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1.
Recurso na Internet em Espanhol | LIS - Localizador de Informação em Saúde | ID: lis-LISBR1.1-46990

RESUMO

Los coronavirus son una familia de virus que causan enfermedades que van desde el resfriado común hasta enfermedades respiratorias más graves, circulan entre humanos y animales. A veces, los coronavirus que infectan a los animales pueden evolucionar, transmitirse a las personas y convertirse en una nueva cepa de coronavirus capaz de provocar enfermedades en los seres humanos, tal y como sucedió con el Síndrome Respiratorio Agudo Severo (SARS), en Asia en febrero de 2003 y, el Síndrome Respiratorio de Oriente Medio (MERS-CoV), que fue detectado por primera vez en Arabia Saudita en 2012.


Assuntos
Betacoronavirus/patogenicidade , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/diagnóstico , Doença Relacionada a Viagens , Controle Sanitário de Viajantes , Infecções por Coronavirus/transmissão
2.
Washington; OPS; 01/02/2020. 5 p.
Não convencional em Inglês, Espanhol | LILACS | ID: biblio-1049524

RESUMO

En las directrices de laboratorio para la detección y diagnóstico de la infección con el nuevo coronovirus 2019 la Organización Panamericana de la Salud/Organización Mundial de la Salud (OPS/OMS) recomienda a los Estados Miembros garantizar su identificación oportuna, el envío de las muestras a laboratorios nacionales y de referencia y la implementación del protocolo de detección molecular para 2019-nCoV, según la capacidad del laboratorio.


Assuntos
Humanos , Infecções por Coronavirus/diagnóstico , Betacoronavirus/isolamento & purificação , Técnicas de Diagnóstico Molecular , Reação em Cadeia da Polimerase em Tempo Real
3.
Rio de Janeiro; SES/RJ; 28 jan. 2020. 4 p. (Nota Técnica-SVS/SES-RJ, 01).
Monografia em Português | LILACS, CONASS, SES-RJ | ID: biblio-1048858

RESUMO

Os Coronavírus (CoV) compõem uma grande família de vírus, conhecidos desde meados da década de 1960. Podem causar desde um resfriado comum até síndromes respiratórias graves, como a síndrome respiratória aguda grave (SARS - Severe Acute Respiratory Syndrome) e a síndrome respiratória do Oriente Médio (MERS - Middle East Respiratory Syndrome). Os casos agora identificados estão relacionados a uma nova variante do Coronavírus, denominada 2019-nCoV, até então não identificada em humanos.


Assuntos
Humanos , Infecções por Coronavirus/epidemiologia , Betacoronavirus , Brasil/epidemiologia , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/prevenção & controle
4.
Vet Microbiol ; 236: 108387, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31500721

RESUMO

Given the highly contagious and acute nature of porcine epidemic diarrhea (PED), especially in piglets, there is an urgent need for the development of rapid and sensitive diagnostic assays. The diagnostic potentials of specific porcine epidemic diarrhea virus (PEDV) accessory and nonstructural proteins, if any, have not yet been investigated. In order to determine and compare which of the viral proteins may be useful as diagnostic antigens, whole virus (WV) particles and a panel of structural and nonstructural PEDV proteins [spike subunit 1 (S1), the C-terminal part of ORF3 (ORF3C), envelope (E), nonstructural protein 1 (Nsp1), Nsp2, Ac (acidic domain of Nsp3), and ADRP (ADP-ribose-1-monophosphatase domain of Nsp3), expressed individually in bacterial and/or mammalian cells] were tested for reactivity with sera from PEDV-infected pigs by ELISA and/or western blot analysis. According to western blots, serum antibody interactions with the S1 protein were relatively more sensitive and specific than ORF3C, E and Ac. Furthermore, a total of 851 serum samples from diarrheal pigs of different ages were analyzed by ELISA, with most showing immune-reactivity towards the WV, S1, ORF3C, and E proteins. The earliest IgG antibody response was observed in the one-week-old piglets, with similar antibody ontogeny and patterns of seroconversion for S1, ORF3C, E, and WV antigens. In addition, the pattern of neutralizing antibody was more similar to that of IgA in weaning piglets after PEDV infection. Collectively, these data provide more reliable information on the host immune response to different viral proteins, which will be useful for development of novel serological assays and for design of vaccines that better stimulate protective immunity.


