RESUMO
Influenza virus is a persistent threat to human health; indeed, the deadliest modern pandemic was in 1918 when an H1N1 virus killed an estimated 50 million people globally. The intent of this work is to better understand influenza from an RNA-centric perspective to provide local, structural motifs with likely significance to the influenza infectious cycle for therapeutic targeting. To accomplish this, we analyzed over four hundred thousand RNA sequences spanning three major clades: influenza A, B and C. We scanned influenza segments for local secondary structure, identified/modeled motifs of likely functionality, and coupled the results to an analysis of evolutionary conservation. We discovered 185 significant regions of predicted ordered stability, yet evidence of sequence covariation was limited to 7 motifs, where 3-found in influenza C-had higher than expected amounts of sequence covariation.
Assuntos
Betainfluenzavirus/genética , Gammainfluenzavirus/genética , Vírus da Influenza A/genética , Estabilidade de RNA , RNA Viral/ultraestrutura , Antivirais/farmacologia , Vírus da Influenza A/efeitos dos fármacos , Betainfluenzavirus/efeitos dos fármacos , Gammainfluenzavirus/efeitos dos fármacos , Modelos Moleculares , Conformação de Ácido Nucleico , Motivos de Nucleotídeos , RNA Viral/efeitos dos fármacos , RNA Viral/genética , Análise de Sequência de RNA , Relação Estrutura-AtividadeRESUMO
Neuraminidase of influenza A and B viruses plays a critical role in the virus life cycle and is an important target of the host immune system. Here, we highlight the current understanding of influenza neuraminidase structure, function, antigenicity, immunogenicity, and immune protective potential. Neuraminidase inhibiting antibodies have been recognized as correlates of protection against disease caused by natural or experimental influenza A virus infection in humans. In the past years, we have witnessed an increasing interest in the use of influenza neuraminidase to improve the protective potential of currently used influenza vaccines. A number of well-characterized influenza neuraminidase-specific monoclonal antibodies have been described recently, most of which can protect in experimental challenge models by inhibiting the neuraminidase activity or by Fc receptor-dependent mechanisms. The relative instability of the neuraminidase poses a challenge for protein-based antigen design. We critically review the different solutions that have been proposed to solve this problem, ranging from the inclusion of stabilizing heterologous tetramerizing zippers to the introduction of inter-protomer stabilizing mutations. Computationally engineered neuraminidase antigens have been generated that offer broad, within subtype protection in animal challenge models. We also provide an overview of modern vaccine technology platforms that are compatible with the induction of robust neuraminidase-specific immune responses. In the near future, we will likely see the implementation of influenza vaccines that confront the influenza virus with a double punch: targeting both the hemagglutinin and the neuraminidase.
Assuntos
Vacinas contra Influenza/imunologia , Influenza Humana/prevenção & controle , Neuraminidase/imunologia , Proteínas Virais/imunologia , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Deriva e Deslocamento Antigênicos , Antígenos Virais/imunologia , Antígenos Virais/ultraestrutura , Domínio Catalítico/genética , Domínio Catalítico/imunologia , Proteção Cruzada , Evolução Molecular , Humanos , Imunogenicidade da Vacina , Vacinas contra Influenza/administração & dosagem , Vacinas contra Influenza/genética , Influenza Humana/imunologia , Influenza Humana/virologia , Alphainfluenzavirus/enzimologia , Alphainfluenzavirus/genética , Alphainfluenzavirus/imunologia , Betainfluenzavirus/enzimologia , Betainfluenzavirus/genética , Betainfluenzavirus/imunologia , Mutação , Nanopartículas , Neuraminidase/administração & dosagem , Neuraminidase/genética , Neuraminidase/ultraestrutura , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Vacinas Sintéticas/ultraestrutura , Proteínas Virais/administração & dosagem , Proteínas Virais/genética , Proteínas Virais/ultraestruturaRESUMO
Since the Coronavirus 19 (COVID-19) pandemic, several SARS-CoV-2 variants of concern (SARS-CoV-2 VOC) have been reported. The B.1.1.7 variant has been associated with increased mortality and transmission risk. Furthermore, cluster and possible co-infection cases could occur in the next influenza season or COVID-19 pandemic wave, warranting efficient diagnosis and treatment decision making. Here, we aimed to detect SARS-CoV-2 and other common respiratory viruses using multiplex RT-PCR developed on the LabTurbo AIO 48 open system. We performed a multicenter study to evaluate the performance and analytical sensitivity of the LabTurbo AIO 48 system for SARS-CoV-2, influenza A/B, and respiratory syncytial virus (RSV) using 652 nasopharyngeal swab clinical samples from patients. The LabTurbo AIO 48 system demonstrated a sensitivity of 9.4 copies/per PCR for N2 of SARS-CoV-2; 24 copies/per PCR for M of influenza A and B; and 24 copies/per PCR for N of RSV. The assay presented consistent performance in the multicenter study. The multiplex RT-PCR applied on the LabTurbo AIO 48 open platform provided highly sensitive, robust, and accurate results and enabled high-throughput detection of B.1.1.7, influenza A/B, and RSV with short turnaround times. Therefore, this automated molecular diagnostic assay could enable streamlined testing if COVID-19 becomes a seasonal disease.
