RESUMO
Murine hybridomas producing neutralizing mAbs specific to the pandemic influenza virus A/California/07/2009 haemagglutinin (HA) were isolated. These antibodies recognized at least two different but overlapping new epitopes that were conserved in the HA of most Spanish pandemic isolates. However, one of these isolates (A/Extremadura/RR6530/2010) lacked reactivity with the mAbs and carried two unique mutations in the HA head (S88Y and K136N) that were required simultaneously to eliminate reactivity with the murine antibodies. This unusual requirement directly illustrates the phenomenon of enhanced antigenic change proposed previously for the accumulation of simultaneous amino acid substitutions at antigenic sites of the influenza A virus HA during virus evolution (Shih et al., Proc Natl Acad Sci USA, 104â, 6283-6288, 2007). The changes found in the A/Extremadura/RR6530/2010 HA were not found in escape mutants selected in vitro with one of the mAbs, which contained instead nearby single amino acid changes in the HA head. Thus, either single or double point mutations may similarly alter epitopes of the new antigenic site identified in this work in the 2009 H1N1 pandemic virus HA. Moreover, this site is relevant for the human antibody response, as shown by competition of mAbs and human post-infection sera for virus binding. The results are discussed in the context of the HA antigenic structure and challenges posed for identification of sequence changes with possible antigenic impact during virus surveillance.
Assuntos
Antígenos Virais/genética , Antígenos Virais/imunologia , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H1N1/imunologia , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Antivirais/imunologia , Epitopos/genética , Epitopos/imunologia , Humanos , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Influenza Humana/virologia , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Proteínas Mutantes/genética , Proteínas Mutantes/imunologia , Mutação de Sentido Incorreto , RNA Viral/genética , Análise de Sequência de DNA , EspanhaRESUMO
OBJECTIVES: This study aimed to develop a genotypic method to predict HIV-1 coreceptor usage by employing the nucleotide sequence of the env gene in a tree-augmented naive Bayes (TAN) classifier, and to evaluate its accuracy in prediction compared with other available tools. METHODS: A wrapper data-mining strategy interleaved with a TAN algorithm was employed to evaluate the predictor value of every single-nucleotide position throughout the HIV-1 env gene. Based on these results, different nucleotide positions were selected to develop a TAN classifier, which was employed to predict the coreceptor tropism of all the full-length env gene sequences with information on coreceptor tropism currently available at the Los Alamos HIV Sequence Database. RESULTS: Employing 26 nucleotide positions in the TAN classifier, an accuracy of 95.6%, a specificity (identification of CCR5-tropic viruses) of 99.4% and a sensitivity (identification of CXCR4/dual-tropic viruses) of 80.5% were achieved for the in silico cross-validation. Compared with the phenotypic determination of coreceptor usage, the TAN algorithm achieved more accurate predictions than WebPSSM and Geno2pheno [coreceptor] (P<0.05). CONCLUSIONS: The use of the methodology presented in this work constitutes a robust strategy to identify genetic patterns throughout the HIV-1 env gene differently present in CCR5-tropic and CXCR4/dual-tropic viruses. Moreover, the TAN classifier can be used as a genotypic tool to predict the coreceptor usage of HIV-1 isolates reaching more accurate predictions than with other widely used genotypic tools. The use of this algorithm could improve the correct prescribing of CCR5 antagonist drugs to HIV-1-infected patients.
Assuntos
HIV-1/genética , Técnicas de Diagnóstico Molecular/métodos , Receptores de HIV/metabolismo , Tropismo Viral , Virologia/métodos , Produtos do Gene env do Vírus da Imunodeficiência Humana/genética , Biologia Computacional/métodos , Genótipo , HIV-1/fisiologia , HumanosRESUMO
We present major results concerning isolation and determination of the nucleotide sequence of hemagglutinin (HA1) of the pandemic (H1N1)pdm09 influenza viruses found in Tunisia. Amino acid analysis revealed minor amino acid changes in the antigenic or receptor-binding domains. We found mutations that were also present in 1918 pandemic virus, which includes S183P in 4 and S185T mutation in 19 of 27 viruses analyzed from 2011, while none of the 2009 viruses carried these mutations. Also two specific amino acid differences into N-glycosylation sites (N288T and N276H) were detected. The phylogenetic analysis revealed that the majority of the Tunisian isolates clustered with clade A/St. Petersburg/27/2011 viruses characterized by D97N and S185T mutations. However it also reveals a trend of 2010 strains to accumulate amino acid variation and form new phylogenetic clade with three specific amino acid substitutions: V47I, E172K and K308E.
