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1.
J Infect Chemother ; 27(1): 117-119, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32994136

RESUMO

The novel coronavirus disease 2019 (COVID-19) is diagnosed by positive result of reverse transcription polymerase chain reaction (RT-PCR) for the novel coronavirus. We concluded that cycle threshold value (Ct-value) of real-time RT-PCR (rRT-PCR) assay could decrease as patients recover. Results of rRT-PCR assay could remain positive among asymptomatic patients for longer than 2 weeks. The discharge criteria of COVID-19 patients using a negative result of rRT-PCR should be reconsidered.


Assuntos
Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Adolescente , Adulto , Idoso de 80 Anos ou mais , Doenças Assintomáticas , Betacoronavirus/isolamento & purificação , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/virologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Nasofaringe/virologia , Pandemias , Alta do Paciente , Pneumonia Viral/virologia , Índice de Gravidade de Doença , Carga Viral , Adulto Jovem
2.
J Infect Chemother ; 27(1): 126-129, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33060046

RESUMO

Considering the issues of shortage of medical resources and the invasiveness and infection risk involved in the collection of nasopharyngeal swab specimens, there is a need for an effective alternative test specimen for SARS-CoV-2 RNA detection. Here, we investigated suitability of saliva as a non-invasively obtained specimen for molecular detection of SARS-CoV-2 RNA in Japanese patients with COVID-19. In total, 28 paired clinical specimens of saliva and nasopharyngeal swabs were collected from 12 patients at various time points after symptom onset. Each specimen was assayed using reverse transcription real-time polymerase chain reaction (rRT-PCR) on the BD MAX open system using primers and probes targeting the N-gene. The saliva and nasopharyngeal swab specimens showed 19 and 15 positive results, respectively. No invalid (PCR inhibition) result was observed for any specimen. The qualitative results of each specimen obtained in the period immediately after symptom onset were similar. Three convalescent patients presented saliva-positive results, whereas their nasopharyngeal swabs were negative at four different time points, suggesting that saliva may be superior to nasopharyngeal swabs in terms of obtaining stable assay result of SARS-CoV-2. In conclusion, our results suggest that saliva can potentially serve as an alternative to nasopharyngeal swabs as a specimen for SARS-CoV-2 rRT-PCR. As saliva can be collected by patients themselves, it may be an effective way to overcome the shortage of personal protective equipment and specimen sampling tools.


Assuntos
Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/diagnóstico , Nasofaringe/virologia , Pneumonia Viral/diagnóstico , RNA Viral/isolamento & purificação , Saliva/virologia , Técnicas de Laboratório Clínico , Humanos , Japão , Pandemias , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Manejo de Espécimes/métodos
3.
Pan Afr Med J ; 35(Suppl 2): 150, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33193965

RESUMO

The new coronavirus 2019 epidemic declared in China on December 31, 2019 soon spread to the rest of the world, becoming the subject of an unprecedented health pandemic according to the World Health Organization's declaration of March 11, 2020. It is a disease that has the potential to cause multiple systemic infections. We report here the case of an acute polyradiculoneuritis of the Guillain-Barré type (GBS) indicative of a COVID-19 infection. This is a 41 year old patient seen for ascending, symmetrical and bilateral, progressive and acute tetraparesis with in a context of influenza syndrome and digestive infections treated 2 weeks earlier. During a COVID-19 infection, certain inflammatory cells stimulated by the virus produce inflammatory cytokines creating immune-mediated processes. The same mechanism is observed in GBS being also an immune-mediated disorder. The management of this disease in COVID-19 positive patients does not differ from that of patients who do not carry the virus. The risk of respiratory distress in COVID-19 positive patients becomes twice as great in patients with GBS who test positive for COVID-19 at the same time. Monitoring for hemodynamic disorders and respiratory distress in a neuro-intensive care unit may be fruitful.