Assuntos
Teste de Ácido Nucleico para COVID-19/métodos , COVID-19/diagnóstico , Influenza Humana/diagnóstico , Reação em Cadeia da Polimerase Multiplex/métodos , Infecções por Vírus Respiratório Sincicial/diagnóstico , Adulto , Idoso , COVID-19/virologia , Feminino , Humanos , Vírus da Influenza A/genética , Vírus da Influenza A/isolamento & purificação , Influenza Humana/virologia , Betainfluenzavirus/genética , Betainfluenzavirus/isolamento & purificação , Masculino , Pessoa de Meia-Idade , Infecções por Vírus Respiratório Sincicial/virologia , Vírus Sinciciais Respiratórios/genética , Vírus Sinciciais Respiratórios/isolamento & purificação , SARS-CoV-2/genética , SARS-CoV-2/isolamento & purificação , Sensibilidade e Especificidade , Adulto JovemRESUMO
Influenza viruses continue to be a major public health threat due to the possible emergence of more virulent influenza virus strains resulting from dynamic changes in virus adaptability, consequent of functional mutations and antigenic drift in surface proteins, especially hemagglutinin (HA) and neuraminidase (NA). In this study, we describe the genetic and evolutionary characteristics of H1N1, H3N2, and influenza B strains detected in severe cases of seasonal influenza in Thailand from 2018 to 2019. We genetically characterized seven A/H1N1 isolates, seven A/H3N2 isolates, and six influenza B isolates. Five of the seven A/H1N1 viruses were found to belong to clade 6B.1 and were antigenically similar to A/Switzerland/3330/2017 (H1N1), whereas two isolates belonged to clade 6B.1A1 and clustered with A/Brisbane/02/2018 (H1N1). Interestingly, we observed additional mutations at antigenic sites (S91R, S181T, T202I) as well as a unique mutation at a receptor binding site (S200P). Three-dimensional (3D) protein structure analysis of hemagglutinin protein reveals that this unique mutation may lead to the altered binding of the HA protein to a sialic acid receptor. A/H3N2 isolates were found to belong to clade 3C.2a2 and 3C.2a1b, clustering with A/Switzerland/8060/2017 (H3N2) and A/South Australia/34/2019 (H3N2), respectively. Amino acid sequence analysis revealed 10 mutations at antigenic sites including T144A/I, T151K, Q213R, S214P, T176K, D69N, Q277R, N137K, N187K, and E78K/G. All influenza B isolates in this study belong to the Victoria lineage. Five out of six isolates belong to clade 1A3-DEL, which relate closely to B/Washington/02/2009, with one isolate lacking the three amino acid deletion on the HA segment at position K162, N163, and D164. In comparison to the B/Colorado/06/2017, which is the representative of influenza B Victoria lineage vaccine strain, these substitutions include G129D, G133R, K136E, and V180R for HA protein. Importantly, the susceptibility to oseltamivir of influenza B isolates, but not A/H1N1 and A/H3N2 isolates, were reduced as assessed by the phenotypic assay. This study demonstrates the importance of monitoring genetic variation in influenza viruses regarding how acquired mutations could be associated with an improved adaptability for efficient transmission.
