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1.
Cell ; 184(8): 2212-2228.e12, 2021 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-33713620

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can cause acute respiratory disease and multiorgan failure. Finding human host factors that are essential for SARS-CoV-2 infection could facilitate the formulation of treatment strategies. Using a human kidney cell line-HK-2-that is highly susceptible to SARS-CoV-2, we performed a genome-wide RNAi screen and identified virus dependency factors (VDFs), which play regulatory roles in biological pathways linked to clinical manifestations of SARS-CoV-2 infection. We found a role for a secretory form of SARS-CoV-2 receptor, soluble angiotensin converting enzyme 2 (sACE2), in SARS-CoV-2 infection. Further investigation revealed that SARS-CoV-2 exploits receptor-mediated endocytosis through interaction between its spike with sACE2 or sACE2-vasopressin via AT1 or AVPR1B, respectively. Our identification of VDFs and the regulatory effect of sACE2 on SARS-CoV-2 infection shed insight into pathogenesis and cell entry mechanisms of SARS-CoV-2 as well as potential treatment strategies for COVID-19.


Assuntos
Enzima de Conversão de Angiotensina 2/imunologia , Interações entre Hospedeiro e Microrganismos/imunologia , SARS-CoV-2/imunologia , Glicoproteína da Espícula de Coronavírus/imunologia , Vasopressinas/imunologia , Internalização do Vírus , COVID-19/imunologia , COVID-19/virologia , Linhagem Celular , Humanos , Ligação Proteica
3.
Hepatology ; 73(1): 10-22, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-31960460

RESUMO

BACKGROUND AND AIMS: Hepatitis E virus (HEV) variants causing human infection predominantly belong to HEV species A (HEV-A). HEV species C genotype 1 (HEV-C1) circulates in rats and is highly divergent from HEV-A. It was previously considered unable to infect humans, but the first case of human HEV-C1 infection was recently discovered in Hong Kong. The aim of this study is to further describe the features of this zoonosis in Hong Kong. APPROACH AND RESULTS: We conducted a territory-wide prospective screening study for HEV-C1 infection over a 31-month period. Blood samples from 2,860 patients with abnormal liver function (n = 2,201) or immunosuppressive conditions (n = 659) were screened for HEV-C1 RNA. In addition, 186 captured commensal rats were screened for HEV-C1 RNA. Sequences of human-derived and rat-derived HEV-C1 isolates were compared. Epidemiological and clinical features of HEV-C1 infection were analyzed. HEV-C1 RNA was detected in 6/2,201 (0.27%) patients with hepatitis and 1/659 (0.15%) immunocompromised persons. Including the previously reported case, eight HEV-C1 infections were identified, including five in patients who were immunosuppressed. Three patients had acute hepatitis, four had persistent hepatitis, and one had subclinical infection without hepatitis. One patient died of meningoencephalitis, and HEV-C1 was detected in cerebrospinal fluid. HEV-C1 hepatitis was generally milder than HEV-A hepatitis. HEV-C1 RNA was detected in 7/186 (3.76%) rats. One HEV-C1 isolate obtained from a rat captured near the residences of patients was closely related to the major outbreak strain. CONCLUSIONS: HEV-C1 is a cause of hepatitis E in humans in Hong Kong. Immunosuppressed individuals are susceptible to persistent HEV-C1 infection and extrahepatic manifestations. Subclinical HEV-C1 infection threatens blood safety. Tests for HEV-C1 are required in clinical laboratories.


Assuntos
Reservatórios de Doenças/veterinária , Vírus da Hepatite E/genética , Hepatite E/epidemiologia , Hepatite E/transmissão , Idoso , Idoso de 80 Anos ou mais , Animais , Reservatórios de Doenças/virologia , Feminino , Vírus da Hepatite E/classificação , Hepatite Viral Animal/transmissão , Hong Kong/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Filogenia , Estudos Prospectivos , RNA Viral/genética , Ratos , Zoonoses/transmissão , Zoonoses/virologia
4.
Circ J ; 84(11): 2027-2031, 2020 10 23.
Artigo em Inglês | MEDLINE | ID: mdl-32981925

