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1.
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
3.
Vet Microbiol ; 250: 108853, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32992291

RESUMO

Coronaviruses (CoVs) is showing obvious interspecies transmission, such as the SARS-CoV, MERS-CoV and SARS-CoV-2. Here, the emerging porcine deltacoronavirus (PDCoV) strain, isolated from Shanghai, China, broadly infects porcine, human and chicken cells in vitro. Previously studies by our group and others have confirmed that PDCoV nucleocapsid (N) protein performs an important role in antagonizing retinoic acid-induced gene I-like receptor (RLR) activation. However, the mechanism of PDCoV N protein suppressing porcine type I IFN production remains unclear, especially the downstream of porcine RLR signaling pathway. In the present study, porcine IRF7 (poIRF7) was identified as the interaction protein of PDCoV N protein through LC-MS/MS. The poIRF7 (268-487aa) was the key region of binding PDCoV N protein. Although IRF7 is a conserved functional protein in species, the PDCoV N protein has been confirmed to interact with only poIRF7 and significantly decrease poIRF7-induced type I IFN production, but not human or chicken IRF7. Furthermore, PDCoV N protein can promote poIRF7 degradation via the ubiquitin-proteasome pathway, which directly increased the K6, K11, and K29-linked polyubiquitination of poIRF7. Lysine 359 of poIRF7 was a key site in PDCoV N protein inducing poIRF7 degradation. Taken together, our results reveal a novel mechanism that PDCoV N protein could species-specifically interact with poIRF7 and then promote its degradation to suppress porcine type I IFN production. The novel findings provide a new insight into PDCoV and other zoonotic coronavirus evading the innate immune response of different species.


Assuntos
Coronavirus/química , Fator Regulador 7 de Interferon/imunologia , Interferons/metabolismo , Proteínas do Nucleocapsídeo/imunologia , Animais , Western Blotting , Linhagem Celular , Galinhas , China , Cromatografia Líquida , Coronavirus/classificação , Técnica Indireta de Fluorescência para Anticorpo , Células HEK293 , Humanos , Imunoprecipitação , Interferons/imunologia , Células LLC-PK1 , Filogenia , Plasmídeos , Complexo de Endopeptidases do Proteassoma/metabolismo , Especificidade da Espécie , Suínos , Espectrometria de Massas em Tandem , Ubiquitina/metabolismo , Sequenciamento Completo do Genoma/veterinária
4.
BMC Infect Dis ; 20(1): 639, 2020 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-32867694

RESUMO

BACKGROUND: Dengue fever is an arthropod vector-borne disease transmitted to humans by infected Aedes mosquitoes. Ethiopia has a favorable ecology for arthropods and report high burden of acute febrile illnesses. However, the contribution of arboviral infections to the burden of acute febrile illnesses is barely known. In this study the seropositivity to dengue virus infection and associated risk factors were assessed in Arba Minch districts, southern Ethiopia. METHODS: An institution based cross-sectional study was conducted in a consecutive group of 529 acute febrile patients between May to August 2016. Socio-demographic data, residence place and clinical signs and symptoms were collected using structured questionnaires. Sera were tested for anti-dengue IgG and IgM using Euroimmune indirect immunofluorescent assay. Data analysis was done using SPSS V-20 (IBM Corp, 2012). P-value < 0.05 was taken as statistically significant. RESULT: Seropositivity was 25.1% (133/529) and 8.1% (43/529) for anti- IgG and IgM respectively. CONCLUSION: The high IgM prevalence detected indicate the probability of active transmission with a potential of public health significance that calls for a proactive follow up of the communities in the study area to forecast and avert the risk.


Assuntos
Anticorpos Antivirais/sangue , Vírus da Dengue/imunologia , Dengue/sangue , Dengue/epidemiologia , Febre/sangue , Febre/epidemiologia , Adolescente , Adulto , Animais , Estudos Transversais , Dengue/diagnóstico , Dengue/virologia , Etiópia/epidemiologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , Pessoa de Meia-Idade , Prevalência , Fatores de Risco , Inquéritos e Questionários , Adulto Jovem
5.
Cornea ; 39(12): 1556-1562, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32826650