Assuntos
Betainfluenzavirus , Hospitalização , Vírus da Influenza A , Influenza Humana/epidemiologia , Influenza Humana/virologia , Adolescente , Adulto , Idoso , Antígenos Virais/química , Antígenos Virais/imunologia , Antivirais/farmacologia , Antivirais/uso terapêutico , Comorbidade , Feminino , História do Século XXI , Humanos , Vírus da Influenza A/classificação , Vírus da Influenza A/efeitos dos fármacos , Vírus da Influenza A/genética , Influenza Humana/tratamento farmacológico , Influenza Humana/história , Betainfluenzavirus/classificação , Betainfluenzavirus/efeitos dos fármacos , Betainfluenzavirus/genética , Masculino , Pessoa de Meia-Idade , Modelos Moleculares , Neuraminidase/química , Neuraminidase/imunologia , Neuraminidase/metabolismo , Filogenia , Estações do Ano , Tailândia/epidemiologia , Proteínas Virais/química , Proteínas Virais/imunologia , Proteínas Virais/metabolismo , Adulto JovemRESUMO
BACKGROUND: Diagnosis of influenza in patients admitted to hospital is delayed due to long turnaround times with laboratory testing, leading to inappropriate and late antiviral treatment and isolation facility use. Molecular point-of-care tests (mPOCTs) are highly accurate, easy to use, and generate results in less than 1 h, but high-quality evidence for their effect on management and clinical outcomes is needed. The aim of this study was to assess the clinical impact of an mPOCT on influenza detection, antiviral use, infection control measures, and clinical outcomes in adults admitted to hospital with acute respiratory illness. METHODS: In this multicentre, pragmatic, open-label, randomised controlled trial (FluPOC), we recruited adults admitted to hospital with acute respiratory illness during influenza seasons from two hospitals in Hampshire, UK. Eligible patients were aged 18 years and older, with acute respiratory illness of 10 days or fewer duration before admission to hospital, who were recruited within 16 h of admission to hospital. Participants were randomly assigned (1:1), using random permuted blocks of varying sizes (4, 6 and 8), to receive mPOCT for influenza or routine clinical care (control group). The primary outcome was the proportion of patients infected with influenza who were treated appropriately with antivirals (neuraminidase inhibitors) within 5 days of admission. Safety was assessed in all patients. Secondary outcomes included time to antivirals, isolation facility use, and clinical outcomes. This study is registered with the ISRCTN registry, ISRCTN17197293, and is now complete. FINDINGS: Between Dec 12, 2017, and May 3, 2019, over two influenza seasons, 613 patients were enrolled, of whom 307 were assigned to the mPOCT group and 306 to the control group, and all were analysed. Median age was 62 years (IQR 45-75) and 332 (54%) of 612 participants with data were female. 100 (33%) of 307 patients in the mPOCT group and 102 (33%) of 306 in the control group had influenza. 100 (100%) of 100 patients with influenza were diagnosed in the mPOCT group and 60 (59%) of 102 were diagnosed though routine clinical care in the control group (relative risk 1·7, 95% CI 1·7-1·7; p<0·0001). 99 (99%) of 100 patients with influenza in the mPOCT group were given antiviral treatment within 5 days of admission versus 63 (62%) 102 in the control group (relative risk 1·6, 95% CI 1·4-1·9; p<0·0001). Median time to antivirals was 1·0 h (IQR 0·0 to 2·0) in the mPOCT group versus 6·0 h (0·0 to 12·0) in the control group (difference of 5·0 h [95% CI 0·0-6·0; p=0·0039]). 70 (70%) of 100 patients with influenza in the mPOCT group were isolated to single-room accommodation versus 39 (38%) of 102 in the control group (relative risk 1·8 [95% CI 1·4-2·4; p<0·0001]). 19 adverse events occurred among patients with influenza in the mPOCT group compared with 34 events in the control group. No patients with influenza died in the mPOCT group and two (2%) died in the control group (p=0·16). INTERPRETATION: Routine mPOCT for influenza was associated with improved influenza detection and improvements in appropriate and timely antiviral and isolation facility use. Routine mPOCT should replace laboratory-based diagnostics for acute admissions to hospital during the influenza season. FUNDING: National Institute for Health Research.
Assuntos
Antivirais/uso terapêutico , Controle de Infecções/organização & administração , Influenza Humana/diagnóstico , Técnicas de Diagnóstico Molecular/instrumentação , Testes Imediatos/organização & administração , Idoso , Feminino , Humanos , Controle de Infecções/estatística & dados numéricos , Influenza Humana/tratamento farmacológico , Influenza Humana/epidemiologia , Influenza Humana/virologia , Alphainfluenzavirus/genética , Alphainfluenzavirus/isolamento & purificação , Betainfluenzavirus/genética , Betainfluenzavirus/isolamento & purificação , Tempo de Internação/estatística & dados numéricos , Masculino , Pessoa de Meia-Idade , Técnicas de Diagnóstico Molecular/métodos , Admissão do Paciente , Reação em Cadeia da Polimerase , RNA Viral/isolamento & purificação , Fatores de Tempo , Tempo para o Tratamento/estatística & dados numéricos , Resultado do TratamentoRESUMO
BACKGROUND: Influenza B is often perceived as a less severe strain of influenza. The epidemiology and clinical outcomes of influenza B have been less thoroughly investigated in hospitalised patients. The aims of this study were to describe clinical differences and outcomes between influenza A and B patients admitted over a period of 4 years. METHODS: We retrospectively collected data of all laboratory confirmed influenza patients ≥18 years at two tertiary hospitals in South Australia. Patients were confirmed as influenza positive if they had a positive polymerase-chain-reaction (PCR) test of a respiratory specimen. Complications during hospitalisation along with inpatient mortality were compared between influenza A and B. In addition, 30 day mortality and readmissions were compared. Logistic regression model compared outcomes after adjustment for age, Charlson index, sex and creatinine levels. RESULTS: Between January 2016-March 2020, 1846 patients, mean age 66.5 years, were hospitalised for influenza. Of whom, 1630 (88.3%) had influenza A and 216 (11.7%) influenza B. Influenza B patients were significantly younger than influenza A. Influenza A patients were more likely be smokers with a history of chronic obstructive pulmonary disease (COPD) and ischaemic heart disease (IHD) than influenza B. Complications, including pneumonia and acute coronary syndrome (ACS) were similar between two groups, however, septic shock was more common in patients with influenza B. Adjusted analyses showed similar median length of hospital stay (LOS), in hospital mortality, 30-day mortality and readmissions between the two groups. CONCLUSIONS: Influenza B is less prevalent and occurs mostly in younger hospitalised patients than influenza A. Both strains contribute equally to hospitalisation burden and complications. TRIAL REGISTRATION: Australia and New Zealand Clinical Trial Registry (ANZCR) no ACTRN12618000451202 date of registration 28/03/2018.
