Isolation and characterization of a Brazilian strain of yellow fever virus from an epizootic outbreak in 2009

Jorge, Taissa Ricciardi; Mosimann, Ana Luiza Pamplona; Noronha, Lúcia de; Maron, Angela; Santos, Claudia Nunes Duarte dos

doi:10.1016/j.actatropica.2016.09.030

Cited by 8 publications

(6 citation statements)

References 31 publications

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“…YFV RNA investigation was performed by real-time PCR preceded by reverse transcription (RT-qPCR), using GoTaq Probe 1-Step RT-qPCR System (Promega) and primers and probe described by Domingo and colleagues (2012) [ 14 ]. Using specific primers, targeting CprM and envelope regions of the YFV genome [ 15 ], partial sequences were amplified and sequenced by dideoxy-method on an ABI3130 platform (Applied Biosystems). Raw data were analyzed and final contigs were assembled using SeqTrace [ 16 ].…”

Section: Methodsmentioning

confidence: 99%

Persistence of Yellow fever virus outside the Amazon Basin, causing epidemics in Southeast Brazil, from 2016 to 2018

et al. 2018

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BackgroundYellow fever (YF) is endemic in the Brazilian Amazon Basin, and sporadic outbreaks take place outside the endemic area in Brazil. Since 2016, YF epidemics have been occurring in Southeast Brazil, with more than 1,900 human cases and more than 1,600 epizooties of non-human primates (NHPs) reported until April 2018. Previous studies have demonstrated that Yellow fever virus (YFV) causing outbreaks in 2017 formed a monophyletic group.Methodology/Principal findingsAiming to decipher the origin of the YFV responsible for the recent epidemics, we obtained nucleotide sequences of YFV detected in humans (n = 6) and NHPs (n = 10) from Minas Gerais state during 2017–2018. Next, we performed evolutionary analyses and discussed the results in the light of epidemiological records (official numbers of YFV cases at each Brazilian Federative unit, reported by the Brazilian Ministry of Health). Nucleotide sequences of YFV from Southeast Brazil from 2016 to 2018 were highly conserved and formed a monophyletic lineage (BR-YFV_2016/18) within the genotype South America I. Different clusters were observed within lineage BR-YFV_2016/18, one containing the majority of isolates (from humans and NHPs), indicating the sylvatic transmission of YFV. We also detected a cluster characterized by two synapomorphies (amino acid substitutions) that contained YFV only associated with NHP what should be further investigated. The topology of lineage BR-YFV_2016/18 was congruent with epidemiological and temporal patterns of the ongoing epidemic. YFV isolates detected in 2016, in São Paulo state were located in the most basal position of the lineage, followed by the isolates from Minas Gerais and Espírito Santo obtained in 2017 and 2018. The most recent common ancestor of the lineage BR-YFV_2016/18 dated to 2015 (95% credible intervals = 2014–2016), in a period that was coincident with the reemergence of YFV in the Midwest region of Brazil.ConclusionsThe results demonstrated a single introduction of YFV in the Southeast region and the silent viral circulation before the onset of the outbreaks in 2016. Evolutionary analyses combined with epidemiological records supported the idea that BR-YFV_2016/18 was probably introduced from the Midwest into the Southeast region, possibly in São Paulo state. The persistence of YFV in the Southeast region, causing epidemics from 2016 to 2018, suggests that this region presents suitable ecological and climatic conditions for YFV maintenance during the epidemic and interepidemic seasons. This fact poses risks for the establishing of YF enzootic cycles and epidemics, outside the Amazon Basin in Brazil. YF surveillance and studies of viral dynamics deserve particular attention, especially in Midwest, Southeast and neighbor regions which are the main areas historically associated with YF outbreaks outside the Amazon Basin. YFV persistence in Southeast Brazil should be carefully considered in the context of public health, especially for public health decision-makers and researchers.