Assuntos
Variação Genética , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Vírus da Influenza A Subtipo H1N1/genética , Influenza Humana/virologia , Substituição de Aminoácidos , Análise por Conglomerados , Humanos , Dados de Sequência Molecular , Filogenia , RNA Viral/genética , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , TunísiaRESUMO
Recently, the D222G substitution was observed in the HA of pandemic (H1N1) 2009 viruses isolated from fatal cases in several countries. We made a similar observation in one fatal case in Tunisia showing a D222G substitution in a virus isolate. The man was 47 years old and had no other subjacent pathologies or any known risk factors. He died after three days, suffering from severe respiratory symptoms of flu. The causal link of the D222G substitution in Tunisia with virulence must be verified. Further study is warranted to elucidate the intriguing relationship between the D222G substitution and severe disease. Constant molecular surveillance is important to monitor the pathogenicity of circulating strains and evaluate vaccine efficacy.
Assuntos
Hemaglutininas Virais/genética , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Influenza Humana/virologia , Mutação de Sentido Incorreto , Substituição de Aminoácidos , Evolução Fatal , Humanos , Vírus da Influenza A Subtipo H1N1/genética , Influenza Humana/mortalidade , Influenza Humana/patologia , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Proteínas Mutantes/genética , RNA Viral/genética , Análise de Sequência de DNA , TunísiaRESUMO
BACKGROUND: To estimate the prevalence of recent infection (RI) among people newly diagnosed with HIV in Spain using a representative sample collected by the AIDS Research Network cohort (CoRIS) during 2015-2016. METHODS: Stratified sampling of CoRIS data was used with proportional allocation by mode of transmission of new HIV diagnoses notified to National Surveillance System. Samples used were from patients in the CoRIS cohort with available stored plasma collected within 6 months after diagnosis. Weighted methods were used to estimate the prevalence of RI and multivariate logistic regression models were used to determine associated factors. RESULTS: Of the 669 individuals included, 55.1% were men who had sex with men (MSM), 24.6% were heterosexual, and 20.3% were non-MSM non-heterosexual. The weighted prevalence of RI was 11.8% (95% Confidence interval [CI] 9.4-14.8%) overall, 15.5% (12.2-19.4%) among MSM, 6.3% (3.9-10.0%) among heterosexual, and 8.6% (3.2-20.9%) in non-MSM non-heterosexual persons. Factors associated with prevalence of RI were: MSM (OR 2.05; 95% CI 1.02-4.14) vs. heterosexual, being Spanish (OR 2.92; 1.36-6.26) or European (OR 3.42; 1.28-9.13) vs. Latin American, having a secondary or higher education level (OR 3.08; 0.95-1.00) vs. primary, and having a CD4 count of 350-499 (OR 3.26; 1.46-7.30) or >500 (OR 6.26; 2.92-13.39) vs. <350 cells/mm3. CONCLUSIONS: In the absence of direct data from surveillance systems, the use of cohort data is a very valuable option for identifying the prevalence of RI at national level. This is the first nationwide study carried out in Spain to determine the prevalence of RI using an avidity assay.
Assuntos
Infecções por HIV , Minorias Sexuais e de Gênero , Contagem de Linfócito CD4 , Feminino , Infecções por HIV/complicações , Infecções por HIV/diagnóstico , Infecções por HIV/epidemiologia , Homossexualidade Masculina , Humanos , Masculino , Espanha/epidemiologiaRESUMO
In Western Europe, the HIV-1 epidemic among men who have sex with men (MSM) is dominated by subtype B. However, recently, other genetic forms have been reported to circulate in this population, as evidenced by their grouping in clusters predominantly comprising European individuals. Here we describe four large HIV-1 non-subtype B clusters spreading among MSM in Spain. Samples were collected in 9 regions. A pol fragment was amplified from plasma RNA or blood-extracted DNA. Phylogenetic analyses were performed via maximum likelihood, including database sequences of the same genetic forms as the identified clusters. Times and locations of the most recent common ancestors (MRCA) of clusters were estimated with a Bayesian method. Five large non-subtype B clusters associated with MSM were identified. The largest one, of F1 subtype, was reported previously. The other four were of CRF02_AG (CRF02_1; n = 115) and subtypes A1 (A1_1; n = 66), F1 (F1_3; n = 36), and C (C_7; n = 17). Most individuals belonging to them had been diagnosed of HIV-1 infection in the last 10 years. Each cluster comprised viruses from 3 to 8 Spanish regions and also comprised or was related to viruses from other countries: CRF02_1 comprised a Japanese subcluster and viruses from 8 other countries from Western Europe, Asia, and South America; A1_1 comprised viruses from Portugal, United Kingom, and United States, and was related to the A1 strain circulating in Greece, Albania and Cyprus; F1_3 was related to viruses from Romania; and C_7 comprised viruses from Portugal and was related to a virus from Mozambique. A subcluster within CRF02_1 was associated with heterosexual transmission. Near full-length genomes of each cluster were of uniform genetic form. Times of MRCAs of CRF02_1, A1_1, F1_3, and C_7 were estimated around 1986, 1989, 2013, and 1983, respectively. MRCA locations for CRF02_1 and A1_1 were uncertain (however initial expansions in Spain in Madrid and Vigo, respectively, were estimated) and were most probable in Bilbao, Spain, for F1_3 and Portugal for C_7. These results show that the HIV-1 epidemic among MSM in Spain is becoming increasingly diverse through the expansion of diverse non-subtype B clusters, comprising or related to viruses circulating in other countries.