Assuntos
Infecções por Coronavirus/veterinária , Vírus da Diarreia Epidêmica Suína/metabolismo , Doenças dos Suínos/virologia , Proteínas Virais/metabolismo , Animais , Anticorpos Antivirais/sangue , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/virologia , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Vírus da Diarreia Epidêmica Suína/imunologia , Suínos , Doenças dos Suínos/diagnóstico , Doenças dos Suínos/imunologia , Células Vero , Proteínas Virais/imunologia
5.
Internist (Berl) ; 60(11): 1136-1145, 2019 Nov.
Artigo em Alemão | MEDLINE | ID: mdl-31455974

RESUMO

BACKGROUND: There are six human pathogenic coronaviruses (CoV), which mainly cause infections of the respiratory system. In everyday clinical practice, it is helpful to know the relevance and characteristics of these pathogens. OBJECTIVE: To present the epidemiology, clinical picture and differences of human pathogenic CoV and to provide information on the diagnostics and treatment of patients suspected of having CoV infections. MATERIAL AND METHODS: Selective literature search, presentation of results and discussion of fundamental works and expert recommendations, including publications by the World Health Organization (WHO), the European Centre for Disease Prevention and Control (ECDC) and the Robert Koch Institute. RESULTS: The four endemic human CoVs (HCoV-NL63, HCoV-229E, HCoV-OC43 and HCoV-HKU1) mainly cause mild respiratory tract infections. In addition to these four endemic HCoV, the two epidemic CoV, severe acute respiratory syndrome (SARS)-CoV and Middle East respiratory syndrome (MERS)-CoV can cause severe pneumonia. The SARS-CoV has not been detected in humans in the last 15 years and MERS-CoV has been circulating mainly on the Arabian Peninsula since 2012; however, neither a specific treatment nor approved vaccines exist for any of the six human pathogenic CoVs. CONCLUSION: All six human CoVs can be diagnosed using RT-PCR on respiratory specimens but this is rarely necessary for the four endemic strains. In current clinical practice SARS-CoV has no importance as it has not been detected in humans for 15 years; however, a possible MERS-CoV infection should be taken into account in patients with typical symptoms and travel history to endemic regions. In this case, rapid diagnostic and general hygiene practices are important to prevent further transmission.


Assuntos
Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/prevenção & controle , Coronavirus , Infecções Respiratórias , Síndrome Respiratória Aguda Grave/diagnóstico , Betacoronavirus , Coronavirus Humano 229E , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Coronavirus Humano NL63 , Coronavirus Humano OC43 , Humanos , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Síndrome Respiratória Aguda Grave/epidemiologia , Síndrome Respiratória Aguda Grave/prevenção & controle , Síndrome Respiratória Aguda Grave/virologia
6.
Artigo em Inglês | MEDLINE | ID: mdl-31311073

RESUMO

Middle East respiratory syndrome coronavirus (MERS-CoV) is a great public health concern globally. Although 83% of the globally confirmed cases have emerged in Saudi Arabia, the spatiotemporal clustering of MERS-CoV incidence has not been investigated. This study analysed the spatiotemporal patterns and clusters of laboratory-confirmed MERS-CoV cases reported in Saudi Arabia between June 2012 and March 2019. Temporal, seasonal, spatial and spatiotemporal cluster analyses were performed using Kulldorff's spatial scan statistics to determine the time period and geographical areas with the highest MERS-CoV infection risk. A strongly significant temporal cluster for MERS-CoV infection risk was identified between April 5 and May 24, 2014. Most MERS-CoV infections occurred during the spring season (41.88%), with April and May showing significant seasonal clusters. Wadi Addawasir showed a high-risk spatial cluster for MERS-CoV infection. The most likely high-risk MERS-CoV annual spatiotemporal clusters were identified for a group of cities (n = 10) in Riyadh province between 2014 and 2016. A monthly spatiotemporal cluster included Jeddah, Makkah and Taif cities, with the most likely high-risk MERS-CoV infection cluster occurring between April and May 2014. Significant spatiotemporal clusters of MERS-CoV incidence were identified in Saudi Arabia. The findings are relevant to control the spread of the disease. This study provides preliminary risk assessments for the further investigation of the environmental risk factors associated with MERS-CoV clusters.


