RESUMO
The disease coronavirus COVID-19 has been the cause of millions of deaths worldwide. Among the proteins of SARS-CoV-2, non-structural protein 12 (NSP12) plays a key role during COVID infection and is part of the RNA-dependent RNA polymerase complex. The monitoring of NSP12 polymorphisms is extremely important for the design of new antiviral drugs and monitoring of viral evolution. This study analyzed the NSP12 mutations detected in circulating SARS-CoV-2 during the years 2020 to 2022 in the population of the city of Manaus, Amazonas, Brazil. The most frequent mutations found were P323L and G671S. Reports in the literature indicate that these mutations are related to transmissibility efficiency, which may have contributed to the extremely high numbers of cases in this location. In addition, two mutations described here (E796D and R914K) are close and have RMSD that is similar to the mutations M794V and N911K, which have been described in the literature as influential on the performance of the NSP12 enzyme. These data demonstrate the need to monitor the emergence of new mutations in NSP12 in order to better understand their consequences for the treatments currently used and in the design of new drugs.
Assuntos
COVID-19 , RNA-Polimerase RNA-Dependente de Coronavírus , SARS-CoV-2 , Humanos , Brasil , Simulação por Computador , COVID-19/virologia , COVID-19/transmissão , Mutação/genética , SARS-CoV-2/genética , Proteínas não Estruturais Virais/genética , RNA-Polimerase RNA-Dependente de Coronavírus/genética , RNA-Polimerase RNA-Dependente de Coronavírus/metabolismoRESUMO
We characterized 3 autochthonous dengue virus serotype 3 cases and 1 imported case from 2 states in the North and South Regions of Brazil, 15 years after Brazil's last outbreak involving this serotype. We also identified a new Asian lineage recently introduced into the Americas, raising concerns about future outbreaks.
Assuntos
Aedes , Vírus da Dengue , Dengue , Humanos , Animais , Dengue/epidemiologia , Vírus da Dengue/genética , Sorogrupo , Brasil/epidemiologia , Surtos de DoençasRESUMO
BACKGROUND: Infection with Zika virus (ZIKV) manifests in a broad spectrum of disease ranging from mild illness to severe neurological complications and little is known about Zika immunopathogenesis. OBJECTIVES: To define the immunologic biomarkers that correlate with acute ZIKV infection. METHODS: We characterized the levels of circulating cytokines, chemokines, and growth factors in 54 infected patients of both genders at five different time points after symptom onset using microbeads multiplex immunoassay; comparison to 100 age-matched controls was performed for statistical analysis and data mining. FINDINGS: ZIKV-infected patients present a striking systemic inflammatory response with high levels of pro-inflammatory mediators. Despite the strong inflammatory pattern, IL-1Ra and IL-4 are also induced during the acute infection. Interestingly, the inflammatory cytokines IL-1ß, IL-13, IL-17, TNF-α, and IFN-γ; chemokines CXCL8, CCL2, CCL5; and the growth factor G-CSF, displayed a bimodal distribution accompanying viremia. While this is the first manuscript to document bimodal distributions of viremia in ZIKV infection, this has been documented in other viral infections, with a primary viremia peak during mild systemic disease and a secondary peak associated with distribution of the virus to organs and tissues. MAIN CONCLUSIONS: Biomarker network analysis demonstrated distinct dynamics in concurrence with the bimodal viremia profiles at different time points during ZIKV infection. Such a robust cytokine and chemokine response has been associated with blood-brain barrier permeability and neuroinvasiveness in other flaviviral infections. High-dimensional data analysis further identified CXCL10, a chemokine involved in foetal neuron apoptosis and Guillain-Barré syndrome, as the most promising biomarker of acute ZIKV infection for potential clinical application.
Assuntos
Quimiocinas/imunologia , Citocinas/sangue , Infecção por Zika virus/imunologia , Doença Aguda , Adulto , Biomarcadores/sangue , Estudos de Casos e Controles , Quimiocina CXCL10/sangue , Quimiocinas/sangue , Estudos Transversais , Citocinas/imunologia , Feminino , Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Infecção por Zika virus/sangue , Infecção por Zika virus/complicaçõesRESUMO
The aim of this study was to identify polymorphisms in the folp1, gyrA, and rpoB genes in leprosy patients treated in Amazonas State, Brazil. Among 197 slit-skin smear samples from untreated or relapsed patients, we found three cases of primary resistance to rifampin and one confirmed case of multidrug resistance.
Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Hanseníase/microbiologia , Mycobacterium leprae/efeitos dos fármacos , Mycobacterium leprae/genética , Rifampina/farmacologia , Proteínas de Bactérias/genética , Brasil , Feminino , Humanos , Masculino , Mycobacterium leprae/isolamento & purificação , Polimorfismo GenéticoRESUMO
The SARS-CoV-2 variants of concern (VOCs) Delta and Omicron spread globally during mid and late 2021, respectively. In this study, we compare the dissemination dynamics of these VOCs in the Amazonas state, one of Brazil's most heavily affected regions. We sequenced the virus genome from 4128 patients collected in Amazonas between July 1st, 2021, and January 31st, 2022, and investigated the viral dynamics using a phylodynamic approach. The VOCs Delta and Omicron BA.1 displayed similar patterns of phylogeographic spread but different epidemic dynamics. The replacement of Gamma by Delta was gradual and occurred without an upsurge of COVID-19 cases, while the rise of Omicron BA.1 was extremely fast and fueled a sharp increase in cases. Thus, the dissemination dynamics and population-level impact of new SARS-CoV-2 variants introduced in the Amazonian population after mid-2021, a setting with high levels of acquired immunity, greatly vary according to their viral phenotype.
Assuntos
COVID-19 , SARS-CoV-2 , Humanos , Brasil , Imunidade AdaptativaRESUMO
The rapid spread of the SARS-CoV-2 Variant of Concern (VOC) Gamma in Amazonas during early 2021 fueled a second large COVID-19 epidemic wave and raised concern about the potential role of reinfections. Very few cases of reinfection associated with the VOC Gamma have been reported to date, and their potential impact on clinical, immunological, and virological parameters remains largely unexplored. Here we describe 25 cases of SARS-CoV-2 reinfection in Brazil. SARS-CoV-2 genomic analysis confirmed that individuals were primo-infected with distinct viral lineages between March and December 2020 (B.1.1, B.1.1.28, B.1.1.33, B.1.195, and P.2) and reinfected with the VOC Gamma between 3 to 12 months after primo-infection. We found a similar mean cycle threshold (Ct) value and limited intra-host viral diversity in both primo-infection and reinfection samples. Sera of 14 patients tested 10-75 days after reinfection displayed detectable neutralizing antibodies (NAb) titers against SARS-CoV-2 variants that circulated before (B.1.*), during (Gamma), and after (Delta and Omicron) the second epidemic wave in Brazil. All individuals had milder or no symptoms after reinfection, and none required hospitalization. These findings demonstrate that individuals reinfected with the VOC Gamma may display relatively high RNA viral loads at the upper respiratory tract after reinfection, thus contributing to onward viral transmissions. Despite this, our study points to a low overall risk of severe Gamma reinfections, supporting that the abrupt increase in hospital admissions and deaths observed in Amazonas and other Brazilian states during the Gamma wave was mostly driven by primary infections. Our findings also indicate that most individuals analyzed developed a high anti-SARS-CoV-2 NAb response after reinfection that may provide some protection against reinfection or disease by different SARS-CoV-2 variants.
Assuntos
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , Brasil/epidemiologia , COVID-19/epidemiologia , Diversidade de Anticorpos , Raios gama , Reinfecção , Gravidade do PacienteRESUMO
Dengue virus serotype 2, genotype Cosmopolitan (DENV-2-GII), is one of the most widespread DENV strains globally. In the USA, DENV-2 epidemics have been dominated by DENV-2 genotype Asian-American (DENV-2-GIII), and the first cases of DENV-2-GII were only described in 2019, in Peru, and in 2021 in Brazil. To gain new information about the circulation of DENV-2-GII in Brazil, we sequenced 237 DENV-2 confirmed cases sampled between March 2021 and March 2023 and revealed that DENV-2-GII is already present in all geographic regions of Brazil. The phylogeographic analysis inferred that DENV-2-GII was introduced at least four times in Brazil, between May 2020 and August 2022, generating multiple clades that spread throughout the country with different success. Despite multiple introductions of DENV-2-GII, analysis of the country-wide laboratory surveillance data showed that the Brazilian dengue epidemic in 2022 was dominated by DENV-1 in most states. We hypothesize that massive circulation of DENV-2-GIII in previous years in Brazil might have created a population immune barrier against symptomatic homotypic reinfections by DENV-2-GII, leading to sustained cryptic circulation in asymptomatic cases and localized outbreaks of this new genotype. In summary, our study stresses the importance of arboviral genomic surveillance to close monitoring and better understanding the potential impact of DENV-2-GII in the coming years.