RESUMO

PURPOSE: To confirm the ocular tropism of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by evaluating the expression of viral entry factors in human ocular tissues using immunohistochemistry. METHODS: Fresh donor corneas and primary explant cultures of corneal, limbal, and conjunctival epithelial cells were evaluated for the expression of viral entry factors. Using immunohistochemistry, the samples were tested for the expression of angiotension-converting enzyme 2 (ACE2), dendritic cell-specific intracellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN), DC-SIGN-related protein (DC-SIGNR), and transmembrane serine protease 2 (TMPRSS2). RESULTS: In total, 5 donor corneas were evaluated for the expression of viral entry factors. In all specimens, both ACE2 and TMPRSS2 were expressed throughout the surface epithelium (corneal, limbal, and conjunctival) and corneal endothelium. In corneal stromal cells, ACE2 was sporadically expressed, whereas TMPRSS2 was absent. DC-SIGN/DC-SIGNR expression varied between donor specimens. Four specimens expressed DC-SIGN/DC-SIGNR in a similar distribution to ACE2, but 1 specimen from a young donor showed no expression of DC-SIGN/DC-SIGNR. ACE2, TMPRSS2, and DC-SIGN/DC-SIGNR were all expressed in the cultured corneal, limbal, and conjunctival epithelial cells. CONCLUSIONS: Both corneal and conjunctival epithelia express ACE2, DC-SIGN/DC-SIGNR, and TMPRSS2, suggesting that the ocular surface is a potential route for the transmission of SARS-CoV-2. The risk of viral transmission with corneal transplantation cannot be ruled out, given the presence of ACE2 in corneal epithelium and endothelium. Cultured corneal, limbal, and conjunctival epithelial cells mimic the expression of viral entry factors in fresh donor tissue and may be useful for future in vitro SARS-CoV-2 infection studies.


Assuntos
Betacoronavirus/fisiologia , Moléculas de Adesão Celular/metabolismo , Túnica Conjuntiva/metabolismo , Epitélio Anterior/metabolismo , Lectinas Tipo C/metabolismo , Peptidil Dipeptidase A/metabolismo , Receptores de Superfície Celular/metabolismo , Serina Endopeptidases/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Células Cultivadas , Túnica Conjuntiva/citologia , Infecções por Coronavirus/imunologia , Células Epiteliais/metabolismo , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Limbo da Córnea/citologia , Masculino , Microscopia de Fluorescência , Pessoa de Meia-Idade , Pandemias , Pneumonia Viral/imunologia , Doadores de Tecidos , Tropismo Viral/fisiologia , Internalização do Vírus , Adulto Jovem
6.
J Infect Dis ; 222(9): 1439-1443, 2020 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-32738141

RESUMO

BACKGROUND: During the coronavirus disease 2019 (COVID-19) pandemic, we detected a new immunofluorescence (IF) pattern in serum autoantibody (autoAb) screening of laboratory-confirmed COVID-19 patients. METHODS: The IF pattern was composed of liver and gastric mucosa staining on rat kidney/liver/stomach sections. RESULTS: We describe 12 patients positive for the cross-reactive antibody, compared with a negative group of 43 hospitalized COVID-19 patients, finding association with either neurologic or thrombotic complications. In sequential pre- and post-COVID-19 serum samples, we confirmed autoAb seroconversion. CONCLUSIONS: Our data indicate that autoAb screening in COVID-19 patients may be easily performed by IF and alert for autoreactive-mediated complications such as thrombotic or neurologic events.


Assuntos
Autoanticorpos/sangue , Betacoronavirus , Infecções por Coronavirus/imunologia , Doenças do Sistema Nervoso/imunologia , Pneumonia Viral/imunologia , Trombose/imunologia , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Animais , Estudos de Casos e Controles , Infecções por Coronavirus/sangue , Infecções por Coronavirus/complicações , Reações Cruzadas/imunologia , Feminino , Ferritinas/sangue , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Masculino , Pessoa de Meia-Idade , Doenças do Sistema Nervoso/virologia , Pandemias , Pneumonia Viral/sangue , Pneumonia Viral/complicações , Ratos , Soroconversão , Testes Sorológicos , Trombose/virologia , Adulto Jovem
7.
Exp Parasitol ; 217: 107963, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32781092

RESUMO

This study analyzed the large-subunit (60S) ribosomal protein L12 of Eimeria tenella (Et60s-RPL12). A full-length cDNA was cloned, and the recombinant protein was expressed in E. coli BL21 and inoculated in rabbits to produce the polyclonal antibody. Quantitative real-time polymerase chain reaction and western blotting were used to analyze the transcription levels of Et60s-RPL12 and translation levels in different developmental stages of E. tenella. The results showed that the mRNA transcription level of Et60s-RPL12 was highest in second-generation merozoites, whereas the translation level was highest in unsporulated oocysts. Indirect immunofluorescence showed that Et60s-RPL12 was localized to the anterior region and surface of sporozoites, except for the two refractile bodies. As the invasion of DF-1 cells progressed, fluorescence intensity was increased, and Et60s-RPL12 was localized to the parasitophorous vacuole membrane (PVM). The secretion assay results using staurosporine indicated that this protein was secreted, but not from micronemes. The role of Et60s-RPL12 in invasion was evaluated in vitro. The results of the invasion assay showed that polyclonal antibody inhibited host cell invasion by the parasite, which reached about 12%. However, the rate of invasion was not correlated with the concentration of IgG.