Assuntos
Infecções por Coronavirus/epidemiologia , Coronavírus da Síndrome Respiratória do Oriente Médio , Análise por Conglomerados , Infecções por Coronavirus/diagnóstico , Humanos , Incidência , Estudos Retrospectivos , Arábia Saudita/epidemiologia , Estações do Ano , Análise Espaço-Temporal
7.
Saudi Med J ; 40(7): 714-720, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31287133

RESUMO

OBJECTIVES:   To assess virus knowledge among dentists in Saudi Arabia and to identify factors associated with recommended management practices of patients. Method: A structured questionnaire was distributed to dentists in major Saudi cities between September 2016 and December 2017. The questionnaire investigated participants' knowledge about Middle East Respiratory Syndrome Coronavirus (MERS-CoV) transmission, consequences, patient identification and history taking practices. Data was collected using paper-based questionnaires or an online link sent to dentists registered with Saudi Dental Society nationwide. The analysis was carried using Statistical Package for Social Sciences for Windows, version 22.0 (IBM Corp., Armonk, NY, USA) logistic regression, odds ratio and confidence intervals to identify the relationship between variables. RESULTS: A total of 423 dentists responded the paper-based questionnaire. Overall the knowledge was good with gaps in history taking practices. Best management practices of MERS-CoV patients were significantly higher among dentists with better knowledge of virus transmission (odd ration [OR]=1.16, p less than 0.0001), patients' identification (OR=1.40, p less than 0.0001) and those knowing that corona infection can be fatal (OR= 2.44, p=0.02). CONCLUSION: Best management practices depends on correct patient identification. Educational campaigns should target dentists, given the unique nature of dental practice.


Assuntos
Competência Clínica , Infecções por Coronavirus/prevenção & controle , Odontólogos , Adulto , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/transmissão , Estudos Transversais , Feminino , Humanos , Controle de Infecções Dentárias , Masculino , Anamnese , Coronavírus da Síndrome Respiratória do Oriente Médio , Isolamento de Pacientes , Arábia Saudita , Inquéritos e Questionários
8.
Poult Sci ; 98(11): 5401-5411, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31265112

RESUMO

Infectious bronchitis virus (IBV) and Newcastle disease virus (NDV) are both important viruses seriously affecting poultry industry worldwide. In this study, reverse-transcription LAMP (RT-LAMP) was combined with lateral flow dipstick (LFD) forming a novel detection tool which could simultaneously detect IBV and NDV visually. Primers targeted the 5'-untranslated region (5'-UTR) of IBV genome and the conserved region of NDV large polymerase gene (LP). The specificity and sensitivity of this multiple reverse transcription-LAMP-LFD (mRT-LAMP-LFD) assay were compared with those of conventional RT-PCR, nested RT-PCR (nRT-PCR), quantification RT-PCR (qRT-PCR), and RT-LAMP monitored by electrophoresis. No non-specific amplifications were observed when the assays were tested with unrelated viruses. According to the sensitivity study, when detecting IBV or NDV alone, the lowest detection limits of mRT-LAMP-LFD were 100.8 IBV RNA copies/reaction and 100.7 NDV RNA copies/reaction. Furthermore, when detecting IBV and NDV simultaneously, the lowest detection limit was the same as that of the single detection assays. In the clinical sample study, mRT-LAMP-LFD performed the best among these assays. When tested with IBV or NDV single infected samples, the mean detection rates were 98.65% and 97.25%, respectively. In the IBV and NDV co-infected sample study, the mean detection rates of IBV and NDV were both 95%. This study showed that mRT-LAMP-LFD was a promising qualitative detection tool suitable for field single or multiple IBV and NDV detection.