Assuntos
Betacoronavirus/patogenicidade , Infecções por Coronavirus/complicações , Síndrome de Guillain-Barré/etiologia , Pneumonia Viral/complicações , Adulto , Fibrilação Atrial/complicações , Azitromicina/uso terapêutico , Betacoronavirus/isolamento & purificação , Cloroquina/efeitos adversos , Cloroquina/uso terapêutico , Técnicas de Laboratório Clínico , Terapia Combinada , Contraindicações de Medicamentos , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/terapia , Diagnóstico Precoce , Síndrome de Guillain-Barré/diagnóstico , Síndrome de Guillain-Barré/fisiopatologia , Síndrome de Guillain-Barré/terapia , Humanos , Masculino , Debilidade Muscular/etiologia , Nasofaringe/virologia , Transtornos do Olfato/etiologia , Oxigenoterapia , Pandemias , Pneumonia Viral/diagnóstico , Pneumonia Viral/terapia , Quadriplegia/etiologia , Respiração Artificial , Incontinência Urinária/etiologia
5.
J Prev Med Hyg ; 61(3): E313-E320, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33150220

RESUMO

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is the microorganism responsible for the aggressive Coronavirus Disease (COVID-19) pandemic. During the such pandemic, discharge and community reintegration of patients are critical phases in guaranteeing public health. A review of the international and Italian experiences that represent the best available evidence was carried out, mainly focusing on the precise allocation of tasks and related responsibilities. The report provides a proposal for a systematic management pathway dedicated to COVID-19 patients. The original result is a logigramme to guide health practitioners on discharge and community reintegration of COVID-19 patients. To standardize clinical attitudes helps in ensuring quality of care and patient safety, should be a core element even during a public health emergency. The logigramme suggests, after discharge, 14 days of further isolation with regular health monitoring and, finally, the execution of a nasopharyngeal swab for identification of SARS-CoV-2 viral RNA. Home-cared patients should be placed on 7 days of further isolation after at least 2 negative RT-PCR tests for respiratory tract samples (nasopharyngeal swab). The logigramme is already used in the Department of Prevention - Local Health Agency of Lecce (Apulia) but it will be updated according to the latest research findings.


Assuntos
Infecções por Coronavirus , Monitorização Ambulatorial , Pandemias , Alta do Paciente/normas , Pneumonia Viral , Quarentena , Doenças Assintomáticas , Betacoronavirus , Técnicas de Laboratório Clínico , Convalescença , Infecções por Coronavirus/diagnóstico , Feminino , Humanos , Itália , Masculino , Nasofaringe/virologia , Fatores de Tempo
6.
Artigo em Inglês | MEDLINE | ID: mdl-33142740

RESUMO

(1) Background: The emergency linked to the spread of COVID-19 in Italy has led to inevitable consequences on the penitentiary system. The risks of this emergency in prisons is mainly related to the problem of persistent overcrowding that makes social distancing difficult and the isolation of any contagion hard to arrange. The Department of Protection for Adults and Minors of the ASL Salerno Criminal Area has taken steps in order to perform screening operations and minimize the risks for prisoners and operators. (2) Methods: We conducted a two-phase observational study. In the first phase, we offered and then executed serum COVID-19 screening to all the convicted inmates. For those who had a doubtful or positive result, a swab was executed in the shortest time possible. In the second phase, a pharyngeal swab was offered and executed to all the police officers, the penitentiary administrative staff and the medical personnel working in the prison. (3) Results: In the first phase, we executed 485 COVID-19 blood tests on prisoners, 3 (0.61%) of which were positive. The three positive inmates underwent nasopharyngeal swabbing, which ultimately were negative. After that, we executed 276 nasopharyngeal swabs on the prison personnel, penitentiary administrative staff and medical personnel-all were negative. (4) Conclusion: All tests (blood tests and swabs) that were carried out on the prisoners and on the staff were negative for COVID-19. We believe that all prisons in Italy and in the world should take action to ensure preventive and control measures in order to safeguard the health of the prison population and of all the people who work there.