RESUMO

BACKGROUND: SARS-CoV-2 infection is associated with myocardial injury, but there is a paucity of experimental platforms for the condition.Methods and Results:Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) infected by SARS-CoV-2 for 3 days ceased beating and exhibited cytopathogenic changes with reduced viability. Active viral replication was evidenced by an increase in supernatant SARS-CoV-2 and the presence of SARS-CoV-2 nucleocaspid protein within hiPSC-CMs. Expressions of BNP, CXCL1, CXCL2, IL-6, IL-8 and TNF-α were upregulated, while ACE2 was downregulated. CONCLUSIONS: Our hiPSC-CM-based in-vitro SARS-CoV-2 myocarditis model recapitulated the cytopathogenic effects and cytokine/chemokine response. It could be exploited as a drug screening platform.


Assuntos
Betacoronavirus/metabolismo , Infecções por Coronavirus/complicações , Células-Tronco Pluripotentes Induzidas/virologia , Miocardite/complicações , Miócitos Cardíacos/virologia , Pneumonia Viral/complicações , Enzima de Conversão de Angiotensina 2 , Betacoronavirus/genética , COVID-19 , Sobrevivência Celular , Células Cultivadas , Infecções por Coronavirus/metabolismo , Infecções por Coronavirus/virologia , Proteínas do Nucleocapsídeo de Coronavírus , Citocinas/metabolismo , Efeito Citopatogênico Viral , Avaliação Pré-Clínica de Medicamentos/métodos , Humanos , Células-Tronco Pluripotentes Induzidas/metabolismo , Miocardite/metabolismo , Miocardite/virologia , Miócitos Cardíacos/metabolismo , Proteínas do Nucleocapsídeo/metabolismo , Pandemias , Peptidil Dipeptidase A/metabolismo , Fosfoproteínas , Pneumonia Viral/metabolismo , Pneumonia Viral/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , SARS-CoV-2 , Replicação Viral
5.
Int J Mol Sci ; 21(15)2020 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-32751106

RESUMO

To control the COVID-19 pandemic and prevent its resurgence in areas preparing for a return of economic activities, a method for a rapid, simple, and inexpensive point-of-care diagnosis and mass screening is urgently needed. We developed and evaluated a one-step colorimetric reverse-transcriptional loop-mediated isothermal amplification assay (COVID-19-LAMP) for detection of SARS-CoV-2, using SARS-CoV-2 isolate and respiratory samples from patients with COVID-19 (n = 223) and other respiratory virus infections (n = 143). The assay involves simple equipment and techniques and low cost, without the need for expensive qPCR machines, and the result, indicated by color change, is easily interpreted by naked eyes. COVID-19-LAMP can detect SARS-CoV-2 RNA with detection limit of 42 copies/reaction. Of 223 respiratory samples positive for SARS-CoV-2 by qRT-PCR, 212 and 219 were positive by COVID-19-LAMP at 60 and 90 min (sensitivities of 95.07% and 98.21%) respectively, with the highest sensitivities among nasopharyngeal swabs (96.88% and 98.96%), compared to sputum/deep throat saliva samples (94.03% and 97.02%), and throat swab samples (93.33% and 98.33%). None of the 143 samples with other respiratory viruses were positive by COVID-19-LAMP, showing 100% specificity. Samples with higher viral load showed shorter detection time, some as early as 30 min. This inexpensive, highly sensitive and specific COVID-19-LAMP assay can be useful for rapid deployment as mobile diagnostic units to resource-limiting areas for point-of-care diagnosis, and for unlimited high-throughput mass screening at borders to reduce cross-regional transmission.


Assuntos
Betacoronavirus/genética , Colorimetria/métodos , Infecções por Coronavirus/diagnóstico , Programas de Rastreamento/economia , Pneumonia Viral/diagnóstico , RNA Viral/análise , Betacoronavirus/isolamento & purificação , COVID-19 , Colorimetria/economia , Infecções por Coronavirus/virologia , Humanos , Limite de Detecção , Nasofaringe/virologia , Técnicas de Amplificação de Ácido Nucleico/métodos , Pandemias , Pneumonia Viral/virologia , Sistemas Automatizados de Assistência Junto ao Leito , RNA Viral/metabolismo , SARS-CoV-2 , Carga Viral
6.
Int J Mol Sci ; 20(23)2019 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-31779252