Assuntos
Galinhas , Cromatografia/veterinária , Infecções por Coronavirus/veterinária , Doença de Newcastle/diagnóstico , Técnicas de Amplificação de Ácido Nucleico/veterinária , Doenças das Aves Domésticas/diagnóstico , Animais , China , Cromatografia/métodos , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/virologia , Vírus da Bronquite Infecciosa/isolamento & purificação , Doença de Newcastle/virologia , Vírus da Doença de Newcastle/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/métodos , Doenças das Aves Domésticas/virologia
9.
Public Health ; 173: 29-32, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31252151

RESUMO

OBJECTIVE: We aimed to evaluate the level of knowledge of Middle East respiratory syndrome coronavirus (MERS-CoV) among Hajj pilgrims before and after an education health programme during international vaccine consultations in France. STUDY DESIGN: A cross-sectional study was performed in the consultation for travel medicine and international vaccination in Reims University Hospital between July 2014 and October 2015. METHODS: Consecutive adults (>18 years old) who attended for pre-Hajj meningococcal vaccination were eligible to complete an anonymous questionnaire with closed answers to evaluate their level of knowledge about MERS-CoV. To evaluate the effectiveness of the information given during the consultation, the same questionnaire was completed by the Hajj pilgrim before and after the consultation, where the information about MERS-CoV was provided. RESULTS: Among 82 Hajj pilgrim adults enrolled in the study, less than 25% were aware of the routes of transmission, symptoms and preventive behaviours to adopt abroad or in case of fever. Pilgrims had a higher rate of correct responses on each question at the time they completed the second questionnaire, as compared with the first, with 11 of 13 questions answered significantly better after delivery of educational information about MERS-CoV. However, although the rate of correct answers to the questions about routes of transmission, symptoms, preventive behaviours to adopt in case of fever and time delay between return and potential MERS-CoV occurrence increased significantly after receiving the information, the rates remained below 50%. CONCLUSION: Information given during travel consultations significantly increases the general level of knowledge, but not enough to achieve epidemic control.


Assuntos
Infecções por Coronavirus/prevenção & controle , Educação em Saúde/organização & administração , Conhecimentos, Atitudes e Prática em Saúde , Promoção da Saúde , Coronavírus da Síndrome Respiratória do Oriente Médio , Avaliação de Programas e Projetos de Saúde/métodos , Adolescente , Adulto , Infecções por Coronavirus/diagnóstico , Estudos Transversais , Feminino , França , Humanos , Masculino , Encaminhamento e Consulta , Infecções Respiratórias/prevenção & controle , Infecções Respiratórias/virologia , Inquéritos e Questionários , Viagem/psicologia , Vacinação
10.
BMC Vet Res ; 15(1): 161, 2019 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-31118011

RESUMO

BACKGROUND: Bovine coronavirus (BCV) is associated with respiratory infections in cattle of all ages; however, a temporal study to evaluate the effect of BCV immunity on virus shedding and bovine respiratory disease (BRD) incidence in pre-weaned beef calves has not been reported. Thus, we report here a prospective study in three herds of crossbred beef calves (n = 817) with endemic BCV. Serial blood samples for measurement of serum anti-BCV antibody titers and nasal swabs for detection of BCV and other common viral and bacterial BRD pathogens were collected from all calves or subsets of calves at predetermined times from birth through weaning. The calves were monitored for BRD and those that developed signs of respiratory disease were sampled for diagnostic testing. To discover additional risk factors that could have influenced BRD development, sequence analysis of the BCV strain(s) circulating in each herd, and the prevalence of common opportunistic bacterial pathogens in the upper respiratory tract of sick and apparently healthy cattle were also evaluated. RESULTS: Two hundred forty-eight of the 817 study calves (30.4%) were treated for BRD prior to weaning; 246 of those were from a single herd involved in two outbreaks of BRD leading to mass treatment of all calves in that group. Molecular diagnostic testing found BCV and Histophilus somni in nasal swabs taken at the time of BRD treatment. Between herd analyses revealed anti-BCV serum antibody abundance did not associate with the incidence of BRD or BCV shedding, though these measurements may have been hindered by the long periods between sample collections. Analysis of the BCV spike gene hypervariable region revealed four polymorphisms in 15 isolates from the three herds, making strain variation unlikely to account for differences in treatment rates between herds. Persistent or recurrent shedding episodes of BCV occurred in some animals treated for BRD. CONCLUSION: Co-detection of BCV and H. somni at the time of the disease outbreak suggests that these pathogens contributed to disease pathogenesis. Developing appropriate control measures for respiratory BCV infections may help decrease the incidence of pre-weaning BRD. The role of antibodies in protection must still be further defined.