Assuntos
Infecções por Coronavirus/diagnóstico , Programas de Rastreamento/estatística & dados numéricos , Nasofaringe/virologia , Pandemias , Pneumonia Viral/diagnóstico , Prisioneiros , Gestão de Riscos/métodos , Adulto , Betacoronavirus , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/prevenção & controle , Humanos , Itália , Pneumonia Viral/epidemiologia , Pneumonia Viral/prevenção & controle , Prisões
7.
Biomedica ; 40(Supl. 2): 148-158, 2020 10 30.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-33152198

RESUMO

Introduction: SARS-CoV-2 has been identified as the new coronavirus causing an outbreak of acute respiratory disease in China in December, 2019. This disease, currently named COVID-19, has been declared as a pandemic by the World Health Organization (WHO). The first case of COVID-19 in Colombia was reported on March 6, 2020. Here we characterize an early SARS-CoV-2 isolate from the pandemic recovered in April, 2020. Objective: To describe the isolation and characterization of an early SARS-CoV-2 isolate from the epidemic in Colombia. Materials and methods: A nasopharyngeal specimen from a COVID-19 positive patient was inoculated on different cell lines. To confirm the presence of SARS-CoV-2 on cultures we used qRT-PCR, indirect immunofluorescence assay, transmission and scanning electron microscopy, and next-generation sequencing. Results: We determined the isolation of SARS-CoV-2 in Vero-E6 cells by the appearance of the cytopathic effect three days post-infection and confirmed it by the positive results in the qRT-PCR and the immunofluorescence with convalescent serum. Transmission and scanning electron microscopy images obtained from infected cells showed the presence of structures compatible with SARS-CoV-2. Finally, a complete genome sequence obtained by next-generation sequencing allowed classifying the isolate as B.1.5 lineage. Conclusion: The evidence presented in this article confirms the first isolation of SARSCoV-2 in Colombia. In addition, it shows that this strain behaves in cell culture in a similar way to that reported in the literature for other isolates and that its genetic composition is consistent with the predominant variant in the world. Finally, points out the importance of viral isolation for the detection of neutralizing antibodies, for the genotypic and phenotypic characterization of the strain and for testing compounds with antiviral potential.


Assuntos
Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/virologia , Pandemias , Pneumonia Viral/virologia , RNA Viral/genética , Animais , Betacoronavirus/genética , Betacoronavirus/fisiologia , Betacoronavirus/ultraestrutura , Chlorocebus aethiops , Colômbia/epidemiologia , Convalescença , Infecções por Coronavirus/epidemiologia , Efeito Citopatogênico Viral , Técnica Indireta de Fluorescência para Anticorpo , Genoma Viral , Humanos , Microscopia Eletrônica , Tipagem Molecular , Nasofaringe/virologia , Pneumonia Viral/epidemiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de RNA , Especificidade da Espécie , Células Vero , Vírion/ultraestrutura , Cultura de Vírus
8.
Biomedica ; 40(Supl. 2): 166-172, 2020 10 30.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-33152200

RESUMO

Introduction: The 2019 coronavirus pandemic (COVID-19) has caused around 25 million cases worldwide. Asymptomatic patients have been described as potential sources of transmission. However, there are difficulties to detect them and to establish their role in the dynamics of virus transmission, which hinders the implementation of prevention strategies. Objective: To describe the behavior of asymptomatic SARS-CoV-2 virus infection in a cohort of workers at the El Dorado "Luis Carlos Galán Sarmiento" International Airport in Bogotá, Colombia. Materials and methods: A prospective cohort of 212 workers from the El Dorado airport was designed. The follow-up began in June, 2020. A survey was used to characterize health and work conditions. Every 21 day, a nasopharyngeal swab was taken to identify the presence of SARS-CoV-2 using RT-PCR. We analyzed the behavior of the cycle threshold (ORF1ab and N genes) according to the day of follow-up. Results: In the first three follow-ups of the cohort, we found an incidence of SARS-CoV-2 infection of 16.51%. The proportion of positive contacts was 14.08%. The median threshold for cycle threshold was 33.53. Conclusion: We characterized the asymptomatic SARS-CoV-2 infection in a cohort of workers. The identification of asymptomatic infected persons continues to be a challenge for epidemiological surveillance systems.