Assuntos
Betainfluenzavirus/genética , Vírus da Influenza A/genética , Influenza Humana/epidemiologia , Influenza Humana/mortalidade , Síndrome Coronariana Aguda/etiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Austrália/epidemiologia , Feminino , Mortalidade Hospitalar , Humanos , Influenza Humana/complicações , Influenza Humana/virologia , Tempo de Internação , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Readmissão do Paciente , Pneumonia/etiologia , Reação em Cadeia da Polimerase , Prevalência , Estudos RetrospectivosRESUMO
OBJECTIVE: The aim of this study is to find the distributions of pathogens in 164 suspected COVID-19 patients from the outpatient clinic of Shenjing Hospital of China Medical University from 24th January, 2020, to 29th February of 2020. PATIENTS AND METHODS: 164 COVID-19 suspected patients were from the Shengjing Hospital of China Medical University. Oropharyngeal swab specimens were acquired by respiratory doctors under standardized conditions. Specific nucleic acids of SARS-CoV-2, influenza A and B, respiratory syncytial virus A and B, adenovirus, parainfluenza virus, along with pneumonic mycoplasma were detected by real-time fluorescence PCR. Symptomatic, epidemiologic, laboratory and radiological data of the patients were obtained from the electronic medical record system of our hospital. RESULTS: Among the 164 patients, 3 were positive for SARS-CoV-2, 15 were positive for other respiratory viruses and 16 were positive for pneumonic mycoplasma. Of the positive patients above, 1 patient was co-infected with SARS-CoV-2 and adenovirus, and 1 was co-infected with influenza B and pneumonic mycoplasma. The 3 SARS-CoV-2 infected patients were clinically diagnosed as COVID-19 because they meet the diagnostic criteria listed in "Chinese Clinical Guidance for COVID-19 Pneumonia diagnosis and treatment", including epidemic history, symptom and pathogenic detection, as well as abnormalities of the laboratory and radiological data. However, the clinical characteristics of COVID-19 patients were non-specific compared to those of the patients infected with other respiratory viruses. CONCLUSIONS: The endemic common respiratory pathogens are more prevalent than SARS-CoV-2 in the SARS-CoV-2 non-epidemic areas of this research. Detection of the pathogen is the unique means for definite COVID-19 diagnosis.
Assuntos
Infecções por Coronavirus/patologia , Pneumonia Viral/patologia , Adenoviridae/genética , Adenoviridae/isolamento & purificação , Betacoronavirus/genética , Betacoronavirus/isolamento & purificação , COVID-19 , China/epidemiologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Humanos , Vírus da Influenza A/genética , Vírus da Influenza A/isolamento & purificação , Betainfluenzavirus/genética , Betainfluenzavirus/isolamento & purificação , Pandemias , Pneumonia Viral/epidemiologia , Pneumonia Viral/virologia , RNA Viral/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , SARS-CoV-2 , Tórax/diagnóstico por imagem , Tomografia Computadorizada por Raios XRESUMO
INTRODUCTION: Every winter, emergency departments (EDs) face overcrowding with patients presenting influenza-like symptoms, and organisational issues such as single room assignment and droplet precautions to avoid hospital-acquired influenza. Our main objective was to assess the impact of PCR results and patient's severity on single room assignment. METHODS: All patients admitted to Bichat-Claude Bernard Hospital through the ED and tested for influenza by PCR (GenXpert, Cepheid) or (FilmArray, BioMérieux) on a nasopharyngeal swab were retrospectively included during three influenza seasons (2015-2018. RESULTS: Of 1,330 included patients, 278 (20.9%) had a positive PCR for influenza. The median time to obtain a PCR result was 19 hours, and 238 (18.3%) patients were assigned a single room. Among patients with positive and negative influenza PCR, 22.3% and 16.7% were assigned a single room (p = 0.03). The multivariable analysis was performed on the two first epidemic periods, excluding the third epidemic because of the concomitant use of influenza immune-chromatic test. Only level 1 of the Emergency severity index (ESI) (aOR, 1.9; 95% CI, 1.3-2.8; p < 0.01) was associated with single-room assignment. PCR result was not statistically associated with the decision of single room assignment (aOR, 1.4; 95%CI, 1.0-1.4; p = 0.07). CONCLUSION: A PCR positive for influenza was not significantly associated with single-room assignment. Less than one quarter of influenza patients were adequately assigned a single room, likely due to the long turnaround time of PCR result and other conflicting indications for single room-assignment. Accelerating biological diagnosis could improve single-room assignment.