RESUMO
HIV-1 protease-reverse transcriptase sequences from 62 HIV-1-infected individuals recently diagnosed in Moscow were analyzed. Subtype A former Soviet Union (FSU) (AFSU) variant was the predominant clade (62.9%), followed by subtype B (22.6%), unique recombinants (6.5%), subtype G (6.5%), and CRF01_AE (1.6%). AFSU predominated among people who inject drugs (88.9%) and heterosexually acquired infections (77.8%), while subtype B was the most prevalent genetic form among men who have sex with men (44%), although AFSU was also frequent in this population (36%). Forty-eight (77.4%) viruses branched within intrasubtype clusters, three of which, of subtype B, had a majority of viruses collected outside of FSU. The four subtype G viruses identified in this study belonged to the Portuguese-Spanish (Iberian) variant and, together with three from databases, formed a Russian cluster closely related to viruses from Denmark. This is the first report of the circulation of the Iberian subtype G variant in Russia.
Assuntos
Variação Genética , Genótipo , Infecções por HIV/virologia , HIV-1/classificação , HIV-1/genética , Análise por Conglomerados , Infecções por HIV/epidemiologia , Protease de HIV/genética , HIV-1/isolamento & purificação , Humanos , Masculino , Epidemiologia Molecular , Moscou/epidemiologia , Filogenia , Análise de Sequência de DNARESUMO
Introducción : El Síndrome Metabólico (SM) es una entidad clínica que se manifiesta por obesidad central, dislipidemia, hiperglicemia y/o hipertensión arterial y se relaciona con el desarrollo de Diabetes Mellitus tipo 2 y enfermedades cardiovasculares. Objetivo : Determinar la prevalencia del SM en adolescentes escolarizados del Departamento Central en el 2021 y evaluar los factores de riesgo relacionados. Materiales y Métodos: Estudio descriptivo con componente analítico. Ingresaron 279 adolescentes de 10 a 16 años de 6 escuelas elegidas aleatoriamente. Se realizó una encuesta sobre estilos de vida, prácticas alimentarias, antecedentes familiares y perinatales. Se realizó medición de peso, talla, circunferencia de cintura (CC), glicemia, perfil lipídico y presión arterial. Para el diagnóstico de SM se utilizó los criterios de Cook. Aprobado por el Comité de Ética del IICS y del Hospital de Clínicas. Para el análisis se utilizaron tablas de frecuencias, mediana y Prueba del Chi cuadrado para razón de prevalencias (RP) con nivel de significancia p p90 36%. La prevalencia de SM fue del 10,7 %. La CC >p90 con RP de 31,4 (7,6-128), y la hipertrigliceridemia con RP de 46,8(14,8-147,7), fueron los componentes preponderantes de SM. El peso elevado al nacer y no realizar actividad física, fueron identificados como factores de riesgo significativo. Conclusiones: La prevalencia de SM fue 10,7%, se relacionó con mayor peso al nacimiento, obesidad y sedentarismo.
