evaluation of many policies governing the operations of SRLs. Here we identify and review the unique challenges SRLs face in maintaining biosafety standards, highlighting the potential risks associated with not only cytometry instrumentation and samples, but also the people working with them. We propose possible solutions to safety issues raised by the COVID-19 pandemic and provide tools for facilities to adapt to evolving guidelines and future challenges.


Subject(s)
COVID-19/epidemiology , Containment of Biohazards/trends , Laboratories/trends , COVID-19/prevention & control , COVID-19/transmission , Containment of Biohazards/standards , Flow Cytometry , Humans , Laboratories/standards , Risk Assessment/standards , Risk Assessment/trends
13.
Clin Chim Acta ; 512: 58-62, 2021 Jan.
Article in English | MEDLINE | ID: covidwho-956958

ABSTRACT

OBJECTIVE: It is unclear if implementation of biosafety action plans in response to the COVID-19 pandemic has affected laboratory quality metrics. METHODS: This retrospective study used quality data, including turnaround time (TAT) and number/type of unacceptable specimens from a stat laboratory supporting an outpatient medical clinic serving predominantly elderly cancer patients. Four months of data from the height of the COVID-19 pandemic (March-June 2020) were compared to the same months in 2019. RESULTS: March-May 2020 test volumes were decreased compared to 2019. June 2020 test volume was slightly increased compared to 2019. TATs in 2020 were similar/ slightly improved compared to the same months in 2019, due to shortened collect to receive and receive to verify TATs. The number and types of unacceptable specimens were similar in 2020 and 2019. CONCLUSIONS: Despite the challenges to the system caused by the pandemic, laboratory quality metrics were maintained.


Subject(s)
COVID-19 , Laboratories/statistics & numerical data , Aged , COVID-19/epidemiology , Containment of Biohazards/standards , Humans , Laboratories/standards , Neoplasms , Pandemics , Retrospective Studies , Specimen Handling/standards , Specimen Handling/statistics & numerical data , Time Factors
14.
Zhonghua Liu Xing Bing Xue Za Zhi ; 41(9): 1378-1380, 2020 Sep 10.
Article in Chinese | MEDLINE | ID: covidwho-881373

ABSTRACT

Biosafety is an important guarantee of the new coronavirus laboratory test. The accident treatment of sample overflow and sprinkle is a necessary part of the emergency plan for testing activities. Beijing Preventive Medicine Association coordinated biosafety experts of COVID-19 laboratories from Beijing CDC, to write up "The standard for handling of accidents of corona virus disease 2019 sample (T/BPMA 0005-2020)" . The group standard was based on the guidelines of China and WHO, and combined with the practical experience of COVID-19 epidemic and the principle of "scientific, normative, applicable and feasible" . Through all kinds of risk Assessment, it included the spillover of samples caused by the packing of COVID-19 (highly pathogenic) samples, the overflow and sprinkle in the laboratory during the detection operation, and the spillage accident occurred during the transfer of samples in the same building. The standard could guide and standardize the handling methods of accidental overflow and sprinkle that may occur in the SARS-CoV-2 testing laboratories in the city.


Subject(s)
Biohazard Release , Clinical Laboratory Techniques , Containment of Biohazards/standards , Beijing , COVID-19 Testing , Coronavirus Infections/diagnosis , Humans
16.
Rev. enferm. UERJ ; 28: e50360, jan.-dez. 2020.
Article in English, Portuguese | WHO COVID, LILACS (Americas) | ID: covidwho-831603

ABSTRACT

Objetivo: descrever as recomendações sobre o uso racional e seguro dos equipamentos de proteção individual (EPI) no transcorrer da cadeia assistencial de pessoas com suspeita ou confirmação de contaminação pelo novo coronavírus. Conteúdo: o novo coronavírus é responsável pela doença Covid-19, e dentre as pessoas com maior risco de desenvolver a infecção estão os trabalhadores de saúde, devido ao contato muito próximo a pacientes. Desse modo, a utilização de EPI é recomendação prioritária a estes trabalhadores. Todavia, em função do desabastecimento internacional e nacional relacionado a estes equipamentos, o uso racional é fundamental a fim de evitar que o impacto do desabastecimento seja ainda maior. Conclusão: o uso de EPI é indispensável aos trabalhadores de saúde durante a pandemia de Covid-19, contudo, é imprescindível coordenar a cadeia de fornecimento destes insumos, implementar estratégias que minimizem a necessidade de EPI e garantir o uso de maneira adequada.


