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1.
Int J Mol Sci ; 22(24)2021 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-34948342

RESUMO

Although blood-heart-barrier (BHB) leakage is the hallmark of congestive (cardio-pulmonary) heart failure (CHF), the primary cause of death in elderly, and during viral myocarditis resulting from the novel coronavirus variants such as the severe acute respiratory syndrome novel corona virus 2 (SARS-CoV-2) known as COVID-19, the mechanism is unclear. The goal of this project is to determine the mechanism of the BHB in CHF. Endocardial endothelium (EE) is the BHB against leakage of blood from endocardium to the interstitium; however, this BHB is broken during CHF. Previous studies from our laboratory, and others have shown a robust activation of matrix metalloproteinase-9 (MMP-9) during CHF. MMP-9 degrades the connexins leading to EE dysfunction. We demonstrated juxtacrine coupling of EE with myocyte and mitochondria (Mito) but how it works still remains at large. To test whether activation of MMP-9 causes EE barrier dysfunction, we hypothesized that if that were the case then treatment with hydroxychloroquine (HCQ) could, in fact, inhibit MMP-9, and thus preserve the EE barrier/juxtacrine signaling, and synchronous endothelial-myocyte coupling. To determine this, CHF was created by aorta-vena cava fistula (AVF) employing the mouse as a model system. The sham, and AVF mice were treated with HCQ. Cardiac hypertrophy, tissue remodeling-induced mitochondrial-myocyte, and endothelial-myocyte contractions were measured. Microvascular leakage was measured using FITC-albumin conjugate. The cardiac function was measured by echocardiography (Echo). Results suggest that MMP-9 activation, endocardial endothelial leakage, endothelial-myocyte (E-M) uncoupling, dyssynchronous mitochondrial fusion-fission (Mfn2/Drp1 ratio), and mito-myocyte uncoupling in the AVF heart failure were found to be rampant; however, treatment with HCQ successfully mitigated some of the deleterious cardiac alterations during CHF. The findings have direct relevance to the gamut of cardiac manifestations, and the resultant phenotypes arising from the ongoing complications of COVID-19 in human subjects.


Assuntos
COVID-19/complicações , Insuficiência Cardíaca/metabolismo , Coração/virologia , Animais , Sangue/virologia , Fenômenos Fisiológicos Sanguíneos/imunologia , COVID-19/fisiopatologia , Cardiomegalia/metabolismo , Doenças Cardiovasculares/metabolismo , Fenômenos Fisiológicos Cardiovasculares/imunologia , Modelos Animais de Doenças , Endotélio/metabolismo , Coração/fisiopatologia , Insuficiência Cardíaca/virologia , Hidroxicloroquina/farmacologia , Masculino , Metaloproteinase 9 da Matriz/efeitos dos fármacos , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Células Musculares/metabolismo , Miocárdio/metabolismo , SARS-CoV-2/metabolismo , SARS-CoV-2/patogenicidade , Remodelação Ventricular/fisiologia
2.
Prev Vet Med ; 193: 105397, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34147958

RESUMO

The aim of this study was to compare the detection of porcine reproductive and respiratory syndrome virus (PRRSV) in due-to-wean litters in commercial swine breeding herds using family oral fluids (FOF) vs. individual piglet serum samples. FOF and piglet serum samples were collected in 199 due-to-wean litters on six farms containing 2177 piglets. All samples were individually tested for PRRSV RNA by RT-rtPCR. A litter was considered PRRSV-positive when PRRSV RNA was detected in ≥ 1 piglet serum sample or the FOF sample. Mixed effect logistic regression with farm as a random effect was used 1) to evaluate the probability of obtaining a PRRSV RNA positive FOF as a function of the proportion of viremic piglets in a litter and 2) the effect of litter size and parity on the probability that a litter would test PRRSV RNA positive in FOF. A Bayesian prevalence estimation under misclassification (BayesPEM) analysis was used to calculate the PRRSV prevalence and 95 % credible interval given the condition that all samples (FOF and serum) tested negative. In total, 34 of 199 litters (17.1 %) contained ≥ 1 viremic piglet(s), and 28 of 199 litters (14.1 %) were FOF positive. When all piglet serum samples within a litter tested negative, 1 of 165 FOF (0.6 %) tested PRRSV RNA positive. The probability of a PCR-positive FOF sample from litters with 10 %, 20 %, 30 %, 40 %, and 50 % within-litter PRRSV prevalence was 3.5 %, 35.1 %, 88.8 %, 99.2 %, and >99.9 %, respectively. The odds of a PCR-positive FOF in a first parity litter were 3.36 times (95 % CI: 2.10-5.38) that of a parity ≥ 2 litter. The odds of a positive FOF result in a litter with ≤ 11 piglets were 9.90 times (95 % CI: 4.62-21.22) that of a litter with > 11 piglets. FOF was shown to be an efficacious sample type for PRRSV detection in farrowing rooms. A risk-based approach for litter selection combined with FOF collection can be used to improve on-farm PRRSV detection with a limited sample size, compared to sampling multiple individual pigs. Finally, the BayesPEM analysis showed that PRRSV may still be present in breeding herds when all samples (serum and FOF) test PRRSV RNA negative, i.e., negative surveillance results should be interpreted with caution.


