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2.
Euro Surveill ; 25(3)2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31992388

RESUMO

A novel coronavirus (2019-nCoV) causing severe acute respiratory disease emerged recently in Wuhan, China. Information on reported cases strongly indicates human-to-human spread, and the most recent information is increasingly indicative of sustained human-to-human transmission. While the overall severity profile among cases may change as more mild cases are identified, we estimate a risk of fatality among hospitalised cases at 14% (95% confidence interval: 3.9-32%).


Assuntos
Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/transmissão , Coronavirus/isolamento & purificação , Surtos de Doenças , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/transmissão , China/epidemiologia , Doenças Transmissíveis Emergentes/epidemiologia , Doenças Transmissíveis Emergentes/transmissão , Coronavirus/classificação , Infecções por Coronavirus/mortalidade , Mortalidade Hospitalar , Hospitalização , Humanos , Saúde Pública , Medição de Risco
3.
Euro Surveill ; 25(3)2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31992387

RESUMO

BackgroundThe ongoing outbreak of the recently emerged novel coronavirus (2019-nCoV) poses a challenge for public health laboratories as virus isolates are unavailable while there is growing evidence that the outbreak is more widespread than initially thought, and international spread through travellers does already occur.AimWe aimed to develop and deploy robust diagnostic methodology for use in public health laboratory settings without having virus material available.MethodsHere we present a validated diagnostic workflow for 2019-nCoV, its design relying on close genetic relatedness of 2019-nCoV with SARS coronavirus, making use of synthetic nucleic acid technology.ResultsThe workflow reliably detects 2019-nCoV, and further discriminates 2019-nCoV from SARS-CoV. Through coordination between academic and public laboratories, we confirmed assay exclusivity based on 297 original clinical specimens containing a full spectrum of human respiratory viruses. Control material is made available through European Virus Archive - Global (EVAg), a European Union infrastructure project.ConclusionThe present study demonstrates the enormous response capacity achieved through coordination of academic and public laboratories in national and European research networks.


Assuntos
Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/virologia , Coronavirus/classificação , Coronavirus/genética , Técnicas de Laboratório Clínico/métodos , Coronavirus/isolamento & purificação , Surtos de Doenças , Humanos , RNA Viral/análise , Reação em Cadeia da Polimerase em Tempo Real/métodos , Sensibilidade e Especificidade
4.
Emerg Microbes Infect ; 8(1): 1528-1534, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31645223

RESUMO

Dromedary camels are important reservoir hosts of various coronaviruses, including Middle East respiratory syndrome coronavirus (MERS-CoV) that cause human infections. CoV genomes regularly undergo recombination during infection as observed in bat SARS-related CoVs. Here we report for the first time that only a small proportion of MERS-CoV receptor-binding domain positive (RBD) of spike protein positive camel sera in Kenya were also seropositive to MERS-CoV nucleocapsid (NP). In contrast, many of them contain antibodies against bat HKU8-related (HKU8r)-CoVs. Among 584 camel samples that were positive against MERS-CoV RBD, we found only 0.48 (8.22%) samples were also positive for NP. Furthermore, we found bat HKU8r-CoV NP antibody in 73 (12.5%) of the MERS-CoV RBD positive and NP negative samples, yet found only 3 (0.43%) of the HKU8r-CoV S1 antibody in the same samples. These findings may indicate co-infection with MERS-CoV and a HKU8r-CoV in camels. It may also raise the possibility of the circulation of a recombinant coronavirus virus with the spike of MERS-CoV and the NP of a HKU8r-CoV in Kenya. We failed to find molecular evidence of an HKU8r-CoV or a putative recombinant virus. Our findings should alert other investigators to look for molecular evidence of HKU8r-CoV or recombinants.


Assuntos
Camelus/virologia , Infecções por Coronavirus/veterinária , Coronavirus/imunologia , Coronavírus da Síndrome Respiratória do Oriente Médio/imunologia , Animais , Anticorpos Antivirais/sangue , Camelus/sangue , Quirópteros/virologia , Coronavirus/genética , Coronavirus/isolamento & purificação , Infecções por Coronavirus/sangue , Infecções por Coronavirus/virologia , Quênia , Coronavírus da Síndrome Respiratória do Oriente Médio/genética , Coronavírus da Síndrome Respiratória do Oriente Médio/isolamento & purificação , Proteínas do Nucleocapsídeo/imunologia , Recombinação Genética , Glicoproteína da Espícula de Coronavírus/imunologia
5.
Aust Vet J ; 97(10): 418-421, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31556108