Assuntos
Doenças dos Bovinos/imunologia , Infecções por Coronavirus/veterinária , Coronavirus Bovino/imunologia , Imunidade Humoral/imunologia , Animais , Anticorpos Antivirais/sangue , Bovinos , Doenças dos Bovinos/microbiologia , Coinfecção/veterinária , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/microbiologia , Coronavirus Bovino/genética , Pasteurellaceae/fisiologia , Polimorfismo Genético , Eliminação de Partículas Virais
11.
Semin Diagn Pathol ; 36(3): 152-159, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31054790

RESUMO

Lower respiratory infections remain one of the top global causes of death and the emergence of new diseases continues to be a concern. In the first two decades of the 21st century, we have born witness to the emergence of newly recognized coronaviruses that have rapidly spread around the globe, including severe acute respiratory syndrome virus (SARS) and Middle Eastern respiratory syndrome virus (MERS). We have also experienced the emergence of a novel H1N1 pandemic influenza strain in 2009 that caused substantial morbidity and mortality around the world and has transitioned into a seasonal strain. Although we perhaps most frequently think of viruses when discussing emerging respiratory infections, bacteria have not been left out of the mix, as we have witnessed an increase in the number of infections from Legionella spp. since the organisms' initial discovery in 1976. Here, we explore the basic epidemiology, clinical presentation, histopathology, and clinical laboratory diagnosis of these four pathogens and emphasize themes in humans' evolving relationship with our natural environment that have contributed to the infectious burden. Histology alone is rarely diagnostic for these infections, but has been crucial to bettering our understanding of these diseases. Together, we rely on the diagnostic acumen of pathologists to identify the clinicopathologic features that raise the suspicion of these diseases and lead to the early control of the spread in our populations.


Assuntos
Infecções por Coronavirus/patologia , Influenza Humana/patologia , Legionelose/patologia , Infecções Respiratórias/patologia , Síndrome Respiratória Aguda Grave/patologia , Infecções por Coronavirus/diagnóstico , Humanos , Influenza Humana/diagnóstico , Legionelose/diagnóstico , Infecções Respiratórias/diagnóstico , Síndrome Respiratória Aguda Grave/diagnóstico
12.
Int J Mol Sci ; 20(9)2019 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-31064083

RESUMO

Avian infectious bronchitis virus (IBV) causes considerable economic losses in the poultry industry worldwide, including Taiwan. IBV is among the most important pathogens in chickens, and it spreads rapidly among flocks. In addition to dozens of known serotypes, new viral variants have emerged due to the viral evolution and antigenic variation in IBVs. Therefore, the development of a sensitive, specific, and easily performed assay is crucial for the rapid detection and surveillance of IBV infections. A rapid and simple immunochromatographic strip (ICS) was developed in this study by employing monoclonal antibodies against spike and nucleocapsid proteins of IBV as the tracer and the capture antibody. The ICS showed high specificity in detecting IBV antigens, including several IBV genotypes and novel variants, as opposed to three other common avian respiratory viruses. The detection limit of the strip reached 104.4 50% embryo-infective dose. Moreover, in the experimental chicken model, the strip test demonstrated consistency in detecting IBV with RT-PCR gene detection. Taken together, this antigen detection strip has the potential to serve as an on-farm rapid test for IBV; therefore, it may facilitate surveillance and control of the disease.


Assuntos
Infecções por Coronavirus/diagnóstico , Vírus da Bronquite Infecciosa/imunologia , Técnicas de Diagnóstico Molecular/veterinária , Doenças das Aves Domésticas/diagnóstico , Animais , Antígenos Virais/imunologia , Galinhas , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/virologia , Feminino , Imunoensaio/métodos , Imunoensaio/normas , Imunoensaio/veterinária , Camundongos , Camundongos Endogâmicos BALB C , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Diagnóstico Molecular/normas , Doenças das Aves Domésticas/virologia , Fitas Reagentes/normas
13.
Semin Diagn Pathol ; 36(3): 197-202, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31072653