Assuntos
Aeroportos , Infecções Assintomáticas , Betacoronavirus/isolamento & purificação , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Nasofaringe/virologia , Pandemias , Pneumonia Viral/diagnóstico , Adulto , Infecções Assintomáticas/epidemiologia , Betacoronavirus/genética , Betacoronavirus/fisiologia , Colômbia , Busca de Comunicante , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas do Nucleocapsídeo/genética , Pneumonia Viral/epidemiologia , Pneumonia Viral/virologia , Estudos Prospectivos , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Inquéritos e Questionários , Proteínas Virais/genética , Replicação Viral/genética , Local de Trabalho
9.
PLoS One ; 15(11): e0242184, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33175880

RESUMO

Ivermectin has recently shown efficacy against SARS-CoV-2 in-vitro. We retrospectively reviewed severe COVID-19 patients receiving standard doses of ivermectin and we compared clinical and microbiological outcomes with a similar group of patients not receiving ivermectin. No differences were found between groups. We recommend the evaluation of high-doses of ivermectin in randomized trials against SARS-CoV-2.


Assuntos
Infecções por Coronavirus/tratamento farmacológico , Imunossupressores/uso terapêutico , Ivermectina/uso terapêutico , Pneumonia Viral/tratamento farmacológico , Adulto , Anticorpos Monoclonais Humanizados/uso terapêutico , Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/virologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Nasofaringe/virologia , Pandemias , Pneumonia Viral/diagnóstico , Pneumonia Viral/virologia , Estudos Retrospectivos , Resultado do Tratamento
10.
PLoS One ; 15(11): e0238612, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33137122

RESUMO

BACKGROUND: Rapid and extensive testing of large parts of the population and specific subgroups is crucial for proper management of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections and decision-making in times of a pandemic outbreak. However, point-of-care (POC) testing in places such as emergency units, outpatient clinics, airport security points or the entrance of any public building is a major challenge. The need for thermal cycling and nucleic acid isolation hampers the use of standard PCR-based methods for this purpose. METHODS: To avoid these obstacles, we tested PCR-independent methods for the detection of SARS-CoV-2 RNA from primary material (nasopharyngeal swabs) including reverse transcription loop-mediated isothermal amplification (RT-LAMP) and specific high-sensitivity enzymatic reporter unlocking (SHERLOCK). RESULTS: Whilst specificity of standard RT-LAMP assays appears to be satisfactory, sensitivity does not reach the current gold-standard quantitative real-time polymerase chain reaction (qPCR) assays yet. We describe a novel multiplexed RT-LAMP approach and validate its sensitivity on primary samples. This approach allows for fast and reliable identification of infected individuals. Primer optimization and multiplexing helps to increase sensitivity significantly. In addition, we directly compare and combine our novel RT-LAMP assays with SHERLOCK. CONCLUSION: In summary, this approach reveals one-step multiplexed RT-LAMP assays as a prime-option for the development of easy and cheap POC test kits.


Assuntos
Betacoronavirus/genética , Infecções por Coronavirus/diagnóstico , Técnicas de Amplificação de Ácido Nucleico/métodos , Pneumonia Viral/diagnóstico , RNA Viral/metabolismo , Betacoronavirus/isolamento & purificação , Técnicas de Laboratório Clínico , Infecções por Coronavirus/virologia , Humanos , Nasofaringe/virologia , Pandemias , Pneumonia Viral/virologia , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e Especificidade
11.
PLoS One ; 15(11): e0241740, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33137168

RESUMO

SARS-CoV-2 is spreading globally with unprecedented consequences for modern societies. The early detection of infected individuals is a pre-requisite to contain the virus. Currently, purification of RNA from patient samples followed by RT-PCR is the gold standard to assess the presence of this single-strand RNA virus. However, these procedures are time consuming, require continuous supply of specialized reagents, and are prohibitively expensive in resource-poor settings. Here, we report an improved nucleic-acid-based approach to detect SARS-CoV-2 with the ability to detect as little as five viral genome equivalents. The approach delivers results without the need to purify RNA, reduces handling steps, minimizes costs, and allows evaluation by non-specialized equipment. The use of unprocessed swap samples is enabled by employing a heat-stable RNA- and DNA-dependent DNA polymerase, which performs the double task of stringent reverse transcription of RNA at elevated temperatures as well as PCR amplification of a SARS-CoV-2 specific target gene. As results are obtained within 2 hours and can be read-out by a hand-held LED-screen, this novel protocol will be of particular importance for large-scale virus surveillance in economically constrained settings.