Assuntos
Síndrome Respiratória e Reprodutiva Suína , Vírus da Síndrome Respiratória e Reprodutiva Suína , Animais , Teorema de Bayes , Sangue/virologia , Feminino , Tamanho da Ninhada de Vivíparos , Síndrome Respiratória e Reprodutiva Suína/diagnóstico , Síndrome Respiratória e Reprodutiva Suína/epidemiologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/isolamento & purificação , Gravidez , Saliva/virologia , Suínos , Desmame
3.
Vet Res ; 52(1): 91, 2021 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-34158102

RESUMO

Understanding the mechanisms of transmission of infectious laryngotracheitis virus (ILTV) is critical to proper control as both vaccine and wild-type strains circulate within chicken flocks with potential adverse consequences. The relative efficiency of transmission by direct contact between chickens and airborne transmission has not been investigated. Furthermore, relatively high levels of ILTV DNA have been detected in poultry dust and blood but the infectivity of these is unknown. In this study, comparison of in-contact and airborne transmission of two vaccine and one field strain of ILTV revealed that all transmitted to 100% of in-contact birds by 6 days post-exposure (dpe). Airborne transmission without contact resulted in 100% transmission by 14 and 17 dpe for the wild-type and Serva vaccine virus but only 27% transmission by 21 dpe for the A20 vaccine virus. The infectivity of dust or extracts of dust and blood or plasma from infected chickens at various stages of infection was assessed by inoculation into susceptible chickens. There was no transmission by any of these materials. In conclusion, direct contact facilitated efficient ILTV transmission but the virus was unable to be transmitted by dust from infected chickens suggestive of a limited role in the epidemiology of ILTV.


Assuntos
Poeira , Infecções por Herpesviridae/veterinária , Herpesvirus Galináceo 1/fisiologia , Vacinas contra Herpesvirus/efeitos adversos , Doenças das Aves Domésticas/transmissão , Animais , Sangue/virologia , Galinhas , Infecções por Herpesviridae/transmissão , Infecções por Herpesviridae/virologia , Abrigo para Animais , Plasma/virologia , Doenças das Aves Domésticas/virologia , Replicação Viral
4.
BMC Infect Dis ; 21(1): 501, 2021 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-34051756

RESUMO

BACKGROUND: Tick-borne pathogens other than Borrelia burgdorferi sensu lato - the causative agent of Lyme borreliosis - are common in Ixodes ricinus ticks. How often these pathogens cause human disease is unknown. In addition, diagnostic tools to identify such diseases are lacking or reserved to research laboratories. To elucidate their prevalence and disease burden, the study 'Ticking on Pandora's Box' has been initiated, a collaborative effort between Amsterdam University Medical Center and the National Institute for Public Health and the Environment. METHODS: The study investigates how often the tick-borne pathogens Anaplasma phagocytophilum, Babesia species, Borrelia miyamotoi, Neoehrlichia mikurensis, spotted fever group Rickettsia species and/or tick-borne encephalitis virus cause an acute febrile illness after tick-bite. We aim to determine the impact and severity of these tick-borne diseases in the Netherlands by measuring their prevalence and describing their clinical picture and course of disease. The study is designed as a prospective case-control study. We aim to include 150 cases - individuals clinically suspected of a tick-borne disease - and 3 matched healthy control groups of 200 persons each. The controls consist respectively of a group of individuals with either a tick-bite without complaints, the general population and of healthy blood donors. During a one-year follow-up we will acquire blood, urine and skin biopsy samples and ticks at baseline, 4 and 12 weeks. Additionally, participants answer modified versions of validated questionnaires to assess self-reported symptoms, among which the SF-36, on a 3 monthly basis. DISCUSSION: This article describes the background and design of the study protocol of 'Ticking on Pandora's Box'. With our study we hope to provide insight into the prevalence, clinical presentation and disease burden of the tick-borne diseases anaplasmosis, babesiosis, B. miyamotoi disease, neoehrlichiosis, rickettsiosis and tick-borne encephalitis and to assist in test development as well as provide recommendations for national guidelines. TRIAL REGISTRATION: NL9258 (retrospectively registered at Netherlands Trial Register, trialregister.nl in in February 2021).