RESUMO

This study reports the prevalence of potential faecal pathogens in the microbiome detected in a cohort of cats and dogs with diarrhoea in Perth, Western Australia. Records from a commercial diagnostic laboratory using faecal PCR testing between July 2014 and August 2015 were reviewed.Of 289 feline faecal samples reviewed, Salmonella spp. (1.7%), Campylobacter spp. (47.6%), Clostridium perfringens (81.3%), Giardia spp. (11.1%), Toxoplasma gondii (1.2%), Tritrichomonas foetus (4.8%), panleukopenia virus (6.5%) and coronavirus (39.5%) were detected. In dogs, Salmonella spp. (5.4%), Campylobacter spp. (36.3%), C. perfringens (85.4%), Giardia spp. (6.2%), parvovirus (9.4%), coronavirus (4.7%) and distemper virus (1.5%) were detected.


Assuntos
Doenças do Gato/microbiologia , Diarreia/veterinária , Doenças do Cão/microbiologia , Microbioma Gastrointestinal , Animais , Doenças do Gato/epidemiologia , Gatos , Coronavirus/isolamento & purificação , Diarreia/epidemiologia , Diarreia/microbiologia , Doenças do Cão/epidemiologia , Cães , Fezes/microbiologia , Feminino , Giardia/isolamento & purificação , Bactérias Gram-Negativas/isolamento & purificação , Masculino , Reação em Cadeia da Polimerase/veterinária , Austrália Ocidental/epidemiologia
6.
Med. clín (Ed. impr.) ; 153(5): 205-212, sept. 2019. tab
Artigo em Espanhol | IBECS | ID: ibc-183997

RESUMO

El incremento de los viajes internacionales, la creciente presencia de vectores transmisores de arbovirus en nuestro país, las alertas de fiebres hemorrágicas, como el actual brote de ébola en la R. D. del Congo y los casos autóctonos de fiebre hemorrágica de Crimea-Congo en nuestro país, ponen de nuevo en primer plano las enfermedades tropicales. El aislamiento de los casos sospechosos de enfermedades de alta transmisibilidad y letalidad ha de ser una prioridad (fiebres hemorrágicas, MERS-CoV). Al valorar al paciente, una cuidadosa historia clínica basada en los aspectos epidemiológicos de la zona de procedencia, las actividades realizadas, el tiempo de estancia en el mismo y el inicio de los síntomas nos ayudarán finalmente, si no al diagnóstico definitivo, sí al menos a descartar las enfermedades que signifiquen una amenaza para él. Por su frecuencia y gravedad la malaria debe ser descartada, sin olvidar las otras causas habituales de fiebre con las que el médico de urgencias debe estar familiarizado también


The increase in international travel, the growing presence of arbovirus vectors in our country, and notifications of haemorrhagic fever such as the current outbreak of Ebola in D.R. Congo and the cases of Crimea-Congo haemorrhagic fever in our country have again cast the spotlight on tropical diseases Isolating suspected cases of highly contagious and lethal diseases must be a priority (Haemorrhagic fever, MERS-CoV). Assessing the patient, taking a careful medical history based on epidemiological aspects of the area of origin, activities they have carried out, their length of stay in the area and the onset of symptoms, will eventually help us, if not to make a definitive diagnosis, at least to exclude diseases that pose a threat to these patients. Malaria should be ruled out because of its frequency, without forgetting other common causes of fever familiar to emergency doctors


Assuntos
Humanos , Controle Sanitário de Viajantes , Doença Relacionada a Viagens , Isolamento de Pacientes/tendências , Saúde do Viajante , Doenças Transmissíveis Importadas/epidemiologia , Febres Hemorrágicas Virais/epidemiologia , Infecções por Arbovirus/epidemiologia , Arbovirus/isolamento & purificação , Meningite/epidemiologia , Febre Tifoide/epidemiologia , Rickettsia/isolamento & purificação , Coronavirus/isolamento & purificação , Esquistossomose/epidemiologia
7.
Tuberk Toraks ; 67(2): 124-130, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31414643