RESUMO

Infections are common during travel, and frontline physicians frequently must evaluate sick returned travelers. Sick travelers can be clinically challenging due to the wide range of endemic diseases in different geographic regions. To guide the diagnostic and treatment plan, consideration of endemic and emerging infections in the region of travel, as well as careful review of the travelers' exposures and preventative measures are necessary. Routine laboratory tests and cultures cannot confirm many tropical infections, and pathogen directed testing is typically required. Common tropical infections that can be severe, such as malaria, dengue, and enteric fever, should always be considered in the diagnostic evaluation. Providers should also be vigilant for rare but highly pathogenic emerging infections such as Ebola virus disease and Middle East respiratory syndrome (MERS).


Assuntos
Infecções por Coronavirus/diagnóstico , Dengue/diagnóstico , Doença pelo Vírus Ebola/diagnóstico , Malária/diagnóstico , Febre Tifoide/diagnóstico , Infecções por Coronavirus/patologia , Dengue/patologia , Doença pelo Vírus Ebola/patologia , Humanos , Malária/patologia , Viagem , Medicina Tropical , Febre Tifoide/patologia
14.
BMC Vet Res ; 15(1): 116, 2019 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-30987635

RESUMO

BACKGROUND: Porcine epidemic diarrhea virus (PEDV) is a major etiological agent of porcine epidemic diarrhea around the world. Point-of-care testing in the field is lacking owing to the requirement for a simple, robust field applicable test that does not require professional laboratory equipment. The aim of this study was to establish a novel reverse transcription polymerase spiral reaction (RT-PSR) assay for the rapid detection of porcine epidemic diarrhea virus (PEDV). For the assay, a specific RT-PSR primer pair was designed against a conserved region in PEDV ORF3. RESULTS: The RT-PSR was optimized, and PEDV could be detected after a 50 min incubation at 62 °C, in addition to the 15 min required for reverse transcription. No cross-reaction with other porcine infectious viruses was observed. This new method for PEDV detection was 10 times more sensitive than the conventional reverse transcription-polymerase chain reaction (RT-PCR) assay. The positive rates for 65 clinical samples using the new RT-PSR assay and the conventional RT-PCR assay were 58.46% (38/65) and 53.84% (35/65), respectively. In the RT-PSR assay, the addition of a mixture of dyes allowed a positive reaction to be directly observed by the naked eye. CONCLUSIONS: These results indicate that this RT-PSR assay is capable of accurately detecting PEDV, and has the advantages of high specificity and sensitivity for the detection of PEDV.


Assuntos
Infecções por Coronavirus/veterinária , Vírus da Diarreia Epidêmica Suína , Doenças dos Suínos/diagnóstico , Animais , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/virologia , Vírus da Diarreia Epidêmica Suína/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Sensibilidade e Especificidade , Suínos , Doenças dos Suínos/virologia
15.
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
16.
BMC Infect Dis ; 19(1): 351, 2019 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-31029095

RESUMO

BACKGROUND: Middle East respiratory syndrome coronavirus (MERS-CoV) is considered to be responsible for a new viral epidemic and an emergent threat to global health security. This study describes the current epidemiological status of MERS-CoV in the world. METHODS: Epidemiological analysis was performed on data derived from all MERS-CoV cases recorded in the disease outbreak news on WHO website between 1.1.2017 and 17.1.2018. Demographic and clinical information as well as potential contacts and probable risk factors for mortality were extracted based on laboratory-confirmed MERS-CoV cases. RESULTS: A total of 229 MERS-CoV cases, including 70 deaths (30.5%), were recorded in the disease outbreak news on world health organization website over the study period. Based on available details in this study, the case fatality rate in both genders was 30.5% (70/229) [32.1% (55/171) for males and 25.8% (15/58) for females]. The disease occurrence was higher among men [171 cases (74.7%)] than women [58 cases (25.3%)]. Variables such as comorbidities and exposure to MERS-CoV cases were significantly associated with mortality in people affected with MERS-CoV infections, and adjusted odds ratio estimates were 2.2 (95% CI: 1.16, 7.03) and 2.3 (95% CI: 1.35, 8.20), respectively. All age groups had an equal chance of mortality. CONCLUSIONS: In today's "global village", there is probability of MERS-CoV epidemic at any time and in any place without prior notice. Thus, health systems in all countries should implement better triage systems for potentially imported cases of MERS-CoV to prevent large epidemics.