Assuntos
Ixodes/microbiologia , Doenças Transmitidas por Carrapatos/epidemiologia , Doenças Transmitidas por Carrapatos/microbiologia , Adulto , Animais , Sangue/microbiologia , Sangue/virologia , Estudos de Casos e Controles , DNA Bacteriano , Febre/epidemiologia , Febre/microbiologia , Febre/virologia , Seguimentos , Humanos , Pessoa de Meia-Idade , Países Baixos/epidemiologia , Prevalência , Estudos Prospectivos , Reação em Cadeia da Polimerase em Tempo Real , Índice de Gravidade de Doença , Pele/microbiologia , Pele/virologia , Inquéritos e Questionários , Picadas de Carrapatos/epidemiologia , Picadas de Carrapatos/microbiologia , Picadas de Carrapatos/virologia , Urina/microbiologia , Urina/virologia
5.
Parasit Vectors ; 14(1): 194, 2021 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-33832527

RESUMO

BACKGROUND: Infectious blood meal experiments have been frequently performed with different virus-vector combinations to assess the transmission potential of arthropod-borne (arbo)viruses. A wide variety of host blood sources have been used to deliver arboviruses to their arthropod vectors in laboratory studies. The type of blood used during vector competence experiments does not always reflect the blood from the viremic vertebrate hosts in the field, but little is known about the effect of blood source on the experimental outcome of vector competence studies. Here we investigated the effect of avian versus human blood on the infection and transmission rates of the zoonotic Usutu virus (USUV) in its primary mosquito vector Culex pipiens. METHODS: Cx. pipiens biotypes (pipiens and molestus) were orally infected with USUV through infectious blood meals containing either chicken or human whole blood. The USUV infection and transmission rates were determined by checking mosquito bodies and saliva for USUV presence after 14 days of incubation at 28 °C. In addition, viral titers were determined for USUV-positive mosquito bodies and saliva. RESULTS: Human and chicken blood lead to similar USUV transmission rates for Cx. pipiens biotype pipiens (18% and 15%, respectively), while human blood moderately but not significantly increased the transmission rate (30%) compared to chicken blood (17%) for biotype molestus. USUV infection rates with human blood were consistently higher in both Cx. pipiens biotypes compared to chicken blood. In virus-positive mosquitoes, USUV body and saliva titers did not differ between mosquitoes taking either human or chicken blood. Importantly, biotype molestus had much lower USUV saliva titers compared to biotype pipiens, regardless of which blood was offered. CONCLUSIONS: Infection of mosquitoes with human blood led to higher USUV infection rates as compared to chicken blood. However, the blood source had no effect on the vector competence for USUV. Interestingly, biotype molestus is less likely to transmit USUV compared to biotype pipiens due to very low virus titers in the saliva.


Assuntos
Culex/fisiologia , Infecções por Flavivirus/veterinária , Infecções por Flavivirus/virologia , Flavivirus/fisiologia , Mosquitos Vetores/fisiologia , Doenças das Aves Domésticas/virologia , Animais , Sangue/virologia , Galinhas/virologia , Culex/virologia , Comportamento Alimentar , Flavivirus/genética , Flavivirus/isolamento & purificação , Infecções por Flavivirus/sangue , Infecções por Flavivirus/transmissão , Humanos , Mosquitos Vetores/virologia , Doenças das Aves Domésticas/sangue , Doenças das Aves Domésticas/transmissão , Zoonoses Virais/transmissão , Zoonoses Virais/virologia
6.
J Med Virol ; 93(8): 5134-5140, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-33837954

RESUMO

Blood product transfusion can transmit viral pathogens. Pathogen reduction methods for blood products have been developed but, so far, are not available for whole blood. We evaluated if vitamin K5 (VK5) and ultraviolet A (UVA) irradiation could be used for virus inactivation in plasma and whole blood. Undiluted human plasma and whole blood diluted to 20% were spiked with high levels of vaccinia or Zika viruses. Infectious titers were measured by standard TCID50 assay before and after VK5/UVA treatments. Up to 3.6 log of vaccinia and 3.2 log of Zika were reduced in plasma by the combination of 500 µM VK5 and 3 J/cm2 UVA, and 3.1 log of vaccinia and 2.9 log of Zika were reduced in diluted human blood (20%) by the combination of 500 µM VK5 and 70 J/cm2 UVA. At end of whole blood treatment, hemolysis increased from 0.18% to 0.41% but remained below 1% hemolysis, which is acceptable to the Food and Drug Administration for red cell transfusion products. No significant alteration of biochemical parameters of red blood cells occurred with treatment. Our results provide proof of the concept that a viral pathogen reduction method based on VK5/UVA may be developed for whole blood.