Assuntos
Eimeria tenella/genética , Proteínas Ribossômicas/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Western Blotting , Ceco/parasitologia , Linhagem Celular , Embrião de Galinha , Galinhas , Biologia Computacional , DNA Complementar/genética , DNA Complementar/metabolismo , Eimeria tenella/química , Eletroforese em Gel de Poliacrilamida , Fezes/parasitologia , Fibroblastos , Técnica Indireta de Fluorescência para Anticorpo , Biossíntese de Proteínas , Coelhos , Reação em Cadeia da Polimerase em Tempo Real , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Ribossômicas/química , Organismos Livres de Patógenos Específicos , Transcrição Genética
8.
J Clin Virol ; 129: 104539, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32679298

RESUMO

BACKGROUND: Serological tests for anti-SARS-CoV-2 antibodies are becoming of great interest to determine seroprevalence in a given population, define previous exposure and identify highly reactive human donors for the generation of convalescent serum as therapeutic. OBJECTIVES: We evaluated the diagnostic performance of the Abbott ARCHITECT SARS-CoV-2 IgG test, a fully automated indirect immunoassay that detects antibodies directed to a recombinant SARS-CoV-2 Nucleocapsid antigen. STUDY DESIGN: Abbott ARCHITECT SARS-CoV-2 IgG immunoassay was compared to an indirect immunofluorescence assay (IFA) on sera from patients with COVID-19 collected at different days after symptoms onset or infected by other human coronaviruses. Comparison with neutralization test was also performed. RESULTS: After 7, 14 and >14 days after onset ARCHITECT was positive on 8.3 %; 61.9 % and 100 % of the tested samples compared to 58.3 %; 85.7 % and 100 % by IFA. The sensitivity was 72 % vs. IFA and 66.7 % vs. a real-time PCR, the specificity was 100 %. On 18 samples with neutralizing activity, 17 were positive by Abbott ARCHITECT SARS-CoV-2 IgG. CONCLUSIONS: In our study, Abbott ARCHITECT SARS-CoV-2 IgG assay showed a satisfactory performance, with a very high specificity. IgG reactivity against SARSCoV-2 N antigen was detectable in all patients by two weeks after symptoms onset. In addition, concordance between this serological response and viral neutralization suggests that a strong humoral response may be predictive of a neutralization activity, regardless of the target antigens. This finding supports the use of this automated serological assay in diagnostic algorithm and public health intervention, especially for high loads of testing.


Assuntos
Anticorpos Antivirais/sangue , Betacoronavirus/imunologia , Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/diagnóstico , Técnica Indireta de Fluorescência para Anticorpo/métodos , Testes de Neutralização/métodos , Pneumonia Viral/diagnóstico , Testes Sorológicos/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Neutralizantes/sangue , Criança , Pré-Escolar , Feminino , Humanos , Imunoglobulina G/sangue , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Pandemias , Sensibilidade e Especificidade , Adulto Jovem
9.
BMC Infect Dis ; 20(1): 458, 2020 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-32605544

RESUMO

BACKGROUND: Anaplasmosis is an emerging acute febrile disease that is caused by a bite of an Anaplasma phagocytophilum-infected hard tick. As for healthy patients, reports on asymptomatic anaplasmosis resulting from such tick bites are rare. CASE PRESENTATION: A 55-year-old female patient visited the hospital with a tick bite in the right infraclavicular region. The tick was suspected to have been on the patient for more than 10 days. PCR and an indirect immunofluorescence assay (IFA) were performed to identify tick-borne infectious diseases. The blood sample collected at admission yielded a positive result in nested PCR targeting Ehrlichia- or Anaplasma-specific genes groEL and ankA. Subsequent sequencing confirmed the presence of A. phagocytophilum, and seroconversion was confirmed by the IFA involving an A. phagocytophilum antigen slide. PCR detected no Rickettsia-specific genes [outer membrane protein A (ompA) or surface cell antigen 1 (sca1)], but seroconversion of spotted fever group (SFG) rickettsiosis was confirmed by an IFA. CONCLUSIONS: This study genetically and serologically confirmed an asymptomatic A. phagocytophilum infection. Although SFG rickettsiosis was not detected genetically, it was detected serologically. These findings indicate the possibility of an asymptomatic coinfection: anaplasmosis plus SFG rickettsiosis. It is, therefore, crucial for clinicians to be aware of potential asymptomatic anaplasmosis following a tick bite.