RESUMO
The Amazonas was one of the most heavily affected Brazilian states by the COVID-19 epidemic. Despite a large number of infected people, particularly during the second wave associated with the spread of the Variant of Concern (VOC) Gamma (lineage P.1), SARS-CoV-2 continues to circulate in the Amazonas. To understand how SARS-CoV-2 persisted in a human population with a high immunity barrier, we generated 1,188 SARS-CoV-2 whole-genome sequences from individuals diagnosed in the Amazonas state from 1st January to 6th July 2021, of which 38 were vaccine breakthrough infections. Our study reveals a sharp increase in the relative prevalence of Gamma plus (P.1+) variants, designated Pango Lineages P.1.3 to P.1.6, harboring two types of additional Spike changes: deletions in the N-terminal (NTD) domain (particularly Δ144 or Δ141-144) associated with resistance to anti-NTD neutralizing antibodies or mutations at the S1/S2 junction (N679K or P681H) that probably enhance the binding affinity to the furin cleavage site, as suggested by our molecular dynamics simulations. As lineages P.1.4 (S:N679K) and P.1.6 (S:P681H) expanded (Re > 1) from March to July 2021, the lineage P.1 declined (Re < 1) and the median Ct value of SARS-CoV-2 positive cases in Amazonas significantly decreases. Still, we did not find an increased incidence of P.1+ variants among breakthrough cases of fully vaccinated patients (71%) in comparison to unvaccinated individuals (93%). This evidence supports that the ongoing endemic transmission of SARS-CoV-2 in the Amazonas is driven by the spread of new local Gamma/P.1 sublineages that are more transmissible, although not more efficient to evade vaccine-elicited immunity than the parental VOC. Finally, as SARS-CoV-2 continues to spread in human populations with a declining density of susceptible hosts, the risk of selecting more infectious variants or antibody evasion mutations is expected to increase. IMPORTANCE The continuous evolution of SARS-CoV-2 is an expected phenomenon that will continue to happen due to the high number of cases worldwide. The present study analyzed how a Variant of Concern (VOC) could still circulate in a population hardly affected by two COVID-19 waves and with vaccination in progress. Our results showed that the answer behind that was a new generation of Gamma-like viruses, which emerged locally carrying mutations that made it more transmissible and more capable of spreading, partially evading prior immunity triggered by natural infections or vaccines. With thousands of new cases daily, the current pandemics scenario suggests that SARS-CoV-2 will continue to evolve and efforts to reduce the number of infected subjects, including global equitable access to COVID-19 vaccines, are mandatory. Thus, until the end of pandemics, the SARS-CoV-2 genomic surveillance will be an essential tool to better understand the drivers of the viral evolutionary process.