Assuntos
Betainfluenzavirus/isolamento & purificação , Serviço Hospitalar de Emergência/organização & administração , Vírus da Influenza A/isolamento & purificação , Influenza Humana/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Testes Diagnósticos de Rotina , Feminino , Humanos , Vírus da Influenza A/genética , Influenza Humana/epidemiologia , Betainfluenzavirus/genética , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Estudos RetrospectivosRESUMO
BACKGROUND: Since 2011, the Global Influenza Hospital Surveillance Network (GIHSN) has used active surveillance to prospectively collect epidemiological and virological data on patients hospitalized with influenza virus infection. Here, we describe influenza virus strain circulation in the GIHSN participant countries during 2017-2018 season and examine factors associated with complicated hospitalization among patients admitted with laboratory-confirmed influenza illness. METHODS: The study enrolled patients who were hospitalized in a GIHSN hospital in the previous 48 h with acute respiratory symptoms and who had symptoms consistent with influenza within the 7 days before admission. Enrolled patients were tested by reverse transcription-polymerase chain reaction to confirm influenza virus infection. "Complicated hospitalization" was defined as a need for mechanical ventilation, admission to an intensive care unit, or in-hospital death. In each of four age strata (< 15, 15-< 50, 50-< 65, and ≥ 65 years), factors associated with complicated hospitalization in influenza-positive patients were identified by mixed effects logistic regression and those associated with length of hospital stay using a linear mixed-effects regression model. RESULTS: The study included 12,803 hospitalized patients at 14 coordinating sites in 13 countries, of which 4306 (34%) tested positive for influenza. Influenza viruses B/Yamagata, A/H3N2, and A/H1N1pdm09 strains dominated and cocirculated, although the dominant strains varied between sites. Complicated hospitalization occurred in 10.6% of influenza-positive patients. Factors associated with complicated hospitalization in influenza-positive patients included chronic obstructive pulmonary disease (15-< 50 years and ≥ 65 years), diabetes (15-< 50 years), male sex (50-< 65 years), hospitalization during the last 12 months (50-< 65 years), and current smoking (≥65 years). Chronic obstructive pulmonary disease (50-< 65 years), other chronic conditions (15-< 50 years), influenza A (50-< 65 years), and hospitalization during the last 12 months (< 15 years) were associated with a longer hospital stay. The proportion of patients with complicated influenza did not differ between influenza A and B. CONCLUSIONS: Complicated hospitalizations occurred in over 10% of patients hospitalized with influenza virus infection. Factors commonly associated with complicated or longer hospitalization differed by age group but commonly included chronic obstructive pulmonary disease, diabetes, and hospitalization during the last 12 months.
Assuntos
Betainfluenzavirus/genética , Hospitalização , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H3N2/genética , Influenza Humana/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Feminino , Mortalidade Hospitalar , Humanos , Lactente , Influenza Humana/mortalidade , Influenza Humana/virologia , Unidades de Terapia Intensiva , Tempo de Internação , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Respiração Artificial , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Risco , Adulto JovemRESUMO
In the setting of the coronavirus disease 2019 (COVID-19) pandemic, only few data regarding lung pathology induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is available, especially without medical intervention interfering with the natural evolution of the disease. We present here the first case of forensic autopsy of a COVID-19 fatality occurring in a young woman, in the community. Diagnosis was made at necropsy and lung histology showed diffuse alveolar damage, edema, and interstitial pneumonia with a geographically heterogeneous pattern, mostly affecting the central part of the lungs. This death related to COVID-19 pathology highlights the heterogeneity and severity of central lung lesions after natural evolution of the disease.