Assuntos
Anaplasma phagocytophilum/genética , Anaplasma phagocytophilum/imunologia , Anaplasmose/diagnóstico , Infecções Assintomáticas , Coinfecção/diagnóstico , Rickettsia/imunologia , Rickettsiose do Grupo da Febre Maculosa/diagnóstico , Animais , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Picadas de Carrapatos/microbiologia , Carrapatos
10.
BMC Infect Dis ; 20(1): 523, 2020 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-32682398

RESUMO

BACKGROUND: Assessing the burden of rickettsial infections in Ontario, Canada, is challenging since rickettsial infections are not reportable to public health. In the absence of reportable disease data, we assessed the burden of rickettsial infections by examining patient serological data and clinical information. METHODS: Our retrospective, cross-sectional study included patients who had Rickettsia serological testing ordered by their physician, in Ontario, from 2013 to 2018. We tested sera from 2755 non-travel patients for antibodies against spotted fever group rickettsiae (SFGR) and typhus group rickettsiae (TGR) using an indirect immunofluorescence assay (IFA) (positive IgG titers ≥1:64). We classified cases using a sensitive surveillance case definition: confirmed (4-fold increase in IgG titers between acute and convalescent sera with clinical evidence of infection), possible (single positive sera with clinical evidence) and previous rickettsial infection (single positive sera without clinical evidence). We classified cases seropositive for both SFGR and TGR as unspecified Rickettsia infections (URIs). RESULTS: Less than 5% of all patients had paired acute and convalescent sera tested, and of these, we found a single, laboratory-confirmed SFGR case, with a 4-fold increase in IgG titers and evidence of fever, maculopapular rash and headache. There were 45 possible (19 SFGR, 7 TGR, 19 URI) and 580 previous rickettsial infection (183 SFGR, 89 TGR, 308 URI) cases. The rate of positive tests for SFGR, TGR and URI combined (all case classifications) were 4.4 per 100,000 population. For confirmed and possible cases, the most common signs and symptoms were fever, headache, gastrointestinal complaints and maculopapular rash. The odds of having seropositive patients increased annually by 30% (odds ratio = 1.3, 95% confidence interval: 1.23-1.39). CONCLUSIONS: The rates of rickettsial infections in Ontario are difficult to determine. Based on confirmed and possible cases, rates are low, but inclusion of previous rickettsial infection cases would indicate higher rates. We highlight the need for education regarding the importance of testing acute and convalescent sera and consistent completion of the laboratory requisition in confirming rickettsial disease. We suggest further research in Ontario to investigate rickettsial agents in potential vectors and clinical studies employing PCR testing of clinical samples.


Assuntos
Rickettsia typhi/imunologia , Rickettsiose do Grupo da Febre Maculosa/diagnóstico , Rickettsiose do Grupo da Febre Maculosa/epidemiologia , Tifo Endêmico Transmitido por Pulgas/diagnóstico , Tifo Endêmico Transmitido por Pulgas/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antibacterianos/sangue , Criança , Pré-Escolar , Estudos Transversais , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Imunoglobulina G/sangue , Lactente , Masculino , Pessoa de Meia-Idade , Ontário/epidemiologia , Estudos Retrospectivos , Rickettsiose do Grupo da Febre Maculosa/sangue , Rickettsiose do Grupo da Febre Maculosa/microbiologia , Tifo Endêmico Transmitido por Pulgas/sangue , Tifo Endêmico Transmitido por Pulgas/microbiologia , Adulto Jovem
11.
Arch. argent. pediatr ; 118(3): 193-201, jun. 2020. tab, ilus
Artigo em Inglês, Espanhol | LILACS, BINACIS | ID: biblio-1104196

RESUMO

Introducción. La infección respiratoria aguda baja por adenovirus es una importante causa de morbimortalidad en niños. Objetivos: Describir el patrón clínico-epidemiológico y los factores asociados en niños hospitalizados.Métodos. Estudio transversal en niños ingresados por infección respiratoria aguda baja al Hospital de Niños Ricardo Gutiérrez, Buenos Aires, en 2000-2018. El diagnóstico viral se realizó mediante inmunofluorescencia indirecta en secreciones nasofaríngeas. Se compararon características clínico-epidemiológicas de infección por adenovirus con otros virus respiratorios (virus sincicial respiratorio, influenza y parainfluenza). Se utilizó regresión logística múltiple para identificar predictores independientes de infección.Resultados. De 16018 pacientes con infección respiratoria aguda baja, 13545 fueron testeados para virus respiratorios y 6047 (el 45 %) fueron positivos. Adenovirus fue el agente menos frecuente [el 4,4 % (265) de los casos]; presentó una tendencia en descenso durante todo el período estudiado (pico en 2003) y circuló durante todo el año (pico en julio). El 63,8 % eran varones; mediana de edad: 11 meses (rango intercuartílico: 6-20). La presentación clínica más frecuente fue neumonía (el 63 %). El 50 % tenía internaciones previas por causa respiratoria; el 15,6 % eran reingresos; el 58,3 % tenía comorbilidades. El 19,2 % requirió asistencia ventilatoria; el 44 %registró complicaciones. La letalidad fue del 7,7 %. La infección por adenovirus se asoció a edad ≥ 12 meses, sexo masculino, presentación clínica de neumonía, internaciones previas por causas respiratorias y reinternaciones.Conclusiones. Los adenovirus fueron detectados con menor frecuencia que los otros virus respiratorios, aunque presentaron un importante perfil de morbimortalidad