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Section: Methodsmentioning

confidence: 99%

Persistence of Yellow fever virus outside the Amazon Basin, causing epidemics in Southeast Brazil, from 2016 to 2018

et al. 2018

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“…The YFV-17DD vaccine strain stock was obtained from two passages of the commercial vaccine (Biomanguinhos, Rio de Janeiro, Brazil) in Huh7.5 cells. The M17/09 YF wild strain 29 corresponds to the second passage in C6/36 cells after virus isolation. Saint Louis encephalitis virus (SLEV) and WNV were propagated in VERO-E6 cells, whereas DENV-1 and DENV-3 were propagated in C6/36 cells.…”

Section: Methodsmentioning

confidence: 99%

Development of a quantitative NS1-capture enzyme-linked immunosorbent assay for early detection of yellow fever virus infection

Jorge

Bordignon

Koishi

et al. 2017

Sci Rep

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Yellow fever is an arboviral disease that causes thousands of deaths every year in Africa and the Americas. However, few commercial diagnostic kits are available. Non-structural protein 1 (NS1) is an early marker of several flavivirus infections and is widely used to diagnose dengue virus (DENV) infection. Nonetheless, little is known about the dynamics of Yellow fever virus (YFV) NS1 expression and secretion, to encourage its use in diagnosis. To tackle this issue, we developed a quantitative NS1-capture ELISA specific for YFV using a monoclonal antibody and recombinant NS1 protein. This test was used to quantify NS1 in mosquito and human cell line cultures infected with vaccine and wild YFV strains. Our results showed that NS1 was detectable in the culture supernatants of both cell lines; however, a higher concentration was maintained as cell-associated rather than secreted into the extracellular milieu. A panel of 73 human samples was used to demonstrate the suitability of YFV NS1 as a diagnostic tool, resulting in 80% sensitivity, 100% specificity, a 100% positive predictive value and a 95.5% negative predictive value compared with RT-PCR. Overall, the developed NS1-capture ELISA showed potential as a promising assay for the detection of early YF infection.

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“…Formalin-fixed paraffin-embedded (FF-PE) brain samples were stained using a conventional haematoxylin-eosin (H&E) staining technique [13] and analyzed by photomicrographs acquired using a Zeiss Axio Scan.Z1 scanner (Carl Zeiss, US). This procedure was performed in the Experimental Pathology Laboratory of Universidade Pontifícia Catolica in Curitiba, Brazil.…”

Section: Histologymentioning

confidence: 99%

Inactivated rabies vaccines: Standardization of an in vitro assay for residual viable virus detection

et al. 2020

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Human rabies, a neglected viral zoonosis, is preventable through domestic animals vaccination and post-exposure prophylaxis using inactivated rabies vaccines. During vaccine production, several mandatory in vivo quality control trials, such as potency, live virus, and safety, are responsible for the use of large numbers of laboratory animals. Over the years, global organizations encouraged the development of alternative methods to reduce, replace and refine the use of animals in the pharmaceutical industry. In this study we standardized an in vitro assay for determination of residual live virus combining viral isolation techniques with direct immunofluorescence detection and viral quantification by a molecular method. Standardization of viral recovery steps and quantification by RT-qPCR were performed and the combined method was shown to be 3 fold more sensitive than the in vivo assay. It was possible to identify viral suspensions cultures, which still had residual viable rabies virus particles, evidencing the importance to implement this method in quality control schemes of rabies vaccine production. In addition, this developed assay is more practical, inexpensive and less time consuming, producing results in just 4 days, which may allow greater agility in the internal quality control of the vaccine. The in vitro method may reduce 2/3 rd of laboratory animals numbers used for this purpose, since it can be applied in the intermediate quality control of inactivated rabies vaccine production.

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Isolation and characterization of a Brazilian strain of yellow fever virus from an epizootic outbreak in 2009

Cited by 8 publications

References 31 publications

Persistence of Yellow fever virus outside the Amazon Basin, causing epidemics in Southeast Brazil, from 2016 to 2018

Persistence of Yellow fever virus outside the Amazon Basin, causing epidemics in Southeast Brazil, from 2016 to 2018

Development of a quantitative NS1-capture enzyme-linked immunosorbent assay for early detection of yellow fever virus infection

Inactivated rabies vaccines: Standardization of an in vitro assay for residual viable virus detection

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