Introduction: Metabolic Syndrome (MS) is a clinical entity that is manifested by central obesity, dyslipidemia, hyperglycemia and/or arterial hypertension and is related to the development of Type 2 Diabetes Mellitus and cardiovascular diseases. Objective: To determine the prevalence of MS in adolescents enrolled in school in the Central Department in 2021 and to evaluate their related risk factors. Materials and Methods: This was a descriptive study with an analytical component. 279 adolescents aged 10 to 16 years old from 6 randomly chosen schools were included. A survey on lifestyles, eating practices, family and perinatal history was performed. Weight, height, waist circumference (WC), glycemia, lipid profile, and blood pressure were measured. Cook's criteria were used for the diagnosis of MS. The study was approved by the Ethics Committee of the IICS and the University Hospital. For data analysis, tables of frequencies, median and Chi-square test were used for the prevalence ratio (PR) with a significance level of p p90 in 36%. The prevalence of MS was 10.7%. WC >p90 with a PR of 31.4 (7.6-128), and hypertriglyceridemia with a PR of 46.8 (14.8-147.7), were the predominant components of MS. High birth weight and physical inactivity were identified as significant risk factors. Conclusions: The prevalence of MS was 10.7%, it was related to higher birth weight, obesity and sedentary lifestyle.
RESUMO
In this study, the genetic diversity of HIV-1 in Tunisia was analyzed. For this, 193 samples were collected in different regions of Tunisia between 2012 and 2015. A protease and reverse transcriptase fragment were amplified and sequenced. Phylogenetic analyses were performed through maximum likelihood and recombination was analyzed by bootscanning. Six HIV-1 subtypes (B, A1, G, D, C, and F2), 5 circulating recombinant forms (CRF02_AG, CRF25_cpx, CRF43_02G, CRF06_cpx, and CRF19_cpx), and 11 unique recombinant forms were identified. Subtype B (46.4%) and CRF02_AG (39.4%) were the predominant genetic forms. A group of 44 CRF02_AG sequences formed a distinct Tunisian cluster, which also included four viruses from western Europe. Nine viruses were closely related to isolates collected in other African or in European countries. In conclusion, a high HIV-1 genetic diversity is observed in Tunisia and the local spread of CRF02_AG is first documented in this country.
Assuntos
Variação Genética , Infecções por HIV/virologia , HIV-1/classificação , HIV-1/genética , Análise por Conglomerados , Europa (Continente) , Genótipo , Infecções por HIV/epidemiologia , Protease de HIV/genética , Transcriptase Reversa do HIV/genética , HIV-1/isolamento & purificação , Humanos , Epidemiologia Molecular , Filogenia , Reação em Cadeia da Polimerase , Recombinação Genética , Análise de Sequência de DNA , Tunísia/epidemiologiaRESUMO
BACKGROUND: The incidence of enterovirus D68 (EV-D68) and the spectrum of clinical disease in children are not well known in European countries. We have designed a study with the objective of describing the clinical impact of EV-D68 detected in children with respiratory tract infections. METHODS: As a part of a prospective study to identify the etiology and clinical characteristics of viral respiratory infections in children in Spain, we performed the analysis of the cases of EV infections in all children hospitalized in a secondary hospital in Madrid, during the epidemic respiratory season 2012-2013. A second group of samples was corresponded to infants of the same area, with ambulatory respiratory infection or asymptomatic. Phylogenetic EV-D68 analysis was made using the viral protein 1 gene (VP1). Clinical data of EV-D68 patients were compared with those infected by rhinovirus in the same period and population. RESULTS: The study population consisted of 720 patients corresponding to 399 episodes of hospitalization for respiratory causes, 44 episodes of ambulatory respiratory infections and 277 children determined as a healthy control group. A total of 22 patients were positive for EVs (3.05%), and 12 of them were specifically typed as EV-D68 (11/443 respiratory infections, 2.5%). The most frequent diagnosis in the 10 hospitalized children with EV-D68 detection was recurrent wheezing. Hypoxia was present in 70% of cases, but admission in the intensive care unit was not required. No neurological signs or symptoms were observed. One patient had an ambulatory mild bronchiolitis and another was asymptomatic. No differences were found with rhinovirus infections except less duration of hypoxia and fever in EV-D68 group. CONCLUSIONS: EV-D68 infections were detected in 3.05% of respiratory studied samples (2.5% of admissions). The infection was associated with wheezing episodes with hypoxia. No admissions to intensive care unit or neurological symptoms were found.