Introduction. Acute lower respiratory tract infection (ALRTI) caused by adenovirus is a major cause of morbidity and mortality in children.Objectives. To describe the clinical and epidemiological pattern and associated factors in hospitalized children.Methods. Cross-sectional study in children admitted due to ALRTI to Hospital de Niños "Ricardo Gutiérrez," in the Autonomous City of Buenos Aires, between 2000 and 2018. Viral diagnosis was done by indirect immunofluorescence in nasopharyngeal secretions. The clinical and epidemiological characteristics of adenovirus infection were compared to other respiratory viruses (respiratory syncytial virus, influenza, and parainfluenza). A multiple logistic regression was done to identify independent predictors of infection.Results. Out of 16 018 patients with ALRTI, 13 545 were tested for respiratory viruses; 6047 (45 %) had a positive result. Adenovirus was the least common agent (4.4 % [265] of cases); it tended towards a reduction over the study period (peak in 2003) and circulated throughout the year (peak in July). In total, 63.8 % of patients were males; median age: 11 months (interquartile range: 6-20). The most common clinical presentation was pneumonia (63 %). Prior admissions due to respiratory conditions were seen in 50 %; 15.6 %were readmissions; 58.3 % had comorbidities. Ventilatory support was required by 19.2 %and complications were recorded in 44 %. The fatality rate was 7.7 %. Adenovirus infection was associated with age ≥ 12 months, male sex, clinical presentation of pneumonia, prior admissions due to respiratory conditions, and readmissions.Conclusions. Adenoviruses were less common than other respiratory viruses, although their morbidity and mortality were important


Assuntos
Humanos , Masculino , Feminino , Lactente , Infecções Respiratórias/epidemiologia , Infecções por Adenoviridae/epidemiologia , Pneumonia , Infecções Respiratórias/virologia , Estudos Epidemiológicos , Criança Hospitalizada , Estudos Transversais , Infecções por Adenoviridae/diagnóstico , Técnica Indireta de Fluorescência para Anticorpo
12.
Rev Bras Parasitol Vet ; 29(2): e000820, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32490893

RESUMO

Toxoplasma gondii is one of the most important protozoa parasites worldwide. Although many seroprevalence studies have been performed in domestic and wild species, data on the cumulative incidence and the spatial distribution of T. gondii in animals are extremely scarce. In the present study, dogs from Botucatu municipality, São Paulo state, were followed for one year and their blood samples were collected on three moments: days 1, 180, and 360. The sera were submitted to the immunofluorescence antibody test (IFAT) to detect IgG antibodies to T. gondii. Age and sex were compared with IFAT results through statistical tests. Spatial analysis was used to detect clusters of seropositive dogs. Among the 350 dogs that were seronegative on day 1, 53 became seropositive in subsequent samplings; thus, cumulative incidence was 15.1% exposed dogs/year. Age and sex were not associated with serological results. The spatial analysis revealed that seropositive dogs were distributed in all the studied areas, with a significant cluster in a zone with poor sanitary conditions and low socioeconomic status. T. gondii is frequent and widely distributed in the urban area of Botucatu, and impoverished areas are possibly associated with high levels of environmental contamination by this parasite.


Assuntos
Anticorpos Antiprotozoários/sangue , Doenças do Cão/epidemiologia , Toxoplasmose Animal/epidemiologia , Animais , Brasil/epidemiologia , Doenças do Cão/diagnóstico , Doenças do Cão/parasitologia , Cães , Feminino , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Incidência , Estudos Longitudinais , Masculino , Estudos Soroepidemiológicos , Análise Espacial , Toxoplasmose Animal/diagnóstico
13.
Exp Parasitol ; 215: 107901, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32525007

RESUMO

Eimeria tenella (E. tenella) has caused severe economic loss in chicken production, especially after the forbidden use of antibiotics in feed. Considering the drug resistant problem caused by misuse of chemoprophylaxis and live oocyst vaccines can affect the productivity of chickens, also it has the risk to reversion of virulence, the development of efficacious, convenient and safe vaccines is still deeply needed. In this study, the EtMic2 protein of E. tenella was anchored on the surface of Lactobacillus plantarum (L. plantarum) NC8 strain. The newly constructed strain was then used to immunize chickens, followed by E. tenella challenge. The results demonstrated that the recombinant strain could provide efficient protection against E. tenella, shown by increased relative body weight gains, percentages of CD4+ and CD8+ T cells, humoral immune response and inflammatory cytokines. In addition, decreased cecum lesion scores and fecal oocyst shedding were also observed during the experiment. In conclusion, this study proves the possibility to use L. plantarum as a vessel to deliver protective antigen to protect chickens against coccidiosis.