RESUMO

Enterovirus A71 (EV-A71) and coxsackievirus A16 (CV-A16) are the most common causes of hand, foot, and mouth disease. Severe EV-A71 and CV-A16 infections may be associated with life-threatening complications. However, the pathogenic mechanisms underlying these severe clinical and pathological features remain incompletely understood. Lipids are known to play critical roles in multiple stages of the virus replication cycle. The specific lipid profile induced upon virus infection is required for optimal virus replication. The perturbations in the host cell lipidomic profiles upon enterovirus infection have not been fully characterized. To this end, we performed ultra-high performance liquid chromatography-electrospray ionization-quadrupole-time of flight-mass spectrometry (UPLC-ESI-Q-TOF-MS)-based lipidomics to characterize the change in host lipidome upon EV-A71 and CV-A16 infections. Our results revealed that 47 lipids within 11 lipid classes were significantly perturbed after EV-A71 and CV-A16 infection. Four polyunsaturated fatty acids (PUFAs), namely, arachidonic acid (AA), docosahexaenoic acid (DHA), docosapentaenoic acid (DPA), and eicosapentaenoic acid (EPA), were consistently upregulated upon EV-A71 and CV-A16 infection. Importantly, exogenously supplying three of these four PUFAs, including AA, DHA, and EPA, in cell cultures significantly reduced EV-A71 and CV-A16 replication. Taken together, our results suggested that enteroviruses might specifically modulate the host lipid pathways for optimal virus replication. Excessive exogenous addition of lipids that disrupted this delicate homeostatic state could prevent efficient viral replication. Precise manipulation of the host lipid profile might be a potential host-targeting antiviral strategy for enterovirus infection.


Assuntos
Enterovirus Humano A/patogenicidade , Infecções por Enterovirus/metabolismo , Lipidômica/métodos , Linhagem Celular , Cromatografia Líquida de Alta Pressão , Enterovirus Humano A/classificação , Infecções por Enterovirus/virologia , Homeostase , Humanos , Análise de Componente Principal , Espectrometria de Massas por Ionização por Electrospray , Replicação Viral
7.
J Clin Microbiol ; 53(11): 3639-45, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26311865

RESUMO

Our case series showed that uncomplicated Yarrowia lipolytica fungemia might be treated with catheter removal alone. The Vitek 2 YST identification (ID) card system, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), and internal transcribed spacer and 25S nuclear ribosomal DNA (nrDNA) gene sequencing provided reliable identification. All isolates had low MICs to voriconazole, echinocandins, and amphotericin B.


Assuntos
Antifúngicos/uso terapêutico , Infecções Relacionadas a Cateter/diagnóstico , DNA Espaçador Ribossômico/genética , Fungemia/diagnóstico , Fungemia/tratamento farmacológico , Yarrowia/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Anfotericina B/farmacologia , Sequência de Bases , Infecções Relacionadas a Cateter/tratamento farmacológico , Infecções Relacionadas a Cateter/microbiologia , Cateterismo Venoso Central/efeitos adversos , Cateterismo Periférico/efeitos adversos , Pré-Escolar , Equinocandinas/farmacologia , Feminino , Fungemia/microbiologia , Hospitalização , Humanos , Lactente , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Dados de Sequência Molecular , Estudos Prospectivos , Análise de Sequência de DNA , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Voriconazol/farmacologia , Yarrowia/efeitos dos fármacos
8.
J Clin Microbiol ; 53(8): 2722-6, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26019210

RESUMO

Based on findings in small RNA-sequencing (Seq) data analysis, we developed highly sensitive and specific real-time reverse transcription (RT)-PCR assays with locked nucleic acid probes targeting the abundantly expressed leader sequences of Middle East respiratory syndrome coronavirus (MERS-CoV) and other human coronaviruses. Analytical and clinical evaluations showed their noninferiority to a commercial multiplex PCR test for the detection of these coronaviruses.