Assuntos
Enterovirus Humano D/classificação , Enterovirus Humano D/genética , Infecções por Enterovirus/epidemiologia , Infecções por Enterovirus/virologia , Pacientes Internados , Pacientes Ambulatoriais , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Adolescente , Proteínas do Capsídeo/genética , Criança , Pré-Escolar , Infecções por Enterovirus/diagnóstico , Feminino , Genótipo , Hospitalização , Humanos , Lactente , Recém-Nascido , Masculino , Filogenia , Infecções Respiratórias/diagnóstico , Espanha/epidemiologiaRESUMO
HIV-1 exhibits a characteristically high genetic diversity, with the M group, responsible for the pandemic, being classified into nine subtypes, 72 circulating recombinant forms (CRFs) and numerous unique recombinant forms (URFs). Here we characterize the near full-length genome sequence of an HIV-1 BG intersubtype recombinant virus (X3208) collected in Galicia (Northwest Spain) which exhibits a mosaic structure coincident with that of a previously characterized BG recombinant virus (9601_01), collected in Germany and epidemiologically linked to Portugal, and different from currently defined CRFs. Similar recombination patterns were found in partial genome sequences from three other BG recombinant viruses, one newly derived, from a virus collected in Spain, and two retrieved from databases, collected in France and Portugal, respectively. Breakpoint coincidence and clustering in phylogenetic trees of these epidemiologically-unlinked viruses allow to define a new HIV-1 CRF (CRF73_BG). CRF73_BG shares one breakpoint in the envelope with CRF14_BG, which circulates in Portugal and Spain, and groups with it in a subtype B envelope fragment, but the greatest part of its genome does not appear to derive from CRF14_BG, although both CRFs share as parental strain the subtype G variant circulating in the Iberian Peninsula. Phylogenetic clustering of partial pol and env segments from viruses collected in Portugal and Spain with X3208 and 9691_01 indicates that CRF73_BG is circulating in both countries, with proportions of around 2-3% Portuguese database HIV-1 isolates clustering with CRF73_BG. The fact that an HIV-1 recombinant virus characterized ten years ago as a URF has been shown to represent a CRF suggests that the number of HIV-1 CRFs may be much greater than currently known.
Assuntos
Genoma Viral/genética , HIV-1/genética , Infecções por HIV/virologia , HIV-1/classificação , Humanos , Filogenia , Portugal , EspanhaRESUMO
This work was aimed to study the HIV-1 subtype B epidemics in the Basque Country, Spain. 1727 HIV-1 subtype B sequences comprising protease and reverse transcriptase (PR/RT) coding regions, sampled between 2001 and 2008, were analyzed. 156 transmission clusters were detected by means of phylogenetic analyses. Most of them comprised less than 4 individuals and, in total, they included 441 patients. Six clusters comprised 10 or more patients and were further analyzed in order to study their origin and diversification. Four clusters included men who had unprotected homosexual sex (MSM), one group was formed by intravenous drug users (IDUs), and another included both IDUs and people infected through unprotected heterosexual sex (HTs). Most of these clusters originated from the mid-1980s to the mid-1990s. Only one cluster, formed by MSM, originated after 2000. The time between infections was significantly lower in MSM groups than in those containing IDUs (P-value <0.0001). Nucleoside RT and non-nucleoside RT inhibitor (NRTI and NNRTI)-resistance mutations to antiretroviral treatment were found in these six clusters except the most recent MSM group, but only the IDU clusters presented protease inhibitor (PI)-resistance mutations. The most prevalent mutations for each inhibitor class were PI L90M, NRTI T215D/Y/F, and NNRTI K103N, which were also among the most prevalent resistant variants in the whole dataset. In conclusion, while most infections occur as isolated introductions into the population, the number of infections found to be epidemiologically related within the Basque Country is significant. Public health control measures should be reinforced to prevent the further expansion of transmission clusters and resistant mutations occurring within them.
Assuntos
Farmacorresistência Viral , Infecções por HIV/transmissão , Protease de HIV/genética , Transcriptase Reversa do HIV/genética , HIV-1/classificação , Usuários de Drogas/estatística & dados numéricos , Genótipo , HIV-1/genética , Homossexualidade Masculina/estatística & dados numéricos , Humanos , Masculino , Mutação , Filogenia , Análise de Sequência de RNA/métodos , Espanha/epidemiologia , Fatores de TempoRESUMO
Our objectives were to carry out an epidemiological surveillance study on transmitted drug resistance (TDR) among individuals newly diagnosed of HIV-1 infection during a nine year period in Spain and to assess the role of transmission clusters (TC) in the propagation of resistant strains. An overall of 1614 newly diagnosed individuals were included in the study from January 2004 through December 2012. Individuals come from two different Spanish regions: Galicia and the Basque Country. Resistance mutations to reverse transcriptase inhibitors (RTI) and protease inhibitors (PI) were analyzed according to mutations included in the surveillance drug-resistance mutations list updated in 2009. TC were defined as those comprising viruses from five or more individuals whose sequences clustered in maximum likelihood phylogenetic trees with a bootstrap value ≥90%. The overall prevalence of TDR to any drug was 9.9%: 4.9% to nucleoside RTIs (NRTIs), 3.6% to non-nucleoside RTIs (NNRTIs), and 2.7% to PIs. A significant decrease of TDR to NRTIs over time was observed [from 10% in 2004 to 2% in 2012 (p=0.01)]. Sixty eight (42.2%) of 161 sequences with TDR were included in 25 TC composed of 5 or more individuals. Of them, 9 clusters harbored TDR associated with high level resistance to antiretroviral drugs. T215D revertant mutation was transmitted in a large cluster comprising 25 individuals. The impact of epidemiological networks on TDR frequency may explain its persistence in newly diagnosed individuals. The knowledge of the populations involved in TC would facilitate the design of prevention programs and public health interventions.