Assuntos
Segurança do Sangue/métodos , Sangue/virologia , Fármacos Fotossensibilizantes/farmacologia , Inativação de Vírus/efeitos dos fármacos , Vitamina K 3/análogos & derivados , Sangue/efeitos dos fármacos , Segurança do Sangue/normas , Transfusão de Sangue/normas , Hemólise/efeitos dos fármacos , Humanos , Fármacos Fotossensibilizantes/efeitos da radiação , Raios Ultravioleta , Vírus Vaccinia/efeitos dos fármacos , Viroses/prevenção & controle , Vitamina K 3/farmacologia , Vitamina K 3/efeitos da radiação , Zika virus/efeitos dos fármacos
7.
PLoS Negl Trop Dis ; 15(4): e0009336, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33872309

RESUMO

BACKGROUND: Serological diagnosis of Zika virus (ZIKV) infection is challenging because of the antibody cross-reactivity among flaviviruses. At the same time, the role of Nucleic Acid Testing (NAT) is limited by the low proportion of symptomatic infections and the low average viral load. Here, we compared the diagnostic performance of commercially available IgM, IgAM, and IgG ELISAs in sequential samples during the ZIKV and chikungunya (CHIKV) epidemics and co-circulation of dengue virus (DENV) in Brazil and Venezuela. METHODOLOGY/PRINCIPAL FINDINGS: Acute (day of illness 1-5) and follow-up (day of illness ≥ 6) blood samples were collected from nine hundred and seven symptomatic patients enrolled in a prospective multicenter study between June 2012 and August 2016. Acute samples were tested by RT-PCR for ZIKV, DENV, and CHIKV. Acute and follow-up samples were tested for IgM, IgAM, and IgG antibodies to ZIKV using commercially available ELISAs. Among follow-up samples with a RT-PCR confirmed ZIKV infection, anti-ZIKV IgAM sensitivity was 93.5% (43/46), while IgM and IgG exhibited sensitivities of 30.3% (10/33) and 72% (18/25), respectively. An additional 24% (26/109) of ZIKV infections were detected via IgAM seroconversion in ZIKV/DENV/CHIKV RT-PCR negative patients. The specificity of anti-ZIKV IgM was estimated at 93% and that of IgAM at 85%. CONCLUSIONS/SIGNIFICANCE: Our findings exemplify the challenges of the assessment of test performance for ZIKV serological tests in the real-world setting, during co-circulation of DENV, ZIKV, and CHIKV. However, we can also demonstrate that the IgAM immunoassay exhibits superior sensitivity to detect ZIKV RT-PCR confirmed infections compared to IgG and IgM immunoassays. The IgAM assay also proves to be promising for detection of anti-ZIKV seroconversions in sequential samples, both in ZIKV PCR-positive as well as PCR-negative patients, making this a candidate assay for serological monitoring of pregnant women in future ZIKV outbreaks.


Assuntos
Febre de Chikungunya/diagnóstico , Dengue/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Testes Sorológicos/métodos , Infecção por Zika virus/diagnóstico , Adolescente , Adulto , Anticorpos Antivirais/sangue , Sangue/virologia , Brasil , Criança , Diagnóstico Diferencial , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , Estudos Prospectivos , RNA Viral/sangue , Reação em Cadeia da Polimerase em Tempo Real , Venezuela , Adulto Jovem
8.
Sci Rep ; 11(1): 6921, 2021 03 25.
Artigo em Inglês | MEDLINE | ID: mdl-33767340

RESUMO

Human blood metagenomics has revealed the presence of different types of viruses in apparently healthy subjects. By far, anelloviruses constitute the viral family that is more frequently found in human blood, although amplification biases and contaminations pose a major challenge in this field. To investigate this further, we subjected pooled plasma samples from 120 healthy donors in Spain to high-speed centrifugation, RNA and DNA extraction, random amplification, and massive parallel sequencing. Our results confirm the extensive presence of anelloviruses in such samples, which represented nearly 97% of the total viral sequence reads obtained. We assembled 114 different viral genomes belonging to this family, revealing remarkable diversity. Phylogenetic analysis of ORF1 suggested 28 potentially novel anellovirus species, 24 of which were validated by Sanger sequencing to discard artifacts. These findings underscore the importance of implementing more efficient purification procedures that enrich the viral fraction as an essential step in virome studies and question the suggested pathological role of anelloviruses.


Assuntos
Anelloviridae/isolamento & purificação , Sangue/virologia , Viroma , Voluntários Saudáveis , Humanos , Metagenômica
9.
PLoS Negl Trop Dis ; 15(2): e0009173, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33600413