Assuntos
Infecções por Coronavirus/diagnóstico , Coronavirus/classificação , Coronavirus/isolamento & purificação , Sondas de Oligonucleotídeos/genética , Oligonucleotídeos/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Criança , Pré-Escolar , Coronavirus/genética , Infecções por Coronavirus/virologia , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Adulto Jovem
9.
J Infect Dis ; 207(11): 1743-52, 2013 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-23532101

RESUMO

The emerging novel human betacoronavirus 2c EMC/2012 (HCoV-EMC) was recently isolated from patients with severe pneumonia and renal failure and was associated with an unexplained high crude fatality rate of 56%. We performed a cell line susceptibility study with 28 cell lines. HCoV-EMC was found to infect the human respiratory tract (polarized airway epithelium cell line Calu-3, embryonic fibroblast cell line HFL, and lung adenocarcinoma cell line A549), kidney (embryonic kidney cell line HEK), intestinal tract (colorectal adenocarcinoma cell line Caco-2), liver cells (hepatocellular carcinoma cell line Huh-7), and histiocytes (malignant histiocytoma cell line His-1), as evident by detection of high or increasing viral load in culture supernatants, detection of viral nucleoprotein expression by immunostaining, and/or detection of cytopathic effects. Although an infected human neuronal cell line (NT2) and infected monocyte and T lymphocyte cell lines (THP-1, U937, and H9) had increased viral loads, their relatively lower viral production corroborated with absent nucleoprotein expression and cytopathic effects. This range of human tissue tropism is broader than that for all other HCoVs, including SARS coronavirus, HCoV-OC43, HCoV-HKU1, HCoV-229E, and HCoV-NL63, which may explain the high mortality associated with this disease. A recent cell line susceptibility study showed that HCoV-EMC can infect primate, porcine, and bat cells and therefore may jump interspecies barriers. We found that HCoV-EMC can also infect civet lung fibroblast and rabbit kidney cell lines. These findings have important implications for the diagnosis, pathogenesis, and transmission of HCoV-EMC.


Assuntos
Infecções por Coronavirus/patologia , Infecções por Coronavirus/virologia , Coronavirus/patogenicidade , Tropismo Viral , Linhagem Celular , Coronavirus/fisiologia , Humanos , Carga Viral , Cultura de Vírus , Replicação Viral
10.
Emerg Microbes Infect ; 13(1): 2373317, 2024 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38934251

RESUMO

Tsukamurella, a group of multi-drug resistant, Gram-positive, aerobic, and partially acid-fast bacteria, are emerging causes of bacterial conjunctivitis and keratitis. However, the pathogenesis of Tsukamurella keratitis is largely unknown. To address this, we used New Zealand White rabbits to develop the first eye infection model and conducted in vitro tests to study the pathogenesis mechanisms of Tsukamurella. There is increasing evidence that biofilms play a significant role in ocular infections, leading us to hypothesize that biofilm formation is crucial for effective Tsukamurella infection. In order to look for potential candidate genes which are important in biofilm formation and Tsukamurella keratitis. We performed genome sequencing of two ocular isolates, T. pulmonis-PW1004 and T. tyrosinosolvens-PW899, to identify potential virulence factors. Through in vitro and in vivo studies, we characterized their biological roles in mediating Tsukamurella keratitis. Our findings confirmed that Tsukamurella is an ocular pathogen by fulfilling Koch's postulates, and using genome sequence data, we identified tmytC, encoding a mycolyltransferase, as a crucial gene in biofilm formation and causing Tsukamurella keratitis in the rabbit model. This is the first report demonstrating the novel role of mycolyltransferase in causing ocular infections. Overall, our findings contribute to a better understanding of Tsukamurella pathogenesis and provide a potential target for treatment. Specific inhibitors targeting TmytC could serve as an effective treatment option for Tsukamurella infections.


Assuntos
Biofilmes , Modelos Animais de Doenças , Ceratite , Biofilmes/crescimento & desenvolvimento , Animais , Coelhos , Ceratite/microbiologia , Fatores de Virulência/genética , Fatores de Virulência/metabolismo , Infecções por Actinomycetales/microbiologia , Infecções por Actinomycetales/veterinária , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sequenciamento Completo do Genoma , Infecções Oculares Bacterianas/microbiologia , Genoma Bacteriano , Humanos
11.
J Formos Med Assoc ; 112(7): 372-81, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23883791