Assuntos
Monitoramento Epidemiológico , Infecções por HIV/epidemiologia , Protease de HIV/genética , Transcriptase Reversa do HIV/genética , HIV-1/classificação , HIV-1/genética , Mutação , Adulto , Análise por Conglomerados , Farmacorresistência Viral/efeitos dos fármacos , Farmacorresistência Viral/genética , Feminino , Genótipo , Infecções por HIV/tratamento farmacológico , Infecções por HIV/transmissão , Infecções por HIV/virologia , Protease de HIV/metabolismo , Inibidores da Protease de HIV/uso terapêutico , Transcriptase Reversa do HIV/antagonistas & inibidores , Transcriptase Reversa do HIV/metabolismo , HIV-1/efeitos dos fármacos , HIV-1/enzimologia , Humanos , Masculino , Filogenia , Prevalência , Inibidores da Transcriptase Reversa/uso terapêutico , Espanha/epidemiologiaRESUMO
We recently reported the rapid expansion of an HIV-1 subtype F cluster among men who have sex with men (MSM) in the region of Galicia, Northwest Spain. Here we update this outbreak, analyze near full-length genomes, determine phylogenetic relationships, and estimate its origin. For this study, we used sequences of HIV-1 protease-reverse transcriptase and env V3 region, and for 17 samples, near full-length genome sequences were obtained. Phylogenetic analyses were performed via maximum likelihood. Locations and times of most recent common ancestors were estimated using Bayesian inference. Among samples analyzed by us, 100 HIV-1 F1 subsubtype infections of monophyletic origin were diagnosed in Spain, including 88 in Galicia and 12 in four other regions. Most viruses (n = 90) grouped in a subcluster (Galician subcluster), while 7 from Valladolid (Central Spain) grouped in another subcluster. At least 94 individuals were sexually-infected males and at least 71 were MSM. Seventeen near full-length genomes were uniformly of F1 subsubtype. Through similarity searches and phylogenetic analyses, we identified 18 viruses from four other Western European countries [Switzerland (n = 8), Belgium (n = 5), France (n = 3), and United Kingdom (n = 2)] and one from Brazil, from samples collected in 2005-2011, which branched within the subtype F cluster, outside of both Spanish subclusters, most of them corresponding to recently infected individuals. The most probable geographic origin and age of the Galician subcluster was Ferrol, Northwest Galicia, around 2007, while the Western European cluster probably emerged in Switzerland around 2002. In conclusion, a recently expanded HIV-1 subtype F cluster, the largest non-subtype B cluster reported in Western Europe, continues to spread among MSM in Spain; this cluster is part of a larger cluster with a wide geographic circulation in diverse Western European countries.
Assuntos
Infecções por HIV/epidemiologia , Infecções por HIV/virologia , HIV-1/classificação , HIV-1/genética , Homossexualidade Masculina , Filogenia , Filogeografia , Terapia Antirretroviral de Alta Atividade , Teorema de Bayes , Surtos de Doenças , Farmacorresistência Viral , Europa (Continente)/epidemiologia , Genótipo , Infecções por HIV/tratamento farmacológico , Infecções por HIV/transmissão , HIV-1/efeitos dos fármacos , Humanos , Masculino , Dados de Sequência Molecular , Prevalência , Espanha/epidemiologiaRESUMO
BACKGROUND: HIV-1 subtype B is largely predominant in the Caribbean, although other subtypes have been recently identified in Cuba. OBJECTIVES: To examine HIV-1 genetic diversity in Cuba. METHODS: The study enrolled 105 HIV-1-infected individuals, 93 of whom had acquired the infection in Cuba. DNA from peripheral blood mononuclear cells was used for polymerase chain reaction amplification and sequencing of pol (protease-reverse transcriptase) and env (V3 region) segments. Phylogenetic trees were constructed using the neighbour-joining method. Intersubtype recombination was analysed by bootscanning. RESULTS: Of the samples, 50 (48%) were of subtype B and 55 (52%) of diverse non-B subtypes and recombinant forms. Among non-B viruses, 12 were non-recombinant, belonging to six subtypes (C, D, F1, G, H and J), the most frequent of which was subtype G (n = 5). The remaining 43 (78%) non-B viruses were recombinant, with 14 different forms, the two most common of which were Dpol/Aenv (n = 21) and U(unknown)pol/Henv (n = 7), which grouped in respective monophyletic clusters. Twelve recombinant viruses were mosaics of different genetic forms circulating in Cuba. Overall, 21 genetic forms were identified, with all known HIV-1 group M subtypes present in Cuba, either as non-recombinant viruses or as segments of recombinant forms. Non-B subtype viruses were predominant among heterosexuals (72%) and B subtype viruses among homo- or bisexuals (63%). CONCLUSION: An extraordinarily high diversity of HIV-1 genetic forms, unparalleled in the Americas and comparable to that found in Central Africa, is present in Cuba.