RESUMO

Introduction: Exacerbations of chronic obstructive pulmonary disease (COPD) are often caused by respiratory tract infections. The aim of this study was to investigate the clinical, laboratory and computed tomography features of patients with hospitalized COPD exacerbations in which respiratory viruses were detected using a real-time polymerase chain reaction (PCR) technique. Materials and Methods: This retrospectively planned study included patients hospitalized in the chest diseases clinic due to exacerbation of COPD between November 2018-February 2019. The study included patients who had virus-specific real-time PCR, and computed tomography scans of the chest. Result: A total of 110 patients were included in the study. Respiratory viruses were identified in the nasopharyngeal swabs of 50 patients (45.5%) using the real-time PCR method, with rhinovirus (25%), influenza A (13.1%) and coronavirus (11.8%) being the most commonly isolated agents. The mean age of the patients was 68.28 ± 9.59 years in the virus-positive group and 68.20 ± 8.27 years in the virus-negative group (p= 0.963). Gender distribution, rate of smokers, exposure to biofuels, blood leukocyte count, neutrophil percentage, C-reactive protein (CRP) level, FEV1/FVC ratio did not significantly differ between the two groups (p> 0.05). Procalcitonin (PCT) and FEV1 values were significantly lower (p= 0.001 and p= 0.028, respectively) and the number of exacerbations was significantly higher in the virus-positive group (p= 0.001). The length of hospital stay was longer in the virus-positive group than in the virus-negative group (p= 0.012). Among the findings of computed tomography (CT) of the chest, bronchial wall thickening, cystic bronchiectasis, and emphysema did not differ significantly (p> 0.05). The rate of infiltrative lesions (tree-in-bud opacity, ground-glass opacity, atypical pneumonia) was significantly higher in the virus-positive group (p= 0.020). Conclusions: Viral respiratory tract infections should be considered in hospitalized patients with an exacerbation of COPD who have a history of frequent exacerbations, normal PCT value, and the absence of consolidation in CT scan of the chest. The use of broadspectrum antibiotic therapy should be avoided in patients with these features.


Assuntos
Doença Pulmonar Obstrutiva Crônica/complicações , Infecções Respiratórias/complicações , Viroses/complicações , Idoso , Bronquiectasia , Coronavirus/isolamento & purificação , Feminino , Humanos , Vírus da Influenza A/isolamento & purificação , Influenza Humana/complicações , Tempo de Internação , Masculino , Pessoa de Meia-Idade , Nasofaringe/virologia , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Doença Pulmonar Obstrutiva Crônica/virologia , Reação em Cadeia da Polimerase em Tempo Real , Testes de Função Respiratória , Infecções Respiratórias/virologia , Estudos Retrospectivos , Rhinovirus/isolamento & purificação , Tomografia Computadorizada por Raios X , Viroses/virologia
8.
Virus Genes ; 55(4): 520-531, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31129785

RESUMO

Porcine deltacoronavirus (PDCoV) is an emerging swine enteropathogenic coronavirus that causes watery diarrhea, vomiting and mortality in newborn piglets. Previous studies have suggested that PDCoV infection antagonizes RIG-I-like receptor (RLR)-mediated IFN-ß production to evade host innate immune defense, and PDCoV-encoded nonstructural protein nsp5 and accessory protein NS6 are associated with this process. However, whether the structural protein(s) of PDCoV also antagonize IFN-ß production remains unclear. In this study, we found that PDCoV nucleocapsid (N) protein, the most abundant viral structural protein, suppressed Sendai virus (SEV)-induced IFN-ß production and transcription factor IRF3 activation, but did not block IFN-ß production induced by overexpressing RIG-I/MDA5. Furthermore, study revealed that PDCoV N protein interacted with RIG-I and MDA5 in an in vitro overexpression system and evident interactions between N protein and RIG-I could be detected in the context of PDCoV infection, which interfered with the binding of dsRNA and protein activator of protein kinase R (PACT) to RIG-I. Together, our results demonstrate that PDCoV N protein is an IFN antagonist and utilizes diverse strategies to attenuate RIG-I recognition and activation.


Assuntos
Coronavirus/imunologia , Proteína DEAD-box 58/antagonistas & inibidores , Interferon beta/antagonistas & inibidores , Proteínas do Nucleocapsídeo/imunologia , Suínos/virologia , Animais , Coronavirus/genética , Coronavirus/isolamento & purificação , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/virologia , Células HEK293 , Humanos , Fator Regulador 3 de Interferon/antagonistas & inibidores , Interferon beta/genética , Ligação Proteica , RNA de Cadeia Dupla/antagonistas & inibidores , Proteínas de Ligação a RNA/antagonistas & inibidores , Vírus Sendai/imunologia , Doenças dos Suínos/virologia
9.
Appl Microbiol Biotechnol ; 103(12): 4943-4952, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31025076