Assuntos
COVID-19/enzimologia , Furina/metabolismo , SARS-CoV-2/genética , Glicoproteína da Espícula de Coronavírus/química , Glicoproteína da Espícula de Coronavírus/genética , Motivos de Aminoácidos , Brasil/epidemiologia , COVID-19/epidemiologia , COVID-19/transmissão , COVID-19/virologia , Vacinas contra COVID-19/administração & dosagem , Furina/genética , Genômica , Humanos , Mutação , Filogenia , SARS-CoV-2/classificação , SARS-CoV-2/isolamento & purificação , SARS-CoV-2/metabolismo , Glicoproteína da Espícula de Coronavírus/metabolismoRESUMO
The coronavirus disease COVID-19 has been the cause of millions of deaths worldwide. Among the SARS-CoV-2 proteins, the non-structural protein 1 (NSP1) has great importance during the virus infection process and is present in both alpha and beta-CoVs. Therefore, monitoring of NSP1 polymorphisms is crucial in order to understand their role during infection and virus-induced pathogenicity. Herein, we analyzed how mutations detected in the circulating SARS-CoV-2 in the population of the city of Manaus, Amazonas state, Brazil could modify the tertiary structure of the NSP1 protein. Three mutations were detected in the SARS-CoV-2 NSP1 gene: deletion of the amino acids KSF from positions 141 to 143 (delKSF), SARS-CoV-2, lineage B.1.195; and two substitutions, R29H and R43C, SARS-CoV-2 lineage B.1.1.28 and B.1.1.33, respectively. The delKSF was found in 47 samples, whereas R29H and R43C were found in two samples, one for each mutation. The NSP1 structures carrying the mutations R43C and R29H on the N-terminal portion (e.g. residues 10 to 127) showed minor backbone divergence compared to the Wuhan model. However, the NSP1 C-terminal region (residues 145 to 180) was severely affected in the delKSF and R29H mutants. The intermediate variable region (residues 144 to 148) leads to changes in the C-terminal region, particularly in the delKSF structure. New investigations must be carried out to analyze how these changes affect NSP1 activity during the infection. Our results reinforce the need for continuous genomic surveillance of SARS-CoV-2 to better understand virus evolution and assess the potential impact of the viral mutations on the approved vaccines and future therapies.
Assuntos
COVID-19/epidemiologia , SARS-CoV-2/genética , Proteínas não Estruturais Virais/genética , Sequência de Aminoácidos/genética , Substituição de Aminoácidos/genética , Brasil/epidemiologia , Humanos , Polimorfismo Genético/genética , Deleção de Sequência/genéticaRESUMO
The northern state of Amazonas is among the regions in Brazil most heavily affected by the COVID-19 epidemic and has experienced two exponentially growing waves, in early and late 2020. Through a genomic epidemiology study based on 250 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genomes from different Amazonas municipalities sampled between March 2020 and January 2021, we reveal that the first exponential growth phase was driven mostly by the dissemination of lineage B.1.195, which was gradually replaced by lineage B.1.1.28 between May and June 2020. The second wave coincides with the emergence of the variant of concern (VOC) P.1, which evolved from a local B.1.1.28 clade in late November 2020 and replaced the parental lineage in <2 months. Our findings support the conclusion that successive lineage replacements in Amazonas were driven by a complex combination of variable levels of social distancing measures and the emergence of a more transmissible VOC P.1 virus. These data provide insights to understanding the mechanisms underlying the COVID-19 epidemic waves and the risk of dissemination of SARS-CoV-2 VOC P.1 in Brazil and, potentially, worldwide.
Assuntos
COVID-19/epidemiologia , COVID-19/virologia , Genoma Viral/genética , SARS-CoV-2/genética , Adulto , Brasil/epidemiologia , Teste de Ácido Nucleico para COVID-19 , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Filogenia , Filogeografia , Análise Espaço-TemporalRESUMO
A doença do coronavírus COVID-19 tem sido a causa de milhões de mortes em todo o mundo. Dentre as proteínas do SARS-CoV-2, a proteína não estrutural 1 (NSP1) tem grande importância durante o processo de infecção do vírus e está presente tanto em alfa quanto em beta-CoVs. Portanto, o monitoramento dos polimorfismos de NSP1 é crucial para entender seu papel durante a infecção e a patogenicidade induzida pelo vírus. Aqui, analisamos como mutações detectadas no SARS-CoV-2 circulante na população da cidade de Manaus, estado do Amazonas, Brasil, poderiam modificar a estrutura terciária da proteína NSP1. Três mutações foram detectadas no gene SARS-CoV-2 NSP1: deleção dos aminoácidos KSF das posições 141 a 143 (delKSF), SARS-CoV-2, linhagem B.1.195; e duas substituições, R29H e R43C, linhagem SARSCoV-2 B.1.1.28 e B.1.1.33, respectivamente. O delKSF foi encontrado em 47 amostras, enquanto R29H e R43C foram encontrados em duas amostras, uma para cada mutação. As estruturas NSP1 que carregam as mutações R43C e R29H na porção N-terminal (por exemplo, resíduos 10 a 127) mostraram divergência de espinha dorsal menor em comparação com o modelo Wuhan. No entanto, a região C-terminal de NSP1 (resíduos 145 a 180) foi severamente afetada nos mutantes delKSF e R29H. A região variável intermediária (resíduos 144 a 148) leva a alterações na região C-terminal, particularmente na estrutura delKSF. Novas investigações devem ser realizadas para analisar como essas alterações afetam a atividade da NSP1 durante a infecção. Nossos resultados reforçam a necessidade de vigilância genômica contínua do SARS-CoV-2 para entender melhor a evolução do vírus e avaliar o impacto potencial das mutações virais nas vacinas aprovadas e futuras terapias.