Assuntos
Variação Genética , Infecções por HIV/virologia , HIV-1/genética , Cuba/epidemiologia , Bases de Dados Genéticas , Feminino , Genes pol , Proteína gp120 do Envelope de HIV/genética , Infecções por HIV/epidemiologia , HIV-1/classificação , Humanos , Masculino , Fragmentos de Peptídeos/genética , FilogeniaRESUMO
BACKGROUND: The novel pandemic A (H1N1) pdm09 virus was first identified in Mexico in April 2009 and since then it spread worldwide over a short period of time. Although the virus infection is generally associated with mild disease and a relatively low mortality, it is projected that mutations in specific regions of the viral genome, especially within the receptor binding domain of the haemagglutinin (HA) protein could result in more virulent virus stains, leading to a more severe pathogenicity. METHODS: To monitor the genetic polymorphisms at position 222 of Haemagglutinin of influenza A(H1N1)pdm09 viruses from both outpatients with mild influenza and individuals with severe disease requiring hospitalization, during 2009-2010 and 2010-2011 seasons, a sequence-based genotypic assessment of viral populations to understand the prevalence of D222G mutation. RESULTS: The D222G was identified in clinical specimens from 3 out of 42 cases analyzed in Tunisia with severe outcome (7%). Interestingly, in one fatal case out of four viruses taken from fatal cases studied (25%). Also this mutation was found in one mild case out of 8 mild cases studied (0.1%). D222E substitution was found in virus taken from one patient with severe clinical syndrome (2%) out of 42 severe cases analyzed and E374K substitution was found in two severe cases (4%) out of 42 severe cases studied. CONCLUSIONS: A specific mutation in the viral haemagglutinin (D222G) was found in fatal, severe and mild case. Further virological, clinical and epidemiological investigations are needed to ascertain the role of this and other mutations that may alter the virulence and transmissibility of the pandemic influenza A (H1N1)pdm09. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1027334947811255.
Assuntos
Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Vírus da Influenza A Subtipo H1N1/genética , Influenza Humana/genética , Mutação , Pandemias , Adolescente , Adulto , Feminino , Frequência do Gene , Genótipo , Hospitalização , Humanos , Vírus da Influenza A Subtipo H1N1/patogenicidade , Influenza Humana/diagnóstico , Influenza Humana/epidemiologia , Influenza Humana/transmissão , Influenza Humana/virologia , Masculino , Pessoa de Meia-Idade , Fenótipo , Índice de Gravidade de Doença , Fatores de Tempo , Tunísia/epidemiologia , VirulênciaRESUMO
We analyzed RNA splice site usage in three HIV-1 subtype B primary isolates through reverse transcriptase polymerase chain reaction (RT-PCR) amplification of spliced RNAs using a fluorescently labeled primer, with computerized size determination and quantification of PCR products, which were also identified by clone sequencing. In one isolate, P2149-3, unusual and unreported spliced transcripts were detected. This isolate preferentially used for rev RNA generation a 3' splice site (3'ss) located five nucleotides upstream of A4a, previously identified only in a T cell line-adapted virus and in a group O isolate, and designated A4d. P2149-3 also used an unreported 3'ss for rev RNA generation, designated A4h, located 20 nucleotides upstream of 3'ss A4c. Additionally, unusual nef RNAs using 3'ss A5a and A7a and with exon composition 1.3.7 were identified. The identification of several unusual and unreported spliced transcripts in an HIV-1 primary isolate suggests a greater diversity of splice site usage in HIV-1 than previously appreciated.