RESUMO

BACKGROUND: As an invasive mosquito species in the United States, Aedes albopictus is a potential vector of arboviruses including dengue, chikungunya, and Zika, and may also be involved in occasional transmission of other arboviruses such as West Nile, Saint Louis encephalitis, eastern equine encephalitis, and La Crosse viruses. Aedes albopictus feeds on a wide variety of vertebrate hosts, wild and domestic, as well as humans. METHODOLOGY/PRINCIPAL FINDINGS: In order to investigate blood feeding patterns of Ae. albopictus, engorged specimens were collected from a variety of habitat types using the Centers for Disease Control and Prevention light traps, Biogents Sentinel 2 traps, and modified Reiter gravid traps in southeast Virginia. Sources of blood meals were determined by the analysis of mitochondrial cytochrome b gene sequences amplified in PCR assays. Our aims were to quantify degrees of Ae. albopictus interactions with vertebrate hosts as sources of blood meals, investigate arboviral infection status, assess the influence of key socioecological conditions on spatial variability in blood feeding, and investigate temporal differences in blood feeding by season. Analysis of 961 engorged specimens of Ae. albopictus sampled between 2017-2019 indicated that 96%, 4%, and less than 1% obtained blood meals from mammalian, reptilian, and avian hosts, respectively. Domestic cats were the most frequently identified (50.5%) hosts followed by Virginia opossums (17.1%), white-tailed deer (12.2%), and humans (7.3%), together representing 87.1% of all identified blood hosts. We found spatial patterns in blood feeding linked to socioecological conditions and seasonal shifts in Ae. albopictus blood feeding with implications for understanding human biting and disease risk. In Suffolk Virginia in areas of lower human development, the likelihood of human blood feeding increased as median household income increased and human blood feeding was more likely early in the season (May-June) compared to later (July-October). Screening of the head and thorax of engorged Ae. albopictus mosquitoes by cell culture and RT-PCR resulted in a single isolate of Potosi virus. CONCLUSION AND SIGNIFICANCE: Understanding mosquito-host interactions in nature is vital for evaluating vectorial capacity of mosquitoes. These interactions with competent reservoir hosts support transmission, maintenance, and amplification of zoonotic agents of human diseases. Results of our study in conjunction with abundance in urban/suburban settings, virus isolation from field-collected mosquitoes, and vector competence of Ae. albopictus, highlight the potential involvement of this species in the transmission of a number of arboviruses such as dengue, chikungunya, and Zika to humans. Limited interaction with avian hosts suggests that Ae. albopictus is unlikely to serve as a bridge vector of arboviruses such as West Nile and eastern equine encephalitis in the study region, but that possibility cannot be entirely ruled out.


Assuntos
Aedes/fisiologia , Aedes/virologia , Arbovírus/isolamento & purificação , Comportamento Alimentar , Animais , Infecções por Arbovirus , Aves , Sangue/virologia , Humanos , Mamíferos , Mosquitos Vetores/fisiologia , Mosquitos Vetores/virologia , Tartarugas , Virginia
11.
Parasit Vectors ; 14(1): 76, 2021 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-33482887

RESUMO

BACKGROUND: On 11 March 2020, the World Health Organisation (WHO) declared the coronavirus disease 2019 (COVID-19) outbreak to be a pandemic. As the mosquito season progressed, the understandable concern that mosquitoes could transmit the virus began to increase among the general public and public health organisations. We have investigated the vector competence of Culex pipiens and Aedes albopictus, the two most common species of vector mosquitoes in Europe, for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Due to the very unusual feeding behaviour of Ae. albopictus, we also evaluated the role of this mosquito in a potential mechanical transmission of the virus. METHODS: For the vector competence study, mosquitoes were allowed to take several infectious blood meals. The mosquitoes were then collected and analysed at 0, 3, 7 and 10 days post-feeding. For the mechanical transmission test, Ae. albopictus females were allowed to feed for a short time on a feeder containing infectious blood and then on a feeder containing virus-free blood. Both mosquitoes and blood were tested for viral presence. RESULTS: Culex pipiens and Ae. albopictus were found not be competent vectors for SARS-CoV-2, and Ae. albopictus was unable to mechanically transmit the virus. CONCLUSIONS: This is the first study to show that the most common species of vector mosquitoes in Europe do not transmit SARS-CoV-2 and that Ae. albopictus is unable to mechanically transmit the virus from a positive host to a healthy host through host-feeding.


Assuntos
Aedes/virologia , COVID-19/transmissão , Culex/virologia , Mosquitos Vetores/virologia , SARS-CoV-2/fisiologia , Animais , Sangue/virologia , Europa (Continente) , Feminino , RNA Viral/análise , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , SARS-CoV-2/genética , Ovinos/sangue
12.
J Transl Med ; 19(1): 30, 2021 01 07.
Artigo em Inglês | MEDLINE | ID: mdl-33413461