RESUMO

A novel lineage C betacoronavirus, originally named human coronavirus EMC/2012 (HCoV-EMC) and recently renamed Middle East respiratory syndrome coronavirus (MERS-CoV), that is phylogenetically closely related to Tylonycteris bat coronavirus HKU4 and Pipistrellus bat coronavirus HKU5, which we discovered in 2007 from bats in Hong Kong, has recently emerged in the Middle East to cause a severe acute respiratory syndrome (SARS)-like infection in humans. The first laboratory-confirmed case, which involved a 60-year-old man from Bisha, the Kingdom of Saudi Arabia (KSA), who died of rapidly progressive community-acquired pneumonia and acute renal failure, was announced by the World Health Organization (WHO) on September 23, 2012. Since then, a total of 70 cases, including 39 fatalities, have been reported in the Middle East and Europe. Recent clusters involving epidemiologically-linked household contacts and hospital contacts in the Middle East, Europe, and Africa strongly suggested possible human-to-human transmission. Clinical and laboratory research data generated in the past few months have provided new insights into the possible animal reservoirs, transmissibility, and virulence of MERS-CoV, and the optimal laboratory diagnostic options and potential antiviral targets for MERS-CoV-associated infection.


Assuntos
Infecções por Coronaviridae/epidemiologia , Infecções por Coronaviridae/virologia , Coronaviridae , Síndrome Respiratória Aguda Grave/epidemiologia , Síndrome Respiratória Aguda Grave/virologia , Animais , Quirópteros/virologia , Infecções por Coronaviridae/diagnóstico , Infecções por Coronaviridae/tratamento farmacológico , Infecções por Coronaviridae/transmissão , Reservatórios de Doenças/virologia , Descoberta de Drogas , Humanos , Oriente Médio/epidemiologia , Síndrome Respiratória Aguda Grave/tratamento farmacológico
12.
J Formos Med Assoc ; 112(11): 666-75, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24034908

RESUMO

BACKGROUND/PURPOSE: We describe an investigation of an incident of failed sterilization procedure in a dental clinic. We aim to illustrate the principles in performing such investigations and to highlight some of the important checkpoints in sterilization procedures. METHODS: In response to this incident, proper sterilization of all equipment was performed immediately. On-site investigation was conducted by the investigation panel to identify the cause and risks, to coordinate post-exposure management in affected patients, and to make recommendations to prevent similar occurrence of such incidents in the future. RESULTS: The incident was due to a rare lapse of monitoring during the autoclaving cycle. A total of 127 sources and 250 exposed patients were identified within 24 hours of the discovery of the incident for risk assessment and testing for blood-borne viruses, including hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV). A protocol was devised to manage the exposed patients against HBV, HCV, and HIV. Immunization and hyperimmune globulin for hepatitis B, and tetanus toxoids were given to the exposed patients where indicated. Exposed patients were followed-up for 6 months. We came to the decision that dating of instrument packages and signed documentation of each autoclave printout, color change of chemical indicators of each load and daily autoclave performance should be made mandatory with immediate effect. CONCLUSION: Rapid response is extremely crucial in minimizing the impact of this incident and relieving the anxiety of the affected patients. Proper recording and documentation of autoclave cycles and regular auditing should be enforced to prevent similar incidents.


Assuntos
Infecção Hospitalar/prevenção & controle , Clínicas Odontológicas , Esterilização/métodos , Infecção Hospitalar/epidemiologia , Seguimentos , Hong Kong/epidemiologia , Humanos , Incidência , Estudos Retrospectivos , Instrumentos Cirúrgicos
13.
Microbiol Spectr ; 11(6): e0259123, 2023 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-37971222

RESUMO

IMPORTANCE: We report the application of a colorimetric and fluorescent reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay to facilitate mass screening for sarbecoviruses in bats. The assay was evaluated using a total of 838 oral and alimentary samples from bats and demonstrated comparable sensitivity and specificity to quantitative reverse transcription PCR (qRT-PCR), with a simple setup. The addition of SYTO9, a fluorescent nucleic acid stain, also allows for quantitative analysis. The scalability and simplicity of the assay are believed to contribute to improving preparedness for detecting emerging coronaviruses by applying it to field studies and surveillance.