RESUMO

Swine enteric coronaviruses are a group of most significant pathogens causing diarrhea in piglets with similar clinical symptoms and pathological changes. To develop a simple, rapid, accurate, and high-throughput detection method for diagnosis and differential diagnosis on swine enteric coronaviruses, specific primers and probes were designed based on the highly conserved regions of transmissible gastroenteritis virus (TGEV) N, porcine epidemic diarrhea virus (PEDV) M, porcine deltacoronavirus (PDCoV) M, and porcine enteric alphacoronavirus (PEAV) N genes respectively. A TaqMan-probe-based multiplex real-time RT-qPCR assay was developed and optimized to simultaneously detect these swine enteric coronaviruses. The results showed that the limit of detection can reach as low as 10 copies in singular real-time RT-qPCR assays and 100 copies in multiplex real-time RT-qPCR assay, with all correlation coefficients (R2) at above 0.99, and the amplification efficiency at between 90 and 120%. This multiplex real-time RT-qPCR assay demonstrated high sensitivity, extreme specificity, and excellent repeatability. The multiplex real-time RT-qPCR assay was then employed to detect the swine enteric coronavirus from 354 field diarrheal samples. The results manifested that TGEV and PDCoV were the main pathogens in these samples, accompanied by co-infections. This well-established multiplex real-time RT-qPCR assay provided a rapid, efficient, specific, and sensitive tool for detection of swine enteric coronaviruses.


Assuntos
Infecções por Coronavirus/veterinária , Coronavirus/isolamento & purificação , Reação em Cadeia da Polimerase Multiplex/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Doenças dos Suínos/diagnóstico , Animais , Coronavirus/classificação , Infecções por Coronavirus/diagnóstico , Primers do DNA/genética , Diagnóstico Diferencial , Diarreia/virologia , Fezes/virologia , Gastroenterite Suína Transmissível/diagnóstico , Gastroenterite Suína Transmissível/virologia , Limite de Detecção , Vírus da Diarreia Epidêmica Suína/genética , Vírus da Diarreia Epidêmica Suína/isolamento & purificação , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Suínos , Doenças dos Suínos/virologia , Vírus da Gastroenterite Transmissível/genética , Vírus da Gastroenterite Transmissível/isolamento & purificação
10.
Braz J Microbiol ; 50(2): 547-556, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30877663

RESUMO

Coronaviruses are single-stranded positive-sense RNA viruses associated with important avian diseases. Their relatively high rates of mutation and recombination frequencies allow them to adapt to new hosts and ecological niches. Although Brazil has 18% of global avian species diversity, studies regarding the presence of avian viral diseases in wild birds in South America are scarce. In this study, we performed a retrospective analysis of the presence of CoVs in 746 wild birds. Oropharyngeal and cloacal swabs were obtained and placed together in vials containing VTM transport medium collected in different regions of Brazil between 2006 and 2013. Screening for viral nucleic acid was performed using conventional RT-PCR and pancoronavirus nested PCR. Positive samples were characterized by partial sequencing of the RNA-dependent RNA polymerase (RdRp) gene, and ensuing phylogenetic analysis was performed to investigate the association between virus epidemiology and bird migration routes. Coronavirus RNA were detected and sequenced from six samples, in which three were related to gammacoronaviruses group and the other three to deltacoronavirus group. Our study documents the presence of CoVs related to avian gamma- and deltacoronaviruses circulating in both urban- and poultry-farm regions of Brazil, implicating wild birds as potential carriers of CoVs which may represent a risk to poultry farms and public health in Brazil.


Assuntos
Doenças das Aves/epidemiologia , Aves/virologia , Infecções por Coronavirus/epidemiologia , Coronavirus/isolamento & purificação , Reservatórios de Doenças/virologia , Gammacoronavirus/isolamento & purificação , Animais , Animais Selvagens/virologia , Sequência de Bases , Doenças das Aves/virologia , Brasil/epidemiologia , Coronavirus/genética , Gammacoronavirus/genética , Filogenia , RNA Replicase/genética , RNA Viral/genética , Estudos Retrospectivos
11.
Viruses ; 11(3)2019 03 02.
Artigo em Inglês | MEDLINE | ID: mdl-30832341