Objective: to describe the recommendations on the rational, safe use of personal protective equipment (PPE) throughout the chain of care for people with suspected or confirmed contamination by the new coronavirus. Content: the new coronavirus is responsible for the disease Covid-19, and among those at high risk of infection are health workers in very close contact with patients. It is thus a priority recommendation for these workers to use PPE. However, international and national shortages of this equipment make rational use essential in order to prevent even greater impact from these shortages. Conclusion: it is essential that health workers use PPE during the Covid-19 pandemic, but it is also essential to coordinate the supply chain for these inputs, implement strategies that minimize the need for PPE and ensure proper use.


Objetivo: describir las recomendaciones sobre el uso racional y seguro del equipo de protección personal (EPP) en toda la cadena de atención para las personas con sospecha o confirmación de contaminación por el nuevo coronavirus. Contenido: el nuevo coronavirus es responsable de la enfermedad de Covid-19, y entre aquellos con alto riesgo de infección se encuentran los trabajadores de la salud en contacto muy cercano con los pacientes. Por lo tanto, es una recomendación prioritaria para estos trabajadores usar EPP. Sin embargo, la escasez internacional y nacional de este equipo hace que el uso racional sea esencial para evitar un impacto aún mayor de esta escasez. Conclusión: es esencial que los trabajadores de la salud usen EPP durante la pandemia de Covid-19, pero también es esencial coordinar la cadena de suministro para estos insumos, implementar estrategias que minimicen la necesidad de EPP y garantizar un uso adecuado.


Subject(s)
Coronavirus Infections/prevention & control , Personal Protective Equipment/supply & distribution , Betacoronavirus , Masks/supply & distribution , Occupational Risks , Containment of Biohazards/standards , Personal Protective Equipment/standards , Masks/standards
17.
Rev. enferm. UERJ ; 28: e50721, jan.-dez. 2020.
Article in English, Portuguese | WHO COVID, LILACS (Americas) | ID: covidwho-828134

ABSTRACT

Objetivo: apresentar atualizações para a ressuscitação cardiopulmonar em pacientes suspeitos e confirmados com COVID-19. Método: revisão compreensiva da literatura, com síntese narrativa das evidências de diretrizes e recomendações da Organização Mundial de Saúde, Associação de Medicina Intensiva Brasileira, American Heart Association, Resuscitation Council UK, American College of Surgions Committee on Trauma e National Association of Emergency Medical Technicians. Resultados: as principais atualizações trazem informações sobre especificidades das manobras de ressuscitação cardiopulmonar; preparação do ambiente, recursos humanos e materiais, reconhecimento da parada cardiorrespiratória e ações iniciais; estratégias de ventilação e acesso invasivo da via aérea; ajustes do ventilador mecânico e manobras de ressuscitação cardiopulmonar em pacientes pronados. Considerações finais: profissionais de saúde envolvidos no atendimento à parada cardiorrespiratória de pacientes suspeitos e/ou confirmados com COVID-19 podem encontrar inúmeros desafios, portanto devem seguir com rigor o protocolo estabelecido para maximizar a efetividade das manobras de ressuscitação e minimizar o risco de contágio pelo vírus e sua disseminação.


Objective: to present updates for cardiopulmonary resuscitation in suspected and confirmed patients with COVID-19. Method: comprehensive literature review with narrative synthesis of the evidence of guidelines and recommendations from World Health Organization, Associação de Medicina Intensiva Brasileira, American Heart Association, Resuscitation Council UK, American College of Surgions Committee on Trauma and National Association of Emergency Medical Technicians. Results: the main updates bring information about the specifics of cardiopulmonary resuscitation maneuvers; preparation of the environment and human and material resources, recognition of cardiorespiratory arrest and initial actions; ventilation and invasive airway access strategies; mechanical ventilator adjustments and cardiopulmonary resuscitation maneuvers in patients in the prone position. Final considerations: health professionals involved in the care of cardiorespiratory arrest of suspected and/or confirmed patients with COVID-19 can face numerous challenges, so they must strictly follow the protocol established to maximize the effectiveness of resuscitation maneuvers and minimize the risk of contagion by the virus and its spread.


Objetivo: apresentar actualizaciones para la reanimación cardiopulmonar en pacientes sospechos os y confirmados con COVID-19. Método: revisión exhaustiva de la literatura con síntesis narrativa de la evidencia de guías y recomendaciones de la Organización Mundial de la Salud, Associação de Medicina Intensiva Brasileira, American Heart Association, Resuscitation Council UK, American College of Surgions Committee on Trauma and National Association of Emergency Medical Technicians. Resultados: las principales actualizaciones aportan información sobre los detalles de las maniobras de reanimación cardiopulmonar; preparación del medio ambiente y recursos humanos y materiales, reconocimiento de paro cardiorrespiratorio y acciones iniciales; estrategias de ventilación y acceso invasivo a las vías aéreas; ajustes del ventilador mecánico y maniobras de reanimación cardiopulmonar en pacientes en decúbito prono. Consideraciones finales: los profesionales de la salud involucrados en la atención del paro cardiorrespiratorio de pacientes sospechosos y/o confirmados con COVID-19 pueden enfrentar numerosos desafíos, por lo que deben seguir estrictamente el protocolo establecido para maximizar la efectividad de las maniobras de reanimación y minimizar el riesgo de contagio por el virus y supropagación.