RESUMO
Aponta as causas do crescimento do número de casos de Covid-19 no Amazonas e das sucessivas substituições de linhagens do Sars-CoV-2. O estudo indicou que esses fatores foram impulsionados por uma combinação de diminuições das medidas de distanciamento social e pelo surgimento de uma forma mais transmissível do vírus, a variante P.1, identificada em meados de novembro de 2020. Essa variante causou um aumento exponencial da doença, o que estabeleceu a segunda onda da epidemia no estado.
Assuntos
Infecções por Coronavirus , Coronavirus , SARS-CoV-2 , Ecossistema Amazônico , BrasilRESUMO
OBJECTIVES: To determine the etiology and factors associated with genital ulcer disease (GUD) among patients presenting to a sexually transmitted infections clinic in Manaus, Brazil; and to compare a multiplex polymerase chain reaction (M-PCR) assay for the diagnosis of GUD with standard methods. METHODS: Ulcer swabs were collected and used for Tzanck test and processed in an M-PCR to detect herpes simplex virus (HSV-1/2), Treponema pallidum (T. pallidum), and Haemophilus ducreyi (H. ducreyi). Sera were tested for HIV and syphilis antibodies. Multivariable analysis was used to measure the association between clinical aspects and GUD. M-PCR results were compared with syphilis serology and Tzanck tests. RESULTS: Overall, 434 GUD samples were evaluated, 84.8% from men. DNA from HSV-2 was detected in 55.3% of GUD samples, T. pallidum in 8.3%, HSV-1 in 3.2%, and 32.5% of GUD specimens were negative for the DNA of all three pathogens. No cases of H. ducreyi were identified. HIV serology among GUD patients was 3.2%. Treponemal antibodies and Tzanck test positivity for genital herpes was detected in 25 (5.8%) and in 125 (30.3%) of GUD patients, respectively. In multivariable analysis genital herpes etiology by M-PCR was associated with the vesicular, multiple and recurrent lesions whereas T. pallidum with non-vesicular, non-recurrent lesions. Compared to M-PCR, syphilis serology was 27.8% sensitive and 96.2% specific whereas Tzanck test was 43.8% sensitive and 88.9% specific. CONCLUSIONS: The predominance of genital herpes etiology suggests a revision of existing national syndromic treatment guidelines in Brazil to include antiherpetic treatment for all GUD patients. The use of M-PCR can significantly improve the diagnosis of GUD and provide a greater sensitivity than standard diagnostics.