Assuntos
Betacoronavirus , Infecções por Coronavirus/patologia , Pulmão/patologia , Pneumonia Viral/patologia , Adenoviridae/genética , Adenoviridae/isolamento & purificação , Adulto , Autopsia , Betacoronavirus/genética , Betacoronavirus/isolamento & purificação , Bocavirus/genética , Bocavirus/isolamento & purificação , Proteína C-Reativa/análise , COVID-19 , Coronavirus/genética , Coronavirus/isolamento & purificação , Feminino , Humanos , Alphainfluenzavirus/genética , Alphainfluenzavirus/isolamento & purificação , Betainfluenzavirus/genética , Betainfluenzavirus/isolamento & purificação , Macrófagos/patologia , Megacariócitos/patologia , Metapneumovirus/genética , Metapneumovirus/isolamento & purificação , Neutrófilos/patologia , Obesidade Mórbida , Pandemias , Pró-Calcitonina/sangue , Reação em Cadeia da Polimerase em Tempo Real , Vírus Sincicial Respiratório Humano/genética , Vírus Sincicial Respiratório Humano/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rhinovirus/genética , Rhinovirus/isolamento & purificação , SARS-CoV-2 , Suíça , Linfócitos T/patologiaRESUMO
BACKGROUND: This study compared the genomes of influenza viruses that caused mild infections among outpatients and severe infections among hospitalized patients in Singapore, and characterized their molecular evolution and receptor-binding specificity. METHODS: The complete genomes of influenza A/H1N1, A/H3N2 and B viruses that caused mild infections among outpatients and severe infections among inpatients in Singapore during 2012-2015 were sequenced and characterized. Using various bioinformatics approaches, we elucidated their evolutionary, mutational and structural patterns against the background of global and vaccine strains. RESULTS: The phylogenetic trees of the 8 gene segments revealed that the outpatient and inpatient strains overlapped with representative global and vaccine strains. We observed a cluster of inpatients with A/H3N2 strains that were closely related to vaccine strain A/Texas/50/2012(H3N2). Several protein sites could accurately discriminate between outpatient versus inpatient strains, with site 221 in neuraminidase (NA) achieving the highest accuracy for A/H3N2. Interestingly, amino acid residues of inpatient but not outpatient isolates at those sites generally matched the corresponding residues in vaccine strains, except at site 145 of hemagglutinin (HA). This would be especially relevant for future surveillance of A/H3N2 strains in relation to their antigenicity and virulence. Furthermore, we observed a trend in which the HA proteins of influenza A/H3N2 and A/H1N1 exhibited enhanced ability to bind both avian and human host cell receptors. In contrast, the binding ability to each receptor was relatively stable for the HA of influenza B. CONCLUSIONS: Overall, our findings extend our understanding of the molecular and structural evolution of influenza virus strains in Singapore within the global context of these dynamic viruses.
Assuntos
Betainfluenzavirus/genética , Evolução Molecular , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H3N2/genética , Adolescente , Adulto , Idoso , Glicoproteínas de Hemaglutininação de Vírus da Influenza/química , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Hospitalização , Humanos , Influenza Humana/virologia , Pessoa de Meia-Idade , Mutação , Neuraminidase/genética , Pacientes Ambulatoriais , Filogenia , Receptores Virais/química , Singapura , Proteínas Virais/genética , Adulto JovemRESUMO
We studied the constellation of genes encoding polymerase complex proteins of master donor viruses for Russian live attenuated influenza vaccine type B. Reassortants of the reserve attenuation donor B/Leningrad/14/17/55 with B/USSR/60/69 master donor virus currently used for manufacturing seasonal influenza vaccine were prepared and examined. Most reassortants obtained by the classical reassortment method inherited all genes from the B/Leningrad/14/17/55 virus except the gene encoding PB1 subunit of the polymerase complex. One reassortant was selected for further evaluation of the role of PB1 gene. Greater attenuation of the strain for experimental animals (mice) in comparison with the original strains was demonstrated. This indicates high degree of constellation of genes of cold-adapted master donor viruses and the important compensating role of amino acid substitutions in the PB1 protein of B/Leningrad/14/17/55 donor virus in preventing viral hyperattenuation.
Assuntos
Betainfluenzavirus/genética , Influenza Humana/prevenção & controle , Proteínas Virais/genética , Substituição de Aminoácidos/genética , Animais , Embrião de Galinha , Temperatura Baixa , Humanos , Vacinas contra Influenza/imunologia , Betainfluenzavirus/imunologia , Camundongos , Vírus Reordenados/genética , Vírus Reordenados/imunologia , Vacinas Atenuadas/imunologiaRESUMO
Influenza virus RNA-dependent RNA polymerase uses unique mechanisms to transcribe its single-stranded genomic viral RNA (vRNA) into messenger RNA. The polymerase is initially bound to a promoter comprising the partially base-paired 3' and 5' extremities of the RNA. A short, capped primer, 'cap-snatched' from a nascent host polymerase II transcript, is directed towards the polymerase active site to initiate RNA synthesis. Here we present structural snapshots, as determined by X-ray crystallography and cryo-electron microscopy, of actively initiating influenza polymerase as it transitions towards processive elongation. Unexpected conformational changes unblock the active site cavity to allow establishment of a nine-base-pair template-product RNA duplex before the strands separate into distinct exit channels. Concomitantly, as the template translocates, the promoter base pairs are broken and the template entry region is remodeled. These structures reveal details of the influenza polymerase active site that will help optimize nucleoside analogs or other compounds that directly inhibit viral RNA synthesis.