RESUMO

During the past two decades, three zoonotic coronaviruses have been identified as the cause of large-scale disease outbreaks⁻Severe Acute Respiratory Syndrome (SARS), Middle East Respiratory Syndrome (MERS), and Swine Acute Diarrhea Syndrome (SADS). SARS and MERS emerged in 2003 and 2012, respectively, and caused a worldwide pandemic that claimed thousands of human lives, while SADS struck the swine industry in 2017. They have common characteristics, such as they are all highly pathogenic to humans or livestock, their agents originated from bats, and two of them originated in China. Thus, it is highly likely that future SARS- or MERS-like coronavirus outbreaks will originate from bats, and there is an increased probability that this will occur in China. Therefore, the investigation of bat coronaviruses becomes an urgent issue for the detection of early warning signs, which in turn minimizes the impact of such future outbreaks in China. The purpose of the review is to summarize the current knowledge on viral diversity, reservoir hosts, and the geographical distributions of bat coronaviruses in China, and eventually we aim to predict virus hotspots and their cross-species transmission potential.


Assuntos
Quirópteros/virologia , Coronavirus/genética , Reservatórios de Doenças/virologia , Evolução Molecular , Zoonoses/virologia , Animais , China/epidemiologia , Coronavirus/isolamento & purificação , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/veterinária , Surtos de Doenças/prevenção & controle , Variação Genética , Genoma Viral , Humanos , Filogenia , Zoonoses/epidemiologia , Zoonoses/transmissão
12.
Viruses ; 11(2)2019 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-30791586

RESUMO

Bats are a unique group of mammals of the order Chiroptera. They are highly diversified and are the group of mammals with the second largest number of species. Such highly diversified cell types and receptors facilitate them to be potential hosts of a large variety of viruses. Bats are the only group of mammals capable of sustained flight, which enables them to disseminate the viruses they harbor and enhance the chance of interspecies transmission. This article aims at reviewing the various aspects of the global epidemiology of bat coronaviruses (CoVs). Before the SARS epidemic, bats were not known to be hosts for CoVs. In the last 15 years, bats have been found to be hosts of >30 CoVs with complete genomes sequenced, and many more if those without genome sequences are included. Among the four CoV genera, only alphaCoVs and betaCoVs have been found in bats. As a whole, both alphaCoVs and betaCoVs have been detected from bats in Asia, Europe, Africa, North and South America and Australasia; but alphaCoVs seem to be more widespread than betaCoVs, and their detection rate is also higher. For betaCoVs, only those from subgenera Sarbecovirus, Merbecovirus, Nobecovirus and Hibecovirus have been detected in bats. Most notably, horseshoe bats are the reservoir of SARS-CoV, and several betaCoVs from subgenus Merbecovirus are closely related to MERS-CoV. In addition to the interactions among various bat species themselves, bat⁻animal and bat⁻human interactions, such as the presence of live bats in wildlife wet markets and restaurants in Southern China, are important for interspecies transmission of CoVs and may lead to devastating global outbreaks.


Assuntos
Quirópteros/virologia , Infecções por Coronavirus/veterinária , Coronavirus/genética , Reservatórios de Doenças/virologia , Evolução Molecular , África/epidemiologia , Animais , Ásia/epidemiologia , Australásia/epidemiologia , China/epidemiologia , Coronavirus/isolamento & purificação , Infecções por Coronavirus/epidemiologia , Europa (Continente)/epidemiologia , Genoma Viral , Saúde Global , Humanos , América do Norte/epidemiologia , Filogenia , Filogeografia , América do Sul/epidemiologia
13.
Transbound Emerg Dis ; 66(3): 1324-1331, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30801935

RESUMO

Porcine deltacoronavirus (PDCoV) has emerged and spread throughout the porcine industry in many countries over the last 6 years. PDCoV caused watery diarrhoea, vomiting and dehydration in newborn piglets. A sensitive diagnostic method would be beneficial to the prevention and control of PDCoV infection. Recombinase polymerase amplification (RPA) is an isothermal amplification method which has been widely used for virus detection. A probe-based reverse transcription RPA (RT-RPA) assay was developed for real-time detection of PDCoV. The amplification can be finished in 20 min and fluorescence monitoring was performed by a portable device. The lowest detection limit of the PDCoV RT-RPA assay was 100 copies of RNA molecules per reaction; moreover, the RT-RPA assay had no cross-reaction with other common swine viruses. The clinical performance of the RT-RPA assay was evaluated using 108 clinical samples (54 intestine specimens and 54 faecal swab specimens). The coincidence rate of the detection results for clinical samples between RT-RPA and RT-qPCR was 97.2%. In summary, the real-time RT-RPA assay offers a promising alternative to RT-qPCR for point-of-care detection of PDCoV.