Assuntos
Betacoronavirus/genética , Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , RNA Viral/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/virologia , Humanos , Nasofaringe/virologia , Pandemias , Pneumonia Viral/virologia , RNA Viral/genética , Temperatura
12.
BMC Pulm Med ; 20(1): 275, 2020 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-33092563

RESUMO

BACKGROUND: Coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is an ongoing pandemic that profoundly challenges healthcare systems all over the world. Fever, cough and fatigue are the most commonly reported clinical symptoms. CASE PRESENTATION: A 58-year-old man presented at the emergency department with acute onset haemoptysis. On the fifth day after admission, he developed massive haemoptysis. Computed tomography (CT) angiography of the chest revealed alveolar haemorrhage, more prominent in the left lung. Flexible bronchoscopy confirmed bleeding from the left upper lobe, confirmed by a bronchial arteriography, which was successfully embolized. Nasopharyngeal swabs (NPS) tested for SARS-CoV-2 using real-time polymerase chain reaction (RT-PCR) repeatedly returned negative. Surprisingly, SARS-CoV-2 was eventually detected in bronchoalveolar lavage (BAL) fluid. CONCLUSIONS: Life-threatening haemoptysis is an unusual presentation of COVID-19, reflecting alveolar bleeding as a rare but possible complication. This case emphasises the added value of bronchoscopy with BAL in the diagnostic work-up in case of high clinical suspicion and negative serial NPS in patients presenting with severe symptoms.


Assuntos
Infecções por Coronavirus/complicações , Infecções por Coronavirus/diagnóstico , Hemoptise/virologia , Pneumonia Viral/complicações , Pneumonia Viral/diagnóstico , Doença Aguda , Betacoronavirus , Líquido da Lavagem Broncoalveolar/virologia , Broncoscopia , Angiografia por Tomografia Computadorizada , Hemoptise/diagnóstico por imagem , Humanos , Masculino , Pessoa de Meia-Idade , Nasofaringe/virologia , Pandemias
13.
Ann Ital Chir ; 91: 345-351, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33055390

RESUMO

INTRODUCTION: The recent Sars-CoV2 pandemic has dramatically slowed patients' access to our clinic for vascular pathology when the contagion curve peaked. The need to restore the assistance activity has led us to adopt new individual prophylaxis and hygiene measures. METHODS: Doctors and staff must wear dedicated clothes. Mask and gloves are mandatory for patients. A visit is scheduled every 60 minutes to allow the sanitation of the rooms. The day before the visit patients are contacted by telephone for the Covid-19 risk triage. In the presence of symptoms the visit is postponed. In the presence of other risk factors a IgG/IgM Rapid Test for Covid-19 is performed on admission to the clinic. In the presence of fever, if an extraordinary rapid test cannot be performed, the visit must be postponed. Rapid test positive patients cannot be visited: they are placed in solitary confinement at their home waiting for a nasopharyngeal swab for Covid-19. When the rapid test is positive, immediate room sanitation also occurs. The rooms dedicated to the outpatient clinic as well as medical and not medical instruments are disinfected. CONCLUSION: The one adopted can be a useful management model for any type of care activity in order to guarantee the safety of patients and all the staff. KEY WORDS: COVID-19, Management, vascular, Outpatient clinic.


Assuntos
Betacoronavirus , Cardiologia/organização & administração , Técnicas de Laboratório Clínico , Infecções por Coronavirus/prevenção & controle , Controle de Infecções/métodos , Ambulatório Hospitalar/organização & administração , Pandemias/prevenção & controle , Pneumonia Viral/prevenção & controle , Anticorpos Antivirais/sangue , Agendamento de Consultas , Betacoronavirus/imunologia , Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/epidemiologia , Desinfecção , Formulários como Assunto , Hospitais Universitários/organização & administração , Humanos , Higiene/normas , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Controle de Infecções/organização & administração , Controle de Infecções/normas , Itália/epidemiologia , Nasofaringe/virologia , Equipamento de Proteção Individual , Pneumonia Viral/diagnóstico , Pneumonia Viral/epidemiologia , Guias de Prática Clínica como Assunto , Avaliação de Sintomas , Termometria , Triagem/organização & administração
14.
PLoS Biol ; 18(10): e3000896, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-33006983