Assuntos
Antígeno 12E7/imunologia , Galinhas/parasitologia , Coccidiose/veterinária , Eimeria tenella/imunologia , Doenças das Aves Domésticas/prevenção & controle , Vacinas Protozoárias , Animais , Antígenos de Protozoários/imunologia , Ceco/parasitologia , Coccidiose/economia , Coccidiose/parasitologia , Coccidiose/prevenção & controle , Eimeria tenella/química , Citometria de Fluxo/veterinária , Técnica Indireta de Fluorescência para Anticorpo/veterinária , Imunoglobulina A Secretora/biossíntese , Imunoglobulina G/sangue , Interferon gama/sangue , Interleucina-2/sangue , Intestinos/imunologia , Lactobacillus plantarum/genética , Lactobacillus plantarum/imunologia , Doenças das Aves Domésticas/economia , Doenças das Aves Domésticas/parasitologia , Distribuição Aleatória , Vacinas Sintéticas
14.
Exp Parasitol ; 216: 107942, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32598889

RESUMO

The intracellular protozoan parasite Neospora caninum is incriminated to induce drastic economic losses in both livestock and pet animal industries. Neosporosis is primarily characterized by abortion in cattle and paralytic symptoms in dogs. Because there are no effective treatments or vaccines, diagnosis is critical for Neospora control. Thus, diversification of laboratory tests and specimens used for diagnosis of N. caninum is an essential scientific endeavor to judge and select the most appropriate diagnostic tool. Herein, we provide the first evidence for the utility of urine samples for demonstration of specific antibodies against N. caninum employing an experimentally infected murine model. Specific antibodies to recombinant N. caninum dense granule 7, surface antigen 1, and lysate antigen were assayed using different antibodies-based ELISAs. Urine based IgG ELISA efficiently discriminated between infected mice (acute or chronic infection), and those of non-infected mice. This effect was also noticed for IgG1 and IgG2a suggesting the utility of urine for assessment of T-helper 2- and T-helper 1-mediated immunities, respectively. In addition, reactivity of specific antibody in urine was also confirmed against parasites when indirect fluorescent antibody test was employed. Usefulness of urine as an additional clinical sample for Neospora diagnosis was confirmed via comparison with the relevant control non-infected and infected mouse sera as reference samples. Because of minimum invasiveness and ease of urine collection, this approach might offer new diagnostic opportunities for N. caninum either for the field or research purposes. However, further studies are required to extrapolate this preliminary study and results in the animal species of interest particularly in dogs.


Assuntos
Anticorpos Antiprotozoários/urina , Coccidiose/diagnóstico , Neospora/imunologia , Análise de Variância , Animais , Anticorpos Antiprotozoários/sangue , Chlorocebus aethiops , Coccidiose/imunologia , Coccidiose/parasitologia , Ensaio de Imunoadsorção Enzimática , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Imunoglobulina G/sangue , Imunoglobulina G/urina , Imunoglobulina M/sangue , Imunoglobulina M/urina , Camundongos , Camundongos Endogâmicos BALB C , Neospora/isolamento & purificação , Células Vero
15.
Autoimmun Rev ; 19(8): 102588, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32540447

RESUMO

The commercial tests currently available as second-level tests to detect ANA sub-specificities are generally used independently from the ANA immunofluorescence (IIF) pattern. The aim of this study was to evaluate the efficacy of the use of a customizable pattern-oriented antigenic panel by immunoblot (IB) using the International Consensus on ANA Patterns (ICAP) classification scheme, in order to introduce a novel and updated autoimmune diagnostic flowchart. 710 sera referred for routine ANA testing were selected on the basis of the ANA pattern according to the ICAP nomenclature (nuclear speckled AC-2,4,5; nucleolar AC-8,9,10,29; cytoplasmic speckled AC-18,19,20) and on an IIF titer ≥1:320. They were then assayed by three experimental IB assays using a panel of selected antigens. ICAP-oriented IB detected 515 antibody reactivities vs. 457 of traditional anti-ENA in the nuclear speckled pattern group, 108 vs. 28 in the nucleolar pattern group, and 43 vs. 34 in the cytoplasmic speckled pattern. This pilot study may lead the way for a new approach introducing an ICAP pattern-oriented follow up testing as a valid alternative to the existing standard panels, thus enabling more patients with autoimmune rheumatic disease to be accurately diagnosed.