Assuntos
Infecções por Coronavirus/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/mortalidade , Estudos Transversais , Demografia , Surtos de Doenças , Feminino , Saúde Global , Humanos , Masculino , Pessoa de Meia-Idade , Coronavírus da Síndrome Respiratória do Oriente Médio/isolamento & purificação , Razão de Chances , Fatores de Risco , Organização Mundial da Saúde , Adulto Jovem
17.
Diagn Microbiol Infect Dis ; 94(4): 349-354, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30929995

RESUMO

The goal of the study was to adapt our Middle East respiratory syndrome coronavirus (MERS-CoV) lab-developed test (LDT) to 3 "Sample to Result" (S2R) systems: BD MAX (BD), ELITe InGenius (ELITechGroup), and ARIES (Luminex). The BD MAX and InGenius system allowed use of lab-developed primers and TaqMan probes, while ARIES required conversion to MultiCode primers for melting curve analysis. Each device required ≤1 day of training and assay optimization. No discordant results were noted after analysis of 32 External Quality Control (EQC) samples. On a 10-fold dilution series of a MERS-CoV-positive EQC sample, InGenius obtained the highest detection rate. Laboratory technicians rated the ARIES as the user-friendliest. It also required the least hands-on time. BD MAX had the lowest turnaround time and highest throughput. While each device had distinguishing system properties with associated (dis)advantages, the 3 S2R systems were comparable in terms of assay development and validation.


Assuntos
Coronavírus da Síndrome Respiratória do Oriente Médio/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase em Tempo Real/normas , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Primers do DNA , Coronavírus da Síndrome Respiratória do Oriente Médio/genética , RNA Viral/genética
18.
Mikrochim Acta ; 186(4): 224, 2019 03 07.
Artigo em Inglês | MEDLINE | ID: mdl-30847572

RESUMO

The Middle East respiratory syndrome corona virus (MERS-CoV) is highly pathogenic. An immunosensor for the determination of MERS-CoV is described here. It is based on a competitive assay carried out on an array of carbon electrodes (DEP) modified with gold nanoparticles. Recombinant spike protein S1 was used as a biomarker for MERS CoV. The electrode array enables multiplexed detection of different CoVs. The biosensor is based on indirect competition between free virus in the sample and immobilized MERS-CoV protein for a fixed concentration of antibody added to the sample. Voltammetric response is detected by monitoring the change in the peak current (typically acquired at a working potential of -0.05 V vs. Ag/AgCl) after addition of different concentrations of antigen against MERS-CoV. Electrochemical measurements using ferrocyanide/ferricyanide as a probe were recorded using square wave voltammetry (SWV). Good linear response between the sensor response and the concentrations from 0.001 to 100 ng.mL-1 and 0.01 to 10,000 ng.mL-1 were observed for MERS-CoV and HCoV, respectively. The assay was performed in 20 min with detection limit as low as 0.4 and 1.0 pg.mL-1 for HCoV and MERS-CoV, respectively. The method is highly selective over non-specific proteins such as Influenza A and B. The method is single-step, sensitive and accurate. It was successfully applied to spiked nasal samples. Graphical abstract An electrochemical immunoassay is described for the Middle East Respiratory Syndrome Corona Virus (MERS-CoV). The method is based on a competitive assay carried out on a carbon array electrodes (DEP) nanostructured with gold nanoparticles. The array electrodes enable the multiplexed detection of different types of Corona Virus.