Assuntos
Quirópteros , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave , Animais , Quirópteros/virologia , Técnicas de Diagnóstico Molecular , Técnicas de Amplificação de Ácido Nucleico , Transcrição Reversa
14.
Emerg Microbes Infect ; 11(1): 1900-1909, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-35786393

RESUMO

Drug resistance derived from extracellular vesicles (EVs) is an increasingly important research area but has seldom been described regarding fungal pathogens. Here, we characterized EVs derived from a triazole-resistant but amphotericin B-susceptible strain of Candida auris. Nano- to microgram concentrations of C. auris EVs prepared from both broth and solid agar cultures could robustly increase the yeast's survival against both pure and clinical amphotericin B formulations in a dose-dependent manner, resulting in up to 16-fold changes of minimum inhibitory concentration. Meanwhile, this effect was not observed upon addition of these EVs to C. albicans, nor upon addition of C. albicans EVs to C. auris. No change in susceptibilities was observed upon EV treatment for fluconazole, voriconazole, micafungin, and flucytosine. Mass spectrometry indicated the presence of immunogenic-/drug resistance-implicated proteins in C. auris EVs, including alcohol dehydrogenase 1 as well as C. albicans Mp65-like and Xog1-like proteins in high quantities. Based on these observations, we propose a potential species-specific role for EVs in amphotericin B resistance in C. auris. These observations may provide critical insights into treatment of multidrug-resistant C. auris.


Assuntos
Candidíase , Vesículas Extracelulares , Anfotericina B/farmacologia , Anfotericina B/uso terapêutico , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Candida , Candida albicans , Candida auris , Candidíase/microbiologia , Humanos , Testes de Sensibilidade Microbiana
15.
Front Microbiol ; 12: 795449, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-35095806

RESUMO

Since its first discovery in 1967, human coronavirus OC43 (HCoV-OC43) has been associated with mild self-limiting upper respiratory infections worldwide. Fatal primary pneumonia due to HCoV-OC43 is not frequently described. This study describes a case of fatal primary pneumonia associated with HCoV-OC43 in a 75-year-old patient with good past health. The viral loads of the respiratory tract specimens (bronchoalveolar lavage and endotracheal aspirate) from diagnosis to death were persistently high (3.49 × 106-1.10 × 1010 copies/ml). HCoV-OC43 at a 6.46 × 103 copies/ml level was also detected from his pleural fluid 2 days before his death. Complete genome sequencing and phylogenetic analysis showed that the present HCoV-OC43 forms a distinct cluster with three other HCoV-OC43 from United States, with a bootstrap value of 100% and sharing 99.9% nucleotide identities. Pairwise genetic distance between this cluster and other HCoV-OC43 genotypes ranged from 0.27 ± 0.02% to 1.25 ± 0.01%. In contrast, the lowest pairwise genetic distance between existing HCoV-OC43 genotypes was 0.26 ± 0.02%, suggesting that this cluster constitutes a novel HCoV-OC43 genotype, which we named genotype I. Unlike genotypes D, E, F, G, and H, no recombination event was observed for this novel genotype. Structural modeling revealed that the loop with the S1/S2 cleavage site was four amino acids longer than other HCoV-OC43, making it more exposed and accessible to protease, which may have resulted in its possible hypervirulence.