Assuntos
Infecções Sexualmente Transmissíveis/microbiologia , Infecções Sexualmente Transmissíveis/virologia , Adulto , Brasil/epidemiologia , Demografia , Feminino , Humanos , Masculino , Reação em Cadeia da Polimerase Multiplex , Prevalência , Fatores de Risco , Infecções Sexualmente Transmissíveis/sangue , Infecções Sexualmente Transmissíveis/epidemiologia , Simplexvirus/genética , Sífilis/sangue , Sífilis/microbiologia , Sífilis/virologia , Treponema pallidum/genética , Adulto JovemRESUMO
BACKGROUND Infection with Zika virus (ZIKV) manifests in a broad spectrum of disease ranging from mild illness to severe neurological complications and little is known about Zika immunopathogenesis. OBJECTIVES To define the immunologic biomarkers that correlate with acute ZIKV infection. METHODS We characterized the levels of circulating cytokines, chemokines, and growth factors in 54 infected patients of both genders at five different time points after symptom onset using microbeads multiplex immunoassay; comparison to 100 age-matched controls was performed for statistical analysis and data mining. FINDINGS ZIKV-infected patients present a striking systemic inflammatory response with high levels of pro-inflammatory mediators. Despite the strong inflammatory pattern, IL-1Ra and IL-4 are also induced during the acute infection. Interestingly, the inflammatory cytokines IL-1β, IL-13, IL-17, TNF-α, and IFN-γ; chemokines CXCL8, CCL2, CCL5; and the growth factor G-CSF, displayed a bimodal distribution accompanying viremia. While this is the first manuscript to document bimodal distributions of viremia in ZIKV infection, this has been documented in other viral infections, with a primary viremia peak during mild systemic disease and a secondary peak associated with distribution of the virus to organs and tissues. MAIN CONCLUSIONS Biomarker network analysis demonstrated distinct dynamics in concurrence with the bimodal viremia profiles at different time points during ZIKV infection. Such a robust cytokine and chemokine response has been associated with blood-brain barrier permeability and neuroinvasiveness in other flaviviral infections. High-dimensional data analysis further identified CXCL10, a chemokine involved in foetal neuron apoptosis and Guillain-Barré syndrome, as the most promising biomarker of acute ZIKV infection for potential clinical application.
Assuntos
Humanos , Quimiocina CXCL10/sangue , Infecção por Zika virus/complicações , Expressão Gênica , Quimiocinas/imunologia , Infecção por Zika virus/imunologiaRESUMO
Dentre as 27 Unidades Federativas brasileiras, o estado do Amazonas foi uma das mais afetadas pela presente pandemia do novo coronavírus, com a ocorrência de uma segunda onda ao final de 2020 na qual um grande número de casos graves levou a um colapso no sistema de saúde. Este pre-print, resultado da análise do genoma completo de 250 amostras do SARS-CoV-2 coletadas no Amazonas entre março de 2020 e janeiro de 2021, descreve a dinâmica de sucessões de linhagens dominantes no estado. Mais recentemente, ele foi publicado na revista Nature Medicine, após editoração e a revisão por pesquisadores independentes. A publicação apresenta dados que embasam as hipóteses de que a linhagem responsável pela maioria dos casos no primeiro momento da pandemia (entre março e maio de 2020) foi a B.1.195, de que esta foi suplantada pela B.1.1.28, que tornou-se a linhagem dominante no estado entre maio e dezembro de 2020, e de que em dezembro o surgimento de uma variante da B.1.1.28, denominada P.1 e dotada de maior transmissibilidade, foi responsável pela nova ascensão no número de casos e mortes. Desta forma, a dinâmica local de surgimento de novas genéticas virais foi uma importante força-motriz para a forma com a qual a pandemia avançou sobre o estado do Amazonas, influenciada diretamente pela circulação da população e sua relação com o espalhamento do vírus, que culminou com a substituição da B.1.1.28 pela variante de preocupação P.1 em um processo que acredita-se ter durado apenas dois meses.
RESUMO
Neste relatório (pre-print), são apresentados três casos de reinfecção causados pela Variante de Preocupação (VOC) P.1, também conhecida como "cepa de Manaus". As três pacientes eram mulheres adultas, e tiveram a primeira infecção durante a primeira onda da pandemia na primeira metade de 2020. Nos três casos, a linhagem detectada no primeiro diagnóstico molecular era diferente da encontrada posteriormente, evidência da reinfecção. Dois dos casos de reinfecção tiveram apresentação de sintomas leves, enquanto o terceiro foi assintomático, apesar de a quantidade de material genético viral detectado sugerir cargas virais elevadas. As evidências aqui apresentadas sugerem que a imunidade após infecção primária por linhagens anteriores à circulação daquelas contendo a mutação E484K não impede uma nova infecção pela variante P.1, e nem mesmo que pessoas reinfectadas por esta variante espalhem o vírus, embora seja possível que tenha protegido estas três pacientes do desenvolvimento de sintomas graves.