Subject(s)
Humans , Male , Female , Cardiopulmonary Resuscitation/standards , Coronavirus Infections/complications , Betacoronavirus , Heart Arrest/etiology , Respiration, Artificial/methods , Clinical Protocols/standards , Cardiopulmonary Resuscitation/methods , Containment of Biohazards/standards , Heart Arrest/rehabilitation , Heart Massage/methods , Nursing, Team/standards
18.
Audiol., Commun. res ; 25: e2369, 2020.
Article in Portuguese | WHO COVID, LILACS (Americas) | ID: covidwho-818626

ABSTRACT

RESUMO A doença de coronavírus (COVID-19) é causada pela síndrome respiratória aguda grave coronavírus 2 (SARS-CoV-2). O vírus é transmitido, principalmente, por gotículas, espirros e aerossóis e pode ser transmitido mesmo entre pacientes assintomáticos, havendo risco de contágio durante os procedimentos do audiologista, que utiliza e reutiliza equipamentos clínicos em uma ampla variedade de pacientes. Este artigo teve como objetivo descrever as etapas que podem ser adotadas pelos audiologistas para diminuir o risco de contaminação cruzada na prática clínica, durante a pandemia de SARS-CoV-2. Recomenda-se, portanto, a esses profissionais, o uso de equipamentos de proteção individual, incluindo respiradores N95, luvas de procedimento, protetores para calçados descartáveis, protetores faciais ou óculos de segurança, gorros e aventais descartáveis, além de seguir, rigorosamente, os protocolos de biossegurança durante os cuidados audiológicos.


ABSTRACT Coronavirus disease (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-COV2). This virus is transmitted mainly by droplets, sneezes and aerosols and can be transmitted even among asymptomatic patients, so there is a risk of transmission during the audiologist's procedures which, in addition, use and reuse clinical equipment in a wide variety of patients. This article aims to describe the steps that can be taken by the audiologist in order to decrease the risk of cross-contamination in clinical practice during the SARS-CoV-2 pandemic in Brazil. During the COVID-19 pandemic, audiologists are recommended to use personal protective equipment including N95 respirators, clinical gloves, disposable shoe covers, face shields or safety glasses, hair covers and disposable aprons, in addition to strictly following biosafety protocols during audiological care.


Subject(s)
Health Personnel , Containment of Biohazards/standards , Personal Protective Equipment , COVID-19/prevention & control , COVID-19/epidemiology , Brazil/epidemiology , Audiology , Speech, Language and Hearing Sciences , SARS-CoV-2
20.
Viruses ; 12(7)2020 07 07.
Article in English | MEDLINE | ID: covidwho-639283

ABSTRACT

Standard precautions to minimize the risk of SARS-CoV-2 transmission implies that infected cell cultures and clinical specimens may undergo some sort of inactivation to reduce or abolish infectivity. We evaluated three heat inactivation protocols (56 °C-30 min, 60 °C-60 min and 92 °C-15 min) on SARS-CoV-2 using (i) infected cell culture supernatant, (ii) virus-spiked human sera (iii) and nasopharyngeal samples according to the recommendations of the European norm NF EN 14476-A2. Regardless of the protocol and the type of samples, a 4 Log10 TCID50 reduction was observed. However, samples containing viral loads > 6 Log10 TCID50 were still infectious after 56 °C-30 min and 60 °C-60 min, although infectivity was < 10 TCID50. The protocols 56 °C-30 min and 60 °C-60 min had little influence on the RNA copies detection, whereas 92 °C-15 min drastically reduced the limit of detection, which suggests that this protocol should be avoided for inactivation ahead of molecular diagnostics. Lastly, 56 °C-30 min treatment of serum specimens had a negligible influence on the results of IgG detection using a commercial ELISA test, whereas a drastic decrease in neutralizing titers was observed.


Subject(s)
Betacoronavirus , Containment of Biohazards/methods , Coronavirus Infections/virology , Pneumonia, Viral/virology , Serologic Tests/methods , Virus Inactivation , Antibodies, Neutralizing/immunology , Betacoronavirus/immunology , COVID-19 , Containment of Biohazards/standards , Coronavirus Infections/diagnosis , Coronavirus Infections/prevention & control , Enzyme-Linked Immunosorbent Assay , Hot Temperature , Humans , Neutralization Tests , Pandemics/prevention & control , Pneumonia, Viral/diagnosis , Pneumonia, Viral/prevention & control , SARS-CoV-2 , Serologic Tests/standards
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