Assuntos
Infecções por Coronavirus/veterinária , Coronavirus/isolamento & purificação , Sistemas Automatizados de Assistência Junto ao Leito , Doenças dos Suínos/diagnóstico , Animais , Coronavirus/genética , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/virologia , Limite de Detecção , Recombinases/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Sensibilidade e Especificidade , Suínos , Doenças dos Suínos/virologia , Fatores de Tempo
14.
Virus Res ; 260: 60-66, 2019 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-30447246

RESUMO

Coronaviruses (CoVs) have been detected worldwide in several bat species, which are considered the main reservoir. The attention to the high diversity of CoVs hosted by bats has increased during the last decade due to the high number of human infections caused by two zoonotic Beta-CoVs, SARS-CoV and MERS-CoV, that cause several respiratory diseases. Among coronaviruses, two Alpha-CoV strains (HuCoV-229E and HuCoV-NL63) cause mild respiratory disease that can change to severe disease in children, elderly and individuals affected by illnesses. Phylogenetic analysis conducted on bat Alpha-CoV strains revealed their evolutive correlation to human strains, suggesting their origin in bats. The genome of CoVs is characterized by a high frequency of mutations and recombination events, increasing their ability to switch hosts and their zoonotic potential. In this study, three strains of Alpha-CoV genera detected in Italian bats (Pipistrellus kuhlii) were fully sequenced by Next Generation Sequencing (NGS) and characterized. The complete genome analysis showed the correlation of the Italians strains with a Chinese strain detected in 2013 and, based on CoV molecular species demarcation, two new Alpha-CoV species were established. The analysis of a fragment of the RNA-dependent RNA polymerase (RdRp) showed the correlation of the Italian strains with CoVs that was only detected in the bat Pipistrellus genera (Pipistrellus kuhlii and Pipistrellus Pipistrellus) in European countries.


Assuntos
Quirópteros/virologia , Infecções por Coronavirus/veterinária , Coronavirus/genética , Coronavirus/isolamento & purificação , Genoma Viral , Animais , Biologia Computacional , Coronavirus/classificação , Infecções por Coronavirus/virologia , Sequenciamento de Nucleotídeos em Larga Escala , Itália , Filogenia , Homologia de Sequência
15.
Arch Virol ; 164(2): 413-425, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30377826

RESUMO

Porcine deltacoronavirus (PDCoV) was first detected in Hong Kong and has recently spread to many countries around the world. PDCoV causes acute diarrhea and vomiting in pigs, resulting in significant economic losses in the global pork industry. In this study, a Chinese PDCoV strain, designated CHN-HG-2017, was isolated from feces of a suckling piglet with severe watery diarrhea on a farm located in central China. Subsequently, the virus was identified by an indirect immunofluorescence assay and electron microscopy. A nucleotide sequence alignment showed that the whole genome of CHN-HG-2017 is 97.6%-99.1% identical to other PDCoV strains. Analysis of potential recombination sites showed that CHN-HG-2017 is a possible recombinant originating from the strains CH/SXD1/2015 and Vietnam/HaNoi6/2015. Furthermore, the pathogenicity of this recombinant PDCoV strain was investigated in 5-day-old piglets by oral inoculation. The challenged piglets developed typical symptoms, such as vomiting, anorexia, diarrhea and lethargy, from 1 to 7 days post-inoculation (DPI). Viral shedding was detected in rectal swabs until 14 DPI in the challenged piglets. Interestingly, high titers of virus-neutralizing antibodies in sera were detected at 21 DPI. Tissues of small intestines from CHN-HG-2017-infected piglets at 4 DPI displayed significant macroscopic and microscopic lesions with clear viral antigen expression. Our analysis of the full genome sequence of a recombinant PDCoV and its virulence in suckling piglets might provide new insights into the pathogenesis of PDCoV and facilitate further investigation of this newly emerged pathogen.