Assuntos
Genes nef , Genes rev , HIV-1/genética , Sítios de Splice de RNA/genética , RNA Viral/genética , Sequência de Bases , Variação Genética , HIV-1/classificação , HIV-1/isolamento & purificação , Humanos , Leucócitos Mononucleares/virologia , Dados de Sequência MolecularRESUMO
BACKGROUND: The data contribute to a better understanding of the circulation of influenza viruses especially in North-Africa. OBJECTIVE: The objective of this surveillance was to detect severe influenza cases, identify their epidemiological and virological characteristics and assess their impact on the healthcare system. METHOD: We describe in this report the findings of laboratory-based surveillance of human cases of influenza virus and other respiratory viruses' infection during three seasons in Tunisia. RESULTS: The 2008-09 winter influenza season is underway in Tunisia, with co-circulation of influenza A/H3N2 (56.25%), influenza A(H1N1) (32.5%), and a few sporadic influenza B viruses (11.25%). In 2010-11 season the circulating strains are predominantly the 2009 pandemic influenza A(H1N1)pdm09 (70%) and influenza B viruses (22%). And sporadic viruses were sub-typed as A/H3N2 and unsubtyped influenza A, 5% and 3%, respectively. Unlike other countries, highest prevalence of influenza B virus Yamagata-like lineage has been reported in Tunisia (76%) localised into the clade B/Bangladesh/3333/2007. In the pandemic year, influenza A(H1N1)pdm09 predominated over other influenza viruses (95%). Amino acid changes D222G and D222E were detected in the HA gene of A(H1N1)pdm09 virus in two severe cases, one fatal case and one mild case out of 50 influenza A(H1N1)pdm09 viruses studied. The most frequently reported respiratory virus other than influenza in three seasons was RSV (45.29%). CONCLUSION: This article summarises the surveillance and epidemiology of influenza viruses and other respiratory viruses, showing how rapid improvements in influenza surveillance were feasible by connecting the existing structure in the health care system for patient records to electronic surveillance system for reporting ILI cases.
Assuntos
Influenza Humana/epidemiologia , Orthomyxoviridae/classificação , Vigilância em Saúde Pública , Geografia Médica , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , História do Século XXI , Humanos , Influenza Humana/história , Influenza Humana/virologia , Orthomyxoviridae/genética , Filogenia , Estações do Ano , Vigilância de Evento Sentinela , TunísiaRESUMO
Human Rhinovirus (HRV) classification is an evolving process. New genotypes have been described within HRV-A and HRV-C species, but only one has been accepted related to HRV-B. From 2003 to 2010, a total of 3987 nasopharyngeal aspirate samples were taken from pediatric patients admitted to the Severo Ochoa Hospital in Madrid (Spain). After viral analysis, 949 (23.8%) tested positive to HRV. A random selection of 397 (42%) positive samples showed that 39 (9.8%) were HRV-B. The sequencing of partial VP4/VP2 coding region revealed the spread of 13 of 25 defined HRV-B serotypes and three putative new genotypes. Such results remark the high diversity of HRV-B.
Assuntos
Infecções por Picornaviridae/virologia , Infecções Respiratórias/virologia , Rhinovirus/genética , Rhinovirus/isolamento & purificação , Adolescente , Criança , Criança Hospitalizada , Pré-Escolar , Feminino , Genótipo , Humanos , Lactente , Masculino , Dados de Sequência Molecular , Filogenia , Infecções por Picornaviridae/epidemiologia , Infecções Respiratórias/epidemiologia , Rhinovirus/classificação , EspanhaRESUMO
Pandemic 2009 H1N1 (pH1N1) influenza viruses caused mild symptoms in most infected patients. However, a greater rate of severe disease was observed in healthy young adults and children without co-morbid conditions. Here we tested whether influenza strains displaying differential virulence could be present among circulating pH1N1 viruses. The biological properties and the genotype of viruses isolated from a patient showing mild disease (M) or from a fatal case (F), both without known co-morbid conditions were compared in vitro and in vivo. The F virus presented faster growth kinetics and stronger induction of cytokines than M virus in human alveolar lung epithelial cells. In the murine model in vivo, the F virus showed a stronger morbidity and mortality than M virus. Remarkably, a higher proportion of mice presenting infectious virus in the hearts, was found in F virus-infected animals. Altogether, the data indicate that strains of pH1N1 virus with enhanced pathogenicity circulated during the 2009 pandemic. In addition, examination of chemokine receptor 5 (CCR5) genotype, recently reported as involved in severe influenza virus disease, revealed that the F virus-infected patient was homozygous for the deleted form of CCR5 receptor (CCR5Δ32).