Assuntos
Infecções por Coronavirus/diagnóstico , Ouro/química , Nanopartículas Metálicas/química , Coronavírus da Síndrome Respiratória do Oriente Médio/química , Anticorpos Antivirais/metabolismo , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Eletrodos , Ferrocianetos/química , Imunoensaio/métodos , Limite de Detecção , Oxirredução , Tamanho da Partícula , Sensibilidade e Especificidade , Propriedades de Superfície
19.
mSphere ; 4(2)2019 03 13.
Artigo em Inglês | MEDLINE | ID: mdl-30867325

RESUMO

This study compared the performances of three commercial transmissible gastroenteritis virus/porcine respiratory coronavirus (TGEV/PRCV) blocking enzyme-linked immunosorbent assays (ELISAs) using serum samples (n = 528) collected over a 49-day observation period from pigs inoculated with TGEV strain Purdue (n = 12), TGEV strain Miller (n = 12), PRCV (n = 12), or with virus-free culture medium (n = 12). ELISA results were evaluated both with "suspect" results interpreted as positive and then as negative. All commercial kits showed excellent diagnostic specificity (99 to 100%) when testing samples from pigs inoculated with virus-free culture medium. However, analyses revealed differences between the kits in diagnostic sensitivity (percent TGEV- or PRCV-seropositive pigs), and all kits showed significant (P < 0.05) cross-reactivity between TGEV and PRCV serum antibodies, particularly during early stages of the infections. Serologic cross-reactivity between TGEV and PRCV seemed to be TGEV strain dependent, with a higher percentage of PRCV-false-positive results for pigs inoculated with TGEV Purdue than for TGEV Miller. Moreover, the overall proportion of false positives was higher when suspect results were interpreted as positive, regardless of the ELISA kit evaluated.IMPORTANCE Current measures to prevent TGEV from entering a naive herd include quarantine and testing for TGEV-seronegative animals. However, TGEV serology is complicated due to the cross-reactivity with PRCV, which circulates subclinically in most swine herds worldwide. Conventional serological tests cannot distinguish between TGEV and PRCV antibodies; however, blocking ELISAs using antigen containing a large deletion in the amino terminus of the PRCV S protein permit differentiation of PRCV and TGEV antibodies. Several commercial TGEV/PRCV blocking ELISAs are available, but performance comparisons have not been reported in recent research. This study demonstrates that the serologic cross-reactivity between TGEV and PRCV affects the accuracy of commercial blocking ELISAs. Individual test results must be interpreted with caution, particularly in the event of suspect results. Therefore, commercial TGEV/PRCV blocking ELISAs should only be applied on a herd basis.


Assuntos
Anticorpos Antivirais/análise , Diarreia/veterinária , Ensaio de Imunoadsorção Enzimática/métodos , Coronavirus Respiratório Porcino/imunologia , Vírus da Gastroenterite Transmissível/imunologia , Animais , Antígenos Virais/imunologia , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/veterinária , Reações Cruzadas , Reações Falso-Positivas , Gastroenterite Suína Transmissível/diagnóstico , Kit de Reagentes para Diagnóstico/normas , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/veterinária , Sensibilidade e Especificidade , Suínos
20.
Appl Microbiol Biotechnol ; 103(9): 3705-3714, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30877355

RESUMO

The variant strains of porcine epidemic diarrhea virus (PEDV) severely threaten the pig industry worldwide and cause up to 100% mortality in suckling piglets. It is critically important and urgent to develop tools for detection of PEDV infection. In this study, we developed six monoclonal antibodies (mAbs) targeting N protein of PEDV and analyzed their applications on enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA), western blot assay, and flow cytometry assay. The results demonstrated that all these six mAbs were IgG1 isotype and κ chain. Among these six mAbs, 3F12 recognizes a linear epitope (VAAVKDALKSLGI) while the other five mAbs recognize different conformational epitopes formed by a specific peptide fragment or the full length of N protein. The functional analysis showed that all these six mAbs were applicable to ELISA, western blot, IFA, and flow cytometry assay. In conclusion, we developed six mAbs against PEDV-N protein to facilitate the early detection of PEDV infection using ELISA, western blot, IFA, and flow cytometry.


Assuntos
Anticorpos Monoclonais/análise , Anticorpos Antivirais/análise , Infecções por Coronavirus/veterinária , Nucleocapsídeo/análise , Vírus da Diarreia Epidêmica Suína/isolamento & purificação , Doenças dos Suínos/virologia , Animais , Anticorpos Antivirais/imunologia , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/virologia , Ensaio de Imunoadsorção Enzimática , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Imunização , Camundongos Endogâmicos BALB C , Nucleocapsídeo/imunologia , Vírus da Diarreia Epidêmica Suína/imunologia , Suínos , Doenças dos Suínos/diagnóstico
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