Assuntos
Algoritmos , Anticorpos Antinucleares , Doenças Autoimunes , Técnicas e Procedimentos Diagnósticos , Immunoblotting , Anticorpos Antinucleares/sangue , Doenças Autoimunes/sangue , Doenças Autoimunes/diagnóstico , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Immunoblotting/normas , Projetos Piloto
16.
Dtsch Med Wochenschr ; 145(17): e96-e100, 2020 08.
Artigo em Alemão | MEDLINE | ID: mdl-32572869

RESUMO

BACKGROUND: In children, the infection with SARS-CoV-2, the cause of COVID-19, tends to be clinically inapparent more often or less severe than in adults. The spread of this infection from children poses a danger to vulnerable adults. Therefore, child care institutions and schools currently are widely closed. METHODS: Since the status of infection tends to be congruent in mothers and their children, we tested 401 mothers of children between 1 and 10 years in the city of Rostock (State of Mecklenburg-Westpomerania, northeast of Germany), for the presence of RNA of SARS-CoV-2 in throat swabs, and of antibodies against SARS-CoV-2 in serum. RESULTS: In none of the mothers tested, RNA of this virus was detected in the throat swab. In the ELISA test, IgG antibodies were positive in one serum sample, IgA antibodies were positive in 11, and borderline in 3 serum samples. All 401 sera were negative in the indirect immunofluorescence test (IIFT) with FITC-labeled IgG, IgA, und IgM antibodies. CONCLUSION: At the time of this study, neither SARS-CoV-2 RNA, nor specific antibodies against SARS-CoV-2 were detectable in the mothers tested in Rostock.


Assuntos
Anticorpos Antivirais/sangue , Betacoronavirus/imunologia , Infecções por Coronavirus/epidemiologia , Pneumonia Viral/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Betacoronavirus/genética , Betacoronavirus/isolamento & purificação , Criança , Pré-Escolar , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Alemanha/epidemiologia , Humanos , Lactente , Masculino , Programas de Rastreamento/métodos , Pessoa de Meia-Idade , Orofaringe/virologia , Pandemias , Pneumonia Viral/diagnóstico , Pneumonia Viral/imunologia , Reação em Cadeia da Polimerase , Prevalência , RNA Viral/análise , Adulto Jovem
17.
Nat Commun ; 11(1): 2420, 2020 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-32415101

RESUMO

Archetypal human pluripotent stem cells (hPSC) are widely considered to be equivalent in developmental status to mouse epiblast stem cells, which correspond to pluripotent cells at a late post-implantation stage of embryogenesis. Heterogeneity within hPSC cultures complicates this interspecies comparison. Here we show that a subpopulation of archetypal hPSC enriched for high self-renewal capacity (ESR) has distinct properties relative to the bulk of the population, including a cell cycle with a very low G1 fraction and a metabolomic profile that reflects a combination of oxidative phosphorylation and glycolysis. ESR cells are pluripotent and capable of differentiation into primordial germ cell-like cells. Global DNA methylation levels in the ESR subpopulation are lower than those in mouse epiblast stem cells. Chromatin accessibility analysis revealed a unique set of open chromatin sites in ESR cells. RNA-seq at the subpopulation and single cell levels shows that, unlike mouse epiblast stem cells, the ESR subset of hPSC displays no lineage priming, and that it can be clearly distinguished from gastrulating and extraembryonic cell populations in the primate embryo. ESR hPSC correspond to an earlier stage of post-implantation development than mouse epiblast stem cells.


Assuntos
Células-Tronco Embrionárias/citologia , Camadas Germinativas/citologia , Células-Tronco Pluripotentes/citologia , Animais , Diferenciação Celular , Cromatina/metabolismo , Metilação de DNA , Epigenoma , Citometria de Fluxo , Técnica Indireta de Fluorescência para Anticorpo , Fase G1 , Camadas Germinativas/metabolismo , Glicólise , Humanos , Sistema de Sinalização das MAP Quinases , Metabolômica , Camundongos , Mitocôndrias/metabolismo , Fosforilação Oxidativa , RNA-Seq , Transdução de Sinais
18.
Z Rheumatol ; 79(5): 447-458, 2020 Jun.
Artigo em Alemão | MEDLINE | ID: mdl-32458159

RESUMO

Testing for antinuclear antibodies (ANA) on human epithelial cell lines (HEp-2) using indirect immunofluorescence (IIF) is central for ruling out or for diagnosing connective tissue diseases and other diseases, such as primary biliary cholangitis and autoimmune hepatitis as well as drug-induced ANA. The comprehensive description of 29 different ANA-IIF patterns by the international consensus of ANA patterns (ICAP) facilitates the harmonization of ANA-IIF diagnostics. Positive ANA tests are frequently observed in healthy individuals and a reason for referral to rheumatologists. In these cases, the detection of anti-DFS70 antibodies can be helpful to exclude systemic autoimmune rheumatic diseases.