Assuntos
Betainfluenzavirus/enzimologia , RNA Polimerase Dependente de RNA/química , Proteínas Virais/química , Domínio Catalítico , Cristalografia por Raios X , Humanos , Influenza Humana/virologia , Betainfluenzavirus/química , Betainfluenzavirus/genética , Betainfluenzavirus/metabolismo , Modelos Moleculares , Conformação Proteica , RNA Viral/química , RNA Viral/genética , RNA Viral/metabolismo , RNA Polimerase Dependente de RNA/metabolismo , Elongação da Transcrição Genética , Iniciação da Transcrição Genética , Proteínas Virais/metabolismoRESUMO
BACKGROUND: Although Indonesia has high fatality rate of human A/H5N1 cases, epidemiological and clinical data on influenza virus circulation among humans has been limited. Within Indonesia, Bali province is of interest due to high population densities of humans, pigs and poultry. This study aims to characterize and compare the epidemiological and clinical patterns of influenza viruses in humans through surveillance among patients with influenza-like illness (ILI) in Bali, Indonesia. METHODS: ILI patients were recruited at 21 sentinel health facilities across all nine regencies in Bali, from July 2010 to June 2014. PCR-based assays were used for detection and subtyping of influenza viruses. Demographic, behavioural and clinical data were tested for associations with influenza using chi-squared tests and logistic regression. RESULTS: Of 2077 ILI patients, 291 (14.0%) tested positive for influenza A, 152 (7.3%) for influenza B, and 16 (0.77%) for both influenza A and B. Of the influenza A isolates, the majority 61.2% were A/H3N2, followed by A/H1N1-pdm09 (80; 26.1%). Two A/H5N1 were identified. Influenza positive rates were significantly higher during wet season months (28.3%), compared with the dry season (13.8%; χ2 = 61.1; df = 1; p < 0.0001). Clinical predictors for infection varied by virus type, with measured fever (≥38 °C) more strongly associated with influenza B (AOR: 1.62; 95% CI: 1.10, 2.39). CONCLUSION: Influenza circulates year-round among humans in Bali with higher activity during the wet season. High contact rates with poultry and pigs, along with influenza virus detection that could not be subtyped through conventional assays, highlight the need for molecular studies to characterize epidemiological and evolutionary dynamics of influenza in this setting.
Assuntos
Betainfluenzavirus/genética , Vírus da Influenza A/genética , Influenza Humana/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Feminino , Humanos , Indonésia/epidemiologia , Lactente , Recém-Nascido , Vírus da Influenza A/isolamento & purificação , Influenza Humana/epidemiologia , Influenza Humana/virologia , Betainfluenzavirus/isolamento & purificação , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Estações do Ano , Adulto JovemRESUMO
The World Health Organization selects influenza vaccine compositions biannually to cater to peaks in temperate regions. In tropical and subtropical regions, where influenza seasonality varies and epidemics can occur year-round, the choice of vaccine remains uncertain. Our 17-year molecular epidemiologic survey showed that most influenza A(H3N2) (9/11) and B (6/7) vaccine strains had circulated in East Asia >1 year before inclusion into vaccines. Northern Hemisphere vaccine strains and circulating strains in East Asia were closely matched in 7 (20.6%) of 34 seasons for H3N2 and 5 (14.7%) of 34 seasons for B. Southern Hemisphere vaccines also had a low probability of matching (H3N2, 14.7%; B, 11.1%). Strain drift among seasons was common (H3N2, 41.2%; B, 35.3%), and biannual vaccination strategy (Northern Hemisphere vaccines in November followed by Southern Hemisphere vaccines in May) did not improve matching. East Asia is an important contributor to influenza surveillance but often has mismatch between vaccine and contemporarily circulating strains.
Assuntos
Alphainfluenzavirus/genética , Betainfluenzavirus/genética , Variação Genética , Vacinas contra Influenza/genética , Influenza Humana/epidemiologia , Influenza Humana/virologia , Estações do Ano , Glicoproteínas de Hemaglutininação de Vírus da Influenza/química , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , História do Século XX , História do Século XXI , Hong Kong/epidemiologia , Humanos , Vacinas contra Influenza/imunologia , Influenza Humana/história , Influenza Humana/prevenção & controle , Alphainfluenzavirus/classificação , Alphainfluenzavirus/imunologia , Betainfluenzavirus/classificação , Betainfluenzavirus/imunologia , Epidemiologia Molecular , Filogenia , RNA Viral , Estudos RetrospectivosRESUMO
We present a case involving an 85-year-old man with acute confusion and new onset seizure following a 1-week history of respiratory prodrome. This case report describes a case of influenza B-related meningoencephalitis supported by evidence of an influenza B infection and temporal relation of the neurological event and respiratory illness in the absence of other identifiable cause. Diagnosis is guided by cerebrospinal fluid profile and nasopharyngeal PCR. Treatment is largely supportive and the effect of vaccination on prevention of this neurological complication remains unclear.