Assuntos
Infecções por Coronavirus/veterinária , Coronavirus/isolamento & purificação , Coronavirus/patogenicidade , Doenças dos Suínos/virologia , Animais , China , Coronavirus/classificação , Coronavirus/genética , Infecções por Coronavirus/virologia , Diarreia/veterinária , Diarreia/virologia , Fezes/virologia , Genoma Viral , Genômica , Filogenia , Suínos , Vietnã , Virulência
16.
Viral Immunol ; 32(1): 63-67, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30207926

RESUMO

This case study report describes a transmissible gastroenteritis coronavirus (TGEV) infection presented in a commercial pig herd. The clinical signs of infection appeared in newborn piglets, including medium morbidity and low mortality rates. Rectal swabs were collected from five different affected litters for laboratory examinations. Samples from two dead piglets and two euthanized affected piglets were collected for gross and histopathological examinations. All fecal samples were tested TGEV positive by real-time polymerase chain reaction (RT-PCR). Necropsy revealed nonspecific gross lesions. The histopathological examinations revealed villi fused with denuded tips and severe villus atrophy, leading to extensive epithelial flattening in middle and lower small intestine. The architecture pattern of villi presented columnar and cuboidal poorly differentiated enterocytes with mild subepithelial edema. In some enterocytes, pycnotic nuclei were detected. Microscopic examination of brain tissue revealed diffuse gliosis in the area of pia matter with mild congestion of the meningeal and parenchymal vessels and neuronal degeneration. In conclusion, this case study reported an epidemic TGEV infection in piglets, characterized by low mortality and medium morbidity rates accompanied by typical histopathological lesions in small intestine, as well as by coexisting brain lesions, that are described for the first time.


Assuntos
Encéfalo/patologia , Encéfalo/virologia , Diarreia/veterinária , Gastroenterite Suína Transmissível/diagnóstico , Intestino Delgado/virologia , Animais , Coronavirus/genética , Coronavirus/isolamento & purificação , Diarreia/virologia , Fazendas , Fezes/virologia , Feminino , Gastroenterite Suína Transmissível/mortalidade , Gastroenterite Suína Transmissível/patologia , Grécia , Intestino Delgado/patologia , Masculino , Suínos
17.
Biomed Res Int ; 2018: 5035139, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30533434

RESUMO

Porcine deltacoronavirus (PDCoV) is a newly discovered coronavirus, which belongs to the family Coronaviridae. It causes watery diarrhea, vomiting, and dehydration in newborn piglets. A sensitive RT-PCR method is urgently required to detect PDCoV infection. In this study, we developed and evaluated a conventional RT-PCR assay and a SYBR green-based real-time RT-PCR assay that targeted the PDCoV n gene. Both assays are specific and have the same limit of detection at 2 × 101 copies of RNA molecules per reaction. Eighty-four clinical samples were subjected to both conventional RT-PCR and real-time RT-PCR, and the same positive rate (41.7%) was achieved, which was much higher than the positive rate (26.2%) using a previously described one-step RT-PCR technique. In summary, a conventional RT-PCR technique was successfully established for the detection of PDCoV with the same detection limit as a SYBR green-based real-time RT-PCR assay.


Assuntos
Coronavirus/genética , Coronavirus/isolamento & purificação , Limite de Detecção , Compostos Orgânicos/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Suínos/virologia , Animais , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Doenças dos Suínos/virologia
18.
Indian J Med Res ; 148(3): 329-333, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30425224

RESUMO

Background & objectives: Respiratory tract infections are common among Hajj and Umrah pilgrims which pose a public health risk of spread of respiratory infections. Influenza has been reported from Indian Hajj and Umrah returning pilgrims, but data on other respiratory pathogens are sparse in India. Here we report the presence of common respiratory viral pathogens in returning Hajj and Umrah pilgrims suffering from acute respiratory illness (ARI) in 2014-2015. Methods: Respiratory specimens (nasopharyngeal and throat swabs) were collected from 300 consenting pilgrims with ARI in the past one week and tested for influenza and Middle East Respiratory Syndrome coronavirus (MERS-CoV) and other respiratory viruses using in-house standardized quantitative real-time reverse-transcription polymerase chain reaction. Clinical features among the pathogen positive and negative patients were compared. The patients received symptomatic treatment and antivirals where appropriate and were followed telephonically to collect data on illness outcome. Results: Ninety seven (32.3%) of the 300 participants were tested positive for any virus, most common being influenza viruses (n=33, 11%). Other respiratory viruses that were detected included human coronaviruses [n=26, 8.7%; OC43 (n=19, 6.3%) and C229E (n=7, 2.3%)], rhinovirus (n=20, 6%), adenoviruses (n=8, 2.6%), parainfluenza viruses (n=7, 2.3%), respiratory syncytial virus (n=3, 1%) and bocaviruses (n=2, 0.6%). Clinical features observed in pathogen positive and pathogen negative patients did not differ significantly. Eighteen influenza positive patients were treated with oseltamivir. Interpretation & conclusions: Pilgrims returning from mass gatherings are often afflicted with respiratory pathogens with a potential to facilitate transmission of respiratory pathogens across international borders. The study reinforces the need for better infection prevention and control measures such as vaccination, health education on cough etiquette and hand hygiene.