RESUMO

BACKGROUND: COVID-19 has caused a global pandemic and the death toll is increasing. However, there is no definitive information regarding the type of clinical specimens that is the best for SARS-CoV-2 detection, the antibody levels in patients with different duration of disease, and the relationship between antibody level and viral load. METHODS: Nasopharyngeal swabs, anal swabs, saliva, blood, and urine specimens were collected from patients with a course of disease ranging from 7 to 69 days. Viral load in different specimen types was measured using droplet digital PCR (ddPCR). Meanwhile, anti-nucleocapsid protein (anti-N) IgM and IgG antibodies and anti-spike protein receptor-binding domain (anti-S-RBD) IgG antibody in all serum samples were tested using ELISA. RESULTS: The positive detection rate in nasopharyngeal swab was the highest (54.05%), followed by anal swab (24.32%), and the positive detection rate in saliva, blood, and urine was 16.22%, 10.81%, and 5.41%, respectively. However, some patients with negative nasopharyngeal swabs had other specimens tested positive. There was no significant correlation between antibody level and days after symptoms onset or viral load. CONCLUSIONS: Other specimens could be positive in patients with negative nasopharyngeal swabs, suggesting that for patients in the recovery period, specimens other than nasopharyngeal swabs should also be tested to avoid false negative results, and anal swabs are recommended. The antibody level had no correlation with days after symptoms onset or the viral load of nasopharyngeal swabs, suggesting that the antibody level may also be affected by other factors.


Assuntos
Anticorpos Antivirais/sangue , COVID-19/imunologia , COVID-19/virologia , SARS-CoV-2/imunologia , SARS-CoV-2/isolamento & purificação , Carga Viral , Adulto , Idoso , Idoso de 80 Anos ou mais , Canal Anal/virologia , Sangue/virologia , COVID-19/epidemiologia , Teste Sorológico para COVID-19 , Teste para COVID-19 , China/epidemiologia , Reações Falso-Negativas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Nasofaringe/virologia , Pandemias , Saliva/virologia , Manejo de Espécimes , Fatores de Tempo , Urina/virologia
13.
Mem Inst Oswaldo Cruz ; 115: e200339, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33503145

RESUMO

We evaluated sweat, blood and urine specimens obtained from an ongoing cohort study in Brazil. Samples were collected at pre-established intervals after the initial rash presentation and tested for Zika virus (ZIKV) RNA presence by real-time reverse transcriptase polymerase chain reaction (rRT-PCR). From 254 participants with confirmed infection, ZIKV RNA was detected in the sweat of 46 individuals (18.1%). Sweat presented a median cycle threshold (Ct) of 34.74 [interquartile range (IQR) 33.44-36.04], comparable to plasma (Ct 35.96 - IQR 33.29-36.69) and higher than urine (Ct 30.78 - IQR 28.72-33.22). Concomitant detection with other specimens was observed in 33 (72%) of 46 participants who had a positive result in sweat. These findings represent an unusual and not yet investigated virus shedding through eccrine glands.


Assuntos
RNA Viral/genética , Suor/virologia , Infecção por Zika virus/diagnóstico , Zika virus/isolamento & purificação , Adulto , Sangue/virologia , Brasil/epidemiologia , Estudos de Coortes , Feminino , Humanos , Masculino , RNA Viral/classificação , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Urina/virologia , Zika virus/genética , Infecção por Zika virus/epidemiologia
14.
Vox Sang ; 116(1): 3-12, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32986873

RESUMO

Blood transfusion remains a routine life-saving medical procedure that helps replace blood lost due to surgery, injury or disease. The quality of transfused blood is crucial in this process as blood donors must be free of transfusion-transmissible infections and donated blood should be compatible to that of the recipient. The quality of donated blood could be affected by the quality of in vitro diagnostic medical devices (IVDs) used in the screening process. Consequently, the need for high-quality, safe and well-performing IVDs for use in transfusion medicine arises, accompanied by the need for tight regulations in this domain. In the European Union, the new IVD Regulation will replace the existing IVD Directive within a five-year transitional period. Manufacturers of IVDs are expected to fully comply with the new Regulation by 26 May 2022. In this review, we address the major differences relating to marketing authorization and testing between this new Regulation and its predecessor. We further present the main elements of the prequalification assessment introduced by the WHO for IVDs, including disease-specific IVDs for blood screening laboratories.


Assuntos
Transfusão de Sangue/métodos , Organização Mundial da Saúde , Sangue/microbiologia , Sangue/virologia , Análise Química do Sangue , Transfusão de Sangue/legislação & jurisprudência , Técnicas de Laboratório Clínico , Humanos , Técnicas In Vitro
15.
Sci Rep ; 10(1): 20425, 2020 11 24.
Artigo em Inglês | MEDLINE | ID: mdl-33235273

RESUMO

Using mass cytometry, we investigated the expression of 28 markers on CD8+ and CD4+ T cells from HIV-1 infected patients with a variable size of HIV-1 reservoir defined as high (HR) and low (LR) reservoir; we aimed at identifying phenotypic associations of T cells with size of HIV-1 reservoir. We showed that the frequency of CD45+ CD8+ and CD4+ T cells was directly proportional to the size of HIV-1 reservoir; HR patients had a significantly larger frequency of blood CD45high T cells and higher CD45 expression on both CD8+ and CD4+ T cells. CD45 is a receptor-type protein tyrosine phosphatase essential in TCR signaling. Functional and phenotypical analysis of CD45high cells revealed that they express activation and proliferation markers (CD38 + HLA-DR + and Ki-67) and produce cytokines upon in vitro activation. CD45high T cells also expressed high levels of immune check-point PD-1. Our results link CD45 expression on T cells to HIV-1 reservoir; PD-1 expression on CD45high T cells may contribute to their exhaustion.