Assuntos
Anticorpos Antinucleares/sangue , Doenças Reumáticas , Proteínas Adaptadoras de Transdução de Sinal , Doenças Autoimunes , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Doenças Reumáticas/sangue , Doenças Reumáticas/diagnóstico , Fatores de Transcrição
19.
Rev Soc Bras Med Trop ; 53: e20190433, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32348430

RESUMO

INTRODUCTION: As highly specific molecular biology-based techniques may not be sensitive enough for the diagnosis of American tegumentary leishmaniasis (ATL), clinicians frequently rely on immunological tests before treatment initiation. Hence, the correct combination of diagnostic tests is imperative for ATL diagnosis. We aimed to evaluate the accuracy of the Montenegro (Leishmanin) skin test (MST) in polymerase chain reaction (PCR)-negative patients to accurately detect ATL. METHODS: Patients with a clinical picture compatible with ATL were divided into ATL (confirmed by lesion smear, culture indirect immunofluorescence, and/or histopathology) and no-ATL (diseases that can mimic leishmaniasis) groups. Conventional PCR for the minicircle kDNA of Leishmania was performed, and the MST was carried out for PCR-negative patients. RESULTS: Ninety-nine patients were included in this study, including 79 diagnosed with ATL (6 with mucocutaneous leishmaniasis) and 20 without ATL (no-ATL group). The MST showed a high sensitivity of 90.0% (95% confidence interval [CI] = 69.90-97.21) in PCR-negative patients that was 10% higher than the sensitivity reported in PCR-positive population (79.66%; 95% CI = 67.73-87.96). CONCLUSIONS: One of the most important reasons for PCR negativity among patients with active ATL is the presence of a strong cellular immunological response, especially in chronic and mucocutaneous leishmaniasis. This reinforces the considerable utility of the tests that detect cellular responses against Leishmania antigens such as the MST in PCR-negative patients when the performance in screening situations is questionable.


Assuntos
Testes Intradérmicos/métodos , Leishmania braziliensis/imunologia , Leishmaniose Cutânea/diagnóstico , Adulto , Idoso , Doença Crônica , Estudos Transversais , DNA de Protozoário/genética , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Leishmania braziliensis/genética , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Sensibilidade e Especificidade
20.
BMC Infect Dis ; 20(1): 261, 2020 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-32245372

RESUMO

BACKGROUND: Q fever (Coxiella burnetii infection) has been associated with adverse perinatal outcomes. After investigating the obstetrical importance of Q fever on Reunion island and demonstrating an association between incident Q fever and miscarriage, we conducted a cross-sectional serosurvey to assess the prevalence of Coxiella burnetii infection among parturient women. METHODS: Between January 9 and July 24, 2014, within the level-4 maternity of Saint Pierre hospital and the level-1 maternity of Le Tampon, we proposed to screen all parturient women for Coxiella burnetii serology. Seropositivity was defined using indirect immunofluorescence for a dilution of phase 2 IgG titre ≥1:64. Further dilutions were chosen to discriminate recent or active infections from past or prevalent infections (< 1:128) and classify these as either possible (1:128), or probable (≥1:256). Recurrent miscarriage, stillbirth, preterm birth, small-for-gestational as well as a composite outcome of these adverse pregnancy outcomes were compared according to seropositivity using bivariate analysis or propensity score matching of seropositive and seronegative women on confounding factors. RESULTS: Among 1112 parturient women screened for Q fever over this 7-month period, 203 (18.3%) were seropositive. Overall weighted seroprevalence was of 20.1% (95%CI, 17.7-22.5%). Weighted seroprevalence of probable infections was 4.7% (95%CI 3.4-5.9%), while > 90% of positive serologies corresponded to past infections or false positives. Seropositivity was associated with none of the abovementioned adverse perinatal outcomes, whether in unpaired or matched analyses on propensity score. CONCLUSION: The magnitude and the pattern of seroprevalence suggest that Q fever is endemic on Reunion island. In this context, we found no significant contribution of prevalent Coxiella burnetii infection to adverse pregnancy outcomes. Although reassuring, these data put in our endemic context, with a previously demonstrated increased risk of incident Q fever associated miscarriage, encourage us to protect pregnant women against the risk of new infection, periconceptional or early in pregnancy.


Assuntos
Coxiella burnetii/imunologia , Parto , Febre Q/epidemiologia , Aborto Espontâneo/microbiologia , Adulto , Anticorpos Antibacterianos/sangue , Coxiella burnetii/isolamento & purificação , Estudos Transversais , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Pessoa de Meia-Idade , Gravidez , Nascimento Prematuro/microbiologia , Prevalência , Reunião/epidemiologia , Estudos Soroepidemiológicos , Natimorto , Adulto Jovem
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