16.
Front Microbiol ; 12: 739779, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34956112

RESUMO

Since the emergence of Middle East Respiratory Syndrome (MERS) in 2012, there have been a surge in the discovery and evolutionary studies of viruses in dromedaries. Here, we investigated a herd of nine dromedary calves from Umm Al Quwain, the United Arab Emirates that developed respiratory signs. Viral culture of the nasal swabs from the nine calves on Vero cells showed two different types of cytopathic effects (CPEs), suggesting the presence of two different viruses. Three samples showed typical CPEs of Middle East respiratory syndrome (MERS) coronavirus (MERS-CoV) in Vero cells, which was confirmed by partial RdRp gene sequencing. Complete genome sequencing of the three MERS-CoV strains showed that they belonged to clade B3, most closely related to another dromedary MERS-CoV isolate previously detected in Dubai. They also showed evidence of recombination between lineages B4 and B5 in ORF1ab. Another three samples showed non-typical CPEs of MERS-CoV with cell rounding, progressive degeneration, and detachment. Electron microscopy revealed spherical viral particles with peplomers and diameter of about 170nm. High-throughput sequencing and metagenomic analysis showed that the genome organization (3'-N-P-M-F-HN-L-5') was typical of paramyxovirus. They possessed typical genome features similar to other viruses of the genus Respirovirus, including a conserved motif 323FAPGNYALSYAM336 in the N protein, RNA editing sites 5'-717AAAAAAGGG725-3', and 5'-1038AGAAGAAAGAAAGG1051-3' (mRNA sense) in the P gene with multiple polypeptides coding capacity, a nuclear localization signal sequence 245KVGRMYSVEYCKQKIEK261 in the M protein, a conserved sialic acid binding motif 252NRKSCS257 in the HN protein, conserved lengths of the leader (55nt) and trailer (51nt) sequences, total coding percentages (92.6-93.4%), gene-start (AGGANNAAAG), gene-end (NANNANNAAAAA), and trinucleotide intergenic sequences (CTT, mRNA sense). Phylogenetic analysis of their complete genomes showed that they were most closely related to bovine parainfluenza virus 3 (PIV3) genotype C strains. In the phylogenetic tree constructed using the complete L protein, the branch length between dromedary camel PIV3 (DcPIV3) and the nearest node is 0.04, which is >0.03, the definition used for species demarcation in the family Paramyxoviridae. Therefore, we show that DcPIV3 is a novel species of the genus Respirovirus that co-circulated with MERS-CoV in a dromedary herd in the Middle East.

18.
Viruses ; 11(9)2019 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-31480604

RESUMO

Newcastle disease virus (NDV) causes morbidities and mortalities in wild and domestic birds globally. For humans, exposure to infected birds can cause conjunctivitis and influenza-like symptoms. NDV infections in mammals are rarely reported. In this study, using next-generation sequencing, an NDV was identified and isolated from Vero cells inoculated with the nasal swab of an aborted dromedary fetus in Dubai, during the time when an NDV outbreak occurred in a pigeon farm located in close proximity to the dairy camel farm where the mother of the aborted dromedary fetus resided, and there were a lot of pigeons in the camel farm. Genome analysis revealed that the structurally and functionally important features of other NDVs were also present in this dromedary NDV genome. Phylogenetic analysis based on the nucleotide sequences of fusion protein (F), hemagglutinin-neuraminidase protein (HN) and complete polyprotein showed that the virus belonged to sub-genotype VIg of class II NDV and is most closely related to pigeon NDVs in Egypt in the same year. The present study is the first that demonstrated isolation of NDV in dromedaries. Further study is warranted to investigate the relationship between NDV infection and abortion.


Assuntos
Feto Abortado/virologia , Camelus/virologia , Vírus da Doença de Newcastle/isolamento & purificação , Animais , Columbidae/virologia , Egito/epidemiologia , Genoma Viral/genética , Genótipo , Doença de Newcastle/epidemiologia , Doença de Newcastle/virologia , Vírus da Doença de Newcastle/classificação , Vírus da Doença de Newcastle/genética , Filogenia , Proteínas Virais/genética
19.
J Clin Pathol ; 60(2): 202-3, 2007 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-17264245

RESUMO

A case of Ascaris-induced eosinophilic pneumonitis in an HIV-infected patient is described. Owing to his HIV status and the absence of peripheral blood eosinophilia on admission, the initial diagnosis was incorrect until the passage of two worms in his stool. The patient developed eosinophilia subsequently, and examination of his sputum also showed increased eosinophils. The patient gradually improved with inhaled bronchodilators, steroid and mebendazole. As peripheral blood eosinophilia may be transient and the larval migration phase occurs before eggs are present in stool, a high index of suspicion is required in making the diagnosis of Ascaris pneumonitis. Examination of sputum for larvae or increased eosinophils should be performed in patients suspected of having pulmonary infiltrates from endemic areas irrespective of peripheral blood eosinophil counts.


Assuntos
Infecções Oportunistas Relacionadas com a AIDS/diagnóstico , Ascaríase/diagnóstico , Pneumopatias Parasitárias/diagnóstico , Eosinofilia Pulmonar/diagnóstico , Adulto , Animais , Ascaris/isolamento & purificação , Diagnóstico Diferencial , Humanos , Masculino , Eosinofilia Pulmonar/parasitologia , Tuberculose Miliar/diagnóstico
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