Assuntos
Sangue/virologia , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Infecções por HIV/virologia , HIV-1/genética , Antígenos Comuns de Leucócito/metabolismo , Citocinas/metabolismo , Feminino , Citometria de Fluxo , Infecções por HIV/sangue , Infecções por HIV/imunologia , Humanos , Masculino , Receptor de Morte Celular Programada 1/metabolismo , RNA Viral/genética , Regulação para Cima , Carga Viral
16.
Sci Rep ; 10(1): 19847, 2020 11 16.
Artigo em Inglês | MEDLINE | ID: mdl-33199784

RESUMO

Swine influenza is one of the important zoonotic diseases of pigs. We conducted a longitudinal survey of swine influenza A viruses (S-IAV) circulating in a pig farm with history of endemic S-IAV infection from 2017 to 2018. The samples were collected from 436 pigs including nasal swab samples (n = 436) and blood samples (n = 436). Our result showed that 18.81% (82/436) were positive for influenza A virus and subsequently 57 S-IAV could be isolated. Then 24 out of 57 S-IAVs were selected for whole genome sequencing and could be subtyped as S-IAV-H1N1 (n = 18) and S-IAV-H3N2 (n = 6). Of 24 S-IAVs, we observed 3 genotypes of S-IAVs including rH1N1 (pdm + 1), rH1N1 (pdm + 2), and rH3N2 (pdm + 2). Since all genotypes of S-IAVs in this study contained internal genes from pdmH1N1-2009, it could be speculated that pdmH1N1-2009 was introduced in a pig farm and then multiple reassorted with endemic S-IAVs to generate diversify S-IAV genotypes. Our study supported and added the evidences that pdmH1N1-2009 and it reassortant have predominately persisted in pig population in Thailand. Thus, monitoring of S-IAVs in pigs, farm workers and veterinarians in pig farms is important and should be routinely conducted.


Assuntos
Vírus da Influenza A Subtipo H1N1/classificação , Infecções por Orthomyxoviridae/epidemiologia , RNA Viral/genética , Vírus Reordenados/classificação , Sequenciamento Completo do Genoma/métodos , Animais , Animais Domésticos/virologia , Sangue/virologia , Técnicas de Genotipagem , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Estudos Longitudinais , Nariz/virologia , Infecções por Orthomyxoviridae/virologia , Filogenia , Vírus Reordenados/genética , Vírus Reordenados/isolamento & purificação , Suínos , Tailândia/epidemiologia
17.
Arch Virol ; 165(12): 2847-2856, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33034764

RESUMO

Here, we investigated the fecal, oral, blood, and skin virome of 10 laboratory rabbits using a viral metagenomic method. In the oral samples, we detected a novel polyomavirus (RabPyV), and phylogenetic analysis based on the large T antigen, VP1 and VP2 regions indicated that the novel strain might have undergone a recombination event. Recombination analysis based on related genomes confirmed that RabPyV is a multiple recombinant between rodent-like and avian-like polyomaviruses. In fecal samples, three partial or complete genome sequences of viruses belonging to the families Picobirnaviridae, Parvoviridae, Microviridae and Coronaviridae were characterized, and phylogenetic trees were constructed based on the predicted amino acid sequences of viral proteins. This study increases the amount of genetic information on viruses present in laboratory rabbits.


Assuntos
Metagenoma , Polyomavirus/isolamento & purificação , Coelhos/virologia , Proteínas Virais/genética , Vírus/classificação , Animais , Animais de Laboratório/virologia , Sangue/virologia , Fezes/virologia , Genoma Viral , Boca/virologia , Filogenia , Pele/virologia , Vírus/isolamento & purificação , Sequenciamento Completo do Genoma
18.
Viruses ; 12(10)2020 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-33019736

RESUMO

Europe is currently experiencing a long-lasting African swine fever (ASF) epidemic, both in domestic pigs and wild boar. There is great concern that carcasses of infected wild boar may act as long-term virus reservoirs in the environment. We evaluated the tenacity of ASF virus (ASFV) in tissues and body fluids from experimentally infected domestic pigs and wild boar, which were stored on different matrices and at different temperatures. Samples were analysed at regular intervals for viral genome and infectious virus. ASFV was most stable in spleen or muscles stored at -20 °C and in blood stored at 4 °C. In bones stored at -20 °C, infectious virus was detected for up to three months, and at 4 °C for up to one month, while at room temperature (RT), no infectious virus could be recovered after one week. Skin stored at -20 °C, 4 °C and RT remained infectious for up to three, six and three months, respectively. In urine and faeces, no infectious virus was recovered after one week, irrespective of the matrix. In conclusion, tissues and organs from decomposing carcasses that persist in the environment for a long time can be a source of infection for several months, especially at low temperatures.