RESUMO

The ongoing COVID-19 pandemic has created an unprecedented need for rapid diagnostic testing. The World Health Organization (WHO) recommends a standard assay that includes an RNA extraction step from a nasopharyngeal (NP) swab followed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) to detect the purified SARS-CoV-2 RNA. The current global shortage of RNA extraction kits has caused a severe bottleneck to COVID-19 testing. The goal of this study was to determine whether SARS-CoV-2 RNA could be detected from NP samples via a direct RT-qPCR assay that omits the RNA extraction step altogether. The direct RT-qPCR approach correctly identified 92% of a reference set of blinded NP samples (n = 155) demonstrated to be positive for SARS-CoV-2 RNA by traditional clinical diagnostic RT-qPCR that included an RNA extraction. Importantly, the direct method had sufficient sensitivity to reliably detect those patients with viral loads that correlate with the presence of infectious virus. Thus, this strategy has the potential to ease supply choke points to substantially expand COVID-19 testing and screening capacity and should be applicable throughout the world.


Assuntos
Betacoronavirus/genética , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , RNA Viral/genética , Kit de Reagentes para Diagnóstico/normas , Reação em Cadeia da Polimerase Via Transcriptase Reversa/normas , Betacoronavirus/patogenicidade , Técnicas de Laboratório Clínico/normas , Infecções por Coronavirus/virologia , Primers do DNA/normas , Humanos , Nasofaringe/virologia , Pandemias , Pneumonia Viral/virologia , Sensibilidade e Especificidade , Estados Unidos , Carga Viral
15.
PLoS Biol ; 18(10): e3000867, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-33027248

RESUMO

The current quantitative reverse transcription PCR (RT-qPCR) assay recommended for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) testing in the United States requires analysis of 3 genomic targets per sample: 2 viral and 1 host. To simplify testing and reduce the volume of required reagents, we devised a multiplex RT-qPCR assay to detect SARS-CoV-2 in a single reaction. We used existing N1, N2, and RP primer and probe sets by the Centers for Disease Control and Prevention, but substituted fluorophores to allow multiplexing of the assay. The cycle threshold (Ct) values of our multiplex RT-qPCR were comparable to those obtained by the single assay adapted for research purposes. Low copy numbers (≥500 copies/reaction) of SARS-CoV-2 RNA were consistently detected by the multiplex RT-qPCR. Our novel multiplex RT-qPCR improves upon current single diagnostics by saving reagents, costs, time, and labor.


Assuntos
Betacoronavirus/genética , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/diagnóstico , Reação em Cadeia da Polimerase Multiplex/normas , Pneumonia Viral/diagnóstico , RNA Viral/genética , Kit de Reagentes para Diagnóstico/normas , Reação em Cadeia da Polimerase Via Transcriptase Reversa/normas , Betacoronavirus/patogenicidade , Estudos de Casos e Controles , Técnicas de Laboratório Clínico/normas , Infecções por Coronavirus/virologia , Primers do DNA/normas , Células HEK293 , Humanos , Limite de Detecção , Nasofaringe/virologia , Pandemias , Pneumonia Viral/virologia , Estados Unidos
16.
Aust J Gen Pract ; 49(11): 728-732, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-33123703

RESUMO

BACKGROUND: General practitioners (GPs) have some of the highest rates of mortality from COVID-19 among healthcare workers. SARS-CoV-2 has unique properties that place GPs at particular risk. OBJECTIVE: The aim of this article is to discuss the nose-related features of SARS-CoV-2 that place GPs at risk, and to make recommendations pertinent to the safety and protection of primary healthcare physicians. DISCUSSION: The highest viral load of SARS-CoV-2 is in the nose and nasopharynx. It is often highest early in the illness, before the development of symptoms. Further, SARS-CoV-2 replicates and continues to shed in the nasopharynx long after the virus is no longer detectable in the lower respiratory tract. This places any physician performing examinations on, or procedures involving, the upper respiratory tract at risk for contracting COVID-19. New-onset hyposmia and dysgeusia are indicators for COVID-19 and should be included in screening protocols.