Assuntos
Antivirais/uso terapêutico , Betainfluenzavirus/isolamento & purificação , Influenza Humana/diagnóstico , Influenza Humana/tratamento farmacológico , Meningoencefalite/virologia , Idoso de 80 Anos ou mais , Confusão/etiologia , Humanos , Influenza Humana/líquido cefalorraquidiano , Betainfluenzavirus/genética , Levetiracetam , Masculino , Meningoencefalite/líquido cefalorraquidiano , Meningoencefalite/tratamento farmacológico , Nasofaringe/virologia , Piracetam/análogos & derivados , Piracetam/uso terapêutico , Convulsões/tratamento farmacológico , Convulsões/etiologia , Resultado do Tratamento , Ácido Valproico/uso terapêuticoRESUMO
INTRODUCTION: The Alere i Influenza A & B assay incorporates the Nicking Enzyme Amplification Reaction technique on the Alere i instrument to detect and differentiate influenza virus (Flu) A and B nucleic acids in specific specimens. Areas covered: The Alere i Influenza A & B assay was cleared by the US Food and Drug Administration for use with nasal swabs (NS) and nasopharyngeal swabs, either directly or in viral transport medium. Notably, direct use on NS was the first ever CLIA-waived nucleic acid-based test. Previously published evaluations have reported sensitivities and specificities of 55.2-100% and 62.5-100% for Flu A and 45.2-100% and 53.6-100% for Flu B, respectively. Expert commentary: The Alere i Influenza A & B assay provides a rapid and simple platform for detection and differentiation of Flu A and B. Efforts are expected to further improve sensitivity and user-friendliness for effective and widespread use in the true point-of-care setting.
Assuntos
Betainfluenzavirus/genética , Vírus da Influenza A/genética , Influenza Humana/diagnóstico , Influenza Humana/virologia , Técnicas de Diagnóstico Molecular , Testes Imediatos , Humanos , Técnicas de Amplificação de Ácido Nucleico , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
The clinical effectiveness of four neuraminidase inhibitors (NAIs) (oseltamivir, zanamivir, laninamivir, and peramivir) for children aged 0 months to 18 years with influenza A and B were investigated in the 2014-2015 to 2016-2017 influenza seasons in Japan. A total of 1207 patients (747 with influenza A and 460 with influenza B) were enrolled. The Cox proportional-hazards model using all of the patients showed that the duration of fever after administration of the first dose of the NAI was shorter in older patients (hazard ratio = 1.06 per 1 year of age, p < 0.001) and that the duration of fever after administration of the first dose of the NAI was shorter in patients with influenza A infection than in patients with influenza B infection (hazard ratio = 2.21, p < 0.001). A logistic regression model showed that the number of biphasic fever episodes was 2.99-times greater for influenza B-infected patients than for influenza A-infected patients (p < 0.001). The number of biphasic fever episodes in influenza A- or B-infected patients aged 0-4 years was 2.89-times greater than that in patients aged 10-18 years (p = 0.010), and the number of episodes in influenza A- or B-infected patients aged 5-9 years was 2.13-times greater than that in patients aged 10-18 years (p = 0.012).
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Ciclopentanos/administração & dosagem , Inibidores Enzimáticos/administração & dosagem , Guanidinas/administração & dosagem , Influenza Humana/tratamento farmacológico , Neuraminidase/antagonistas & inibidores , Oseltamivir/administração & dosagem , Zanamivir/análogos & derivados , Zanamivir/administração & dosagem , Ácidos Carbocíclicos , Adolescente , Criança , Pré-Escolar , Ciclopentanos/uso terapêutico , Inibidores Enzimáticos/uso terapêutico , Feminino , Guanidinas/uso terapêutico , Humanos , Lactente , Recém-Nascido , Vírus da Influenza A/efeitos dos fármacos , Vírus da Influenza A/genética , Betainfluenzavirus/efeitos dos fármacos , Betainfluenzavirus/genética , Japão , Masculino , Oseltamivir/uso terapêutico , Piranos , Estações do Ano , Ácidos Siálicos , Resultado do Tratamento , Zanamivir/uso terapêuticoRESUMO
An increase in hospital admissions for influenza occurred during the summer of 2015 at an acute care facility in Vancouver, British Columbia, Canada. Investigation identified 25 patients with recent history of cruise ship travel to Alaska. All characterized influenza A viruses were A(H3N2). We describe patient treatment regimens and outcomes.