Assuntos
Vírus da Febre Suína Africana/isolamento & purificação , Vírus da Febre Suína Africana/fisiologia , Febre Suína Africana/epidemiologia , Sus scrofa/virologia , Vírus da Febre Suína Africana/genética , Animais , Sangue/virologia , Medula Óssea/virologia , Estônia , Fezes/virologia , Genoma Viral , Cinética , Músculos/virologia , Fatores de Risco , Pele/virologia , Baço/virologia , Suínos , Temperatura , Urina/virologia
19.
Viruses ; 12(9)2020 09 04.
Artigo em Inglês | MEDLINE | ID: mdl-32899621

RESUMO

Laboratory diagnosis of human T-lymphotropic viruses (HTLV) 1 and 2 infection is performed by serological screening and further confirmation with serological or molecular assays. Thus, we developed a loop-mediated isothermal nucleic acid amplification (LAMP) assay for the detection of HTLV-1/2 in blood samples. The sensitivity and accuracy of HTLV-1/2 LAMP were defined with DNA samples from individuals infected with HTLV-1 (n = 125), HTLV-2 (n = 19), and coinfected with HIV (n = 82), and compared with real-time polymerase chain reaction (qPCR) and PCR-restriction fragment length polymorphism (RFLP). The overall accuracy of HTLV-1/2 LAMP (95% CI 74.8-85.5%) was slightly superior to qPCR (95% CI 69.5-81.1%) and similar to PCR-RFLP (95% CI 79.5-89.3%). The sensitivity of LAMP was greater for HTLV-1 (95% CI 83.2-93.4%) than for HTLV-2 (95% CI 43.2-70.8%). This was also observed in qPCR and PCR-RFLP, which was associated with the commonly lower HTLV-2 proviral load. All molecular assays tested showed better results with samples from HTLV-1/2 mono-infected individuals compared with HIV-coinfected patients, who present lower CD4 T-cell counts. In conclusion, HTLV-1/2 LAMP had similar to superior performance than PCR-based assays, and therefore may represent an attractive alternative for HTLV-1/2 diagnosis due to reduced working time and costs, and the simple infrastructure needed.


Assuntos
Infecções por HTLV-I/virologia , Infecções por HTLV-II/virologia , Vírus Linfotrópico T Tipo 1 Humano/genética , Vírus Linfotrópico T Tipo 2 Humano/genética , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Sangue/virologia , Técnicas de Laboratório Clínico , Infecções por HTLV-I/sangue , Infecções por HTLV-I/diagnóstico , Infecções por HTLV-II/sangue , Infecções por HTLV-II/diagnóstico , Vírus Linfotrópico T Tipo 1 Humano/classificação , Vírus Linfotrópico T Tipo 1 Humano/isolamento & purificação , Vírus Linfotrópico T Tipo 2 Humano/classificação , Vírus Linfotrópico T Tipo 2 Humano/isolamento & purificação , Humanos , Sensibilidade e Especificidade
20.
Viruses ; 12(9)2020 09 04.
Artigo em Inglês | MEDLINE | ID: mdl-32899808

RESUMO

Recently, several so-called "atypical" Bluetongue virus (BTV) serotypes were discovered, including BTV-25 (Toggenburg virus), in Switzerland. Most "atypical" BTV were identified in small ruminants without clinical signs. In 2018, two goats from a holding in Germany tested positive for BTV-25 genome by RT-qPCR prior to export. After experimental inoculation of the two goats with the BTV-25 positive field blood samples for generation of reference materials, viremia could be observed in one animal. For the first time, the BTV-25-related virus was isolated in cell culture from EDTA-blood and the full genome of isolate "BTV-25-GER2018" could be generated. BTV-25-GER2018 was only incompletely neutralized by ELISA-positive sera. We could monitor the BTV-25 occurrence in the respective affected goat flock of approximately 120 goats over several years. EDTA blood samples were screened with RT-qPCR using a newly developed BTV-25 specific assay. For serological surveillance, serum samples were screened using a commercial cELISA. BTV-25-GER2018 was detected over 4.5 years in the goat flock with intermittent PCR-positivity in some animals, and with or without concomitantly detected antibodies since 2015. We could demonstrate the viral persistence of BTV-25-GER2018 in goats for up to 4.5 years, and the first BTV-25 isolate is now available for further characterization.


Assuntos
Vírus Bluetongue/isolamento & purificação , Bluetongue/virologia , Doenças das Cabras/virologia , Animais , Anticorpos Antivirais/sangue , Sangue/virologia , Bluetongue/sangue , Vírus Bluetongue/classificação , Vírus Bluetongue/genética , Vírus Bluetongue/crescimento & desenvolvimento , Genoma Viral , Doenças das Cabras/sangue , Cabras
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