Assuntos
Coronavirus/isolamento & purificação , Transmissão de Doença Infecciosa , Orthomyxoviridae/isolamento & purificação , Vírus Sincicial Respiratório Humano/isolamento & purificação , Infecções Respiratórias , Adulto , Controle de Doenças Transmissíveis/métodos , Transmissão de Doença Infecciosa/prevenção & controle , Transmissão de Doença Infecciosa/estatística & dados numéricos , Feminino , Humanos , Índia/epidemiologia , Masculino , Pessoa de Meia-Idade , Saúde Pública/métodos , Saúde Pública/estatística & dados numéricos , Missões Religiosas/estatística & dados numéricos , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Viagem/estatística & dados numéricos
19.
Res Vet Sci ; 120: 63-69, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30265872

RESUMO

Porcine epidemic diarrhea virus (PEDV) and porcine deltacoronavirus (PDCoV) are epizootic swine viruses. To detect and study the evolution of PEDV and PDCoV in central China (Shanxi, Henan, Hubei province), 70 clinical intestinal and fecal samples from piglets with severe watery diarrhea during August 2015 and June 2016 were collected, tested and analyzed. PEDV was more frequently detected by PCR than PDCoV. Phylogenetic analysis of S genes showed that the 10 PEDV strains from this study clustered into G2a (n = 7) and G2b (n = 3) groups. Additionally, the three G2b strains (PEDV S2△) contained the same specific 3 nt deletion in S2 as other reference strains in G2b. Interestingly, complete genome analysis indicated that CH/hubei/2016 was closer to the US INDEL strain and G2a group. CH/hubei/2016 had one recombination event in S2 gene which may have resulted from AH2012-12 (from G2b group) and CH-ZMDZY-11 (from G2a group). Furthermore, 10 purifying selection sites in S gene indicated an adaptive evolution of PEDV in central China swine herds. These results suggested that Pandemic G2a and G2b are predominant PEDV genotype circulating in central China. In addition, the deletion and recombination identified in S gene suggested PEDV strains of central exhibited an evolutionary variety. However, whether these changes affect the pathogenicity and antigenicity of wild PEDV is unknown and is worth for further investigation.


Assuntos
Infecções por Coronavirus/veterinária , Coronavirus/genética , Evolução Molecular , Doenças dos Suínos/virologia , Animais , China/epidemiologia , Coronavirus/isolamento & purificação , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Diarreia/veterinária , Fezes/virologia , Genótipo , Filogenia , Reação em Cadeia da Polimerase , Suínos , Doenças dos Suínos/epidemiologia
20.
Emerg Microbes Infect ; 7(1): 154, 2018 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-30209269

RESUMO

SARS coronavirus (SARS-CoV), the causative agent of the large SARS outbreak in 2003, originated in bats. Many SARS-like coronaviruses (SL-CoVs) have been detected in bats, particularly those that reside in China, Europe, and Africa. To further understand the evolutionary relationship between SARS-CoV and its reservoirs, 334 bats were collected from Zhoushan city, Zhejiang province, China, between 2015 and 2017. PCR amplification of the conserved coronaviral protein RdRp detected coronaviruses in 26.65% of bats belonging to this region, and this number was influenced by seasonal changes. Full genomic analyses of the two new SL-CoVs from Zhoushan (ZXC21 and ZC45) showed that their genomes were 29,732 nucleotides (nt) and 29,802 nt in length, respectively, with 13 open reading frames (ORFs). These results revealed 81% shared nucleotide identity with human/civet SARS CoVs, which was more distant than that observed previously for bat SL-CoVs in China. Importantly, using pathogenic tests, we found that the virus can reproduce and cause disease in suckling rats, and further studies showed that the virus-like particles can be observed in the brains of suckling rats by electron microscopy. Thus, this study increased our understanding of the genetic diversity of the SL-CoVs carried by bats and also provided a new perspective to study the possibility of cross-species transmission of SL-CoVs using suckling rats as an animal model.


Assuntos
Quirópteros/virologia , Coronavirus/genética , Coronavirus/patogenicidade , Genoma Viral , Animais , China , Coronavirus/classificação , Coronavirus/isolamento & purificação , Feminino , Masculino , Fases de Leitura Aberta , Filogenia , Ratos , Proteínas Virais/química , Proteínas Virais/genética , Proteínas Virais/metabolismo , Virulência
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