Assuntos
Betacoronavirus , Infecções por Coronavirus , Clínicos Gerais/estatística & dados numéricos , Controle de Infecções , Cavidade Nasal/virologia , Nasofaringe/virologia , Transtornos do Olfato/virologia , Pandemias , Pneumonia Viral , Austrália , Betacoronavirus/isolamento & purificação , Betacoronavirus/fisiologia , Infecções por Coronavirus/mortalidade , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/transmissão , Infecções por Coronavirus/virologia , Transmissão de Doença Infecciosa/prevenção & controle , Humanos , Controle de Infecções/instrumentação , Controle de Infecções/métodos , Controle de Infecções/normas , Mucosa Nasal/metabolismo , Exposição Ocupacional/prevenção & controle , Exposição Ocupacional/estatística & dados numéricos , Pandemias/prevenção & controle , Equipamento de Proteção Individual/provisão & distribução , Pneumonia Viral/mortalidade , Pneumonia Viral/prevenção & controle , Pneumonia Viral/transmissão , Pneumonia Viral/virologia , Guias de Prática Clínica como Assunto , Atenção Primária à Saúde/normas , Gestão de Riscos , Carga Viral
17.
Euro Surveill ; 25(42)2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-33094715

RESUMO

An outbreak of 59 cases of coronavirus disease (COVID-19) originated with 13 cases linked by a 7 h, 17% occupancy flight into Ireland, summer 2020. The flight-associated attack rate was 9.8-17.8%. Spread to 46 non-flight cases occurred country-wide. Asymptomatic/pre-symptomatic transmission in-flight from a point source is implicated by 99% homology across the virus genome in five cases travelling from three different continents. Restriction of movement on arrival and robust contact tracing can limit propagation post-flight.


Assuntos
Viagem Aérea , Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/epidemiologia , Pandemias , Pneumonia Viral/epidemiologia , Doença Relacionada a Viagens , Doenças Assintomáticas , Betacoronavirus/genética , Busca de Comunicante , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/transmissão , Infecções por Coronavirus/virologia , Surtos de Doenças , Exposição Ambiental , Características da Família , Controle de Formulários e Registros , Genoma Viral , Hospitalização , Humanos , Controle de Infecções/métodos , Irlanda/epidemiologia , Nasofaringe/virologia , Pandemias/prevenção & controle , Pneumonia Viral/prevenção & controle , Pneumonia Viral/transmissão , Pneumonia Viral/virologia , RNA Viral/genética , Estações do Ano , Homologia de Sequência do Ácido Nucleico , Fatores de Tempo , Sequenciamento Completo do Genoma
20.
G Ital Nefrol ; 37(5)2020 Oct 05.
Artigo em Italiano | MEDLINE | ID: mdl-33026198

RESUMO

We report the brief experience of the Nephrology Center located in a "no-COVID" Hospital in Massa Marittima. We describe the actions taken to prevent the transmission of the virus SARS-CoV-2 among hemodialysis patients and healthcare workers and the methods for diagnosing COVID-19, with particular attention to serological tests and nasopharyngeal swabs in asymptomatic subjects. The detection of IgM and IgG antibodies through the serological test performed on 34 patients, all negative for nasopharyngeal swabs, showed positivity in 41,18% of cases. These have been classified as false positives following repeated negative nasopharyngeal swabs, the evaluation of clinical and epidemiological history and of clinical manifestations and, finally, a second serological test performed after 18 days, which resulted negative for all patients. Interpreting serological tests is not easy; the strategies for diagnosis should include clinical and epidemiological history and clinical manifestations, as well as the results of confirmation tests and the evaluation over a precise observation period. Otherwise, there is a risk of considering as protected by antibodies subjects that are in fact false positives.


Assuntos
Anticorpos Antivirais/sangue , Betacoronavirus/imunologia , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Unidades Hospitalares de Hemodiálise/estatística & dados numéricos , Pandemias , Pneumonia Viral/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Doenças Assintomáticas , Betacoronavirus/isolamento & purificação , Reações Falso-Positivas , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imunoglobulina M/sangue , Imunoglobulina M/imunologia , Itália , Masculino , Pessoa de Meia-Idade , Nasofaringe/virologia , Guias de Prática Clínica como Assunto , Reprodutibilidade dos Testes , Estudos Retrospectivos , Sensibilidade e Especificidade
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