CAMILA MALTA ROMANO
Projetos de Pesquisa
Unidades Organizacionais
LIM/52 - Laboratório de Virologia, Hospital das Clínicas, Faculdade de Medicina
7 resultados
Resultados de Busca
Agora exibindo 1 - 7 de 7
- Glytube: A Conical Tube and Parafilm M-Based Method as a Simplified Device to Artificially Blood-Feed the Dengue Vector Mosquito, Aedes aegypti(2013) COSTA-DA-SILVA, Andre Luis; NAVARRETE, Flavia Rosa; SALVADOR, Felipe Scassi; KARINA-COSTA, Maria; IOSHINO, Rafaella Sayuri; AZEVEDO, Diego Soares; ROCHA, Desiree Rafaela; ROMANO, Camila Malta; CAPURRO, Margareth LaraAedes aegypti, the main vector of dengue virus, requires a blood meal to produce eggs. Although live animals are still the main blood source for laboratory colonies, many artificial feeders are available. These feeders are also the best method for experimental oral infection of Ae. aegypti with Dengue viruses. However, most of them are expensive or laborious to construct. Based on principle of Rutledge-type feeder, a conventional conical tube, glycerol and Parafilm-M were used to develop a simple in-house feeder device. The blood feeding efficiency of this apparatus was compared to a live blood source, mice, and no significant differences (p = 0.1189) were observed between artificial-fed (51.3% of engorgement) and mice-fed groups (40.6%). Thus, an easy to assemble and cost-effective artificial feeder, designated ""Glytube"" was developed in this report. This simple and efficient feeding device can be built with common laboratory materials for research on Ae. aegypti.
- Inter- and Intra-Host Viral Diversity in a Large Seasonal DENV2 Outbreak(2013) ROMANO, Camila Malta; LAUCK, Michael; SALVADOR, Felipe S.; LIMA, Celia Rodrigues; VILLAS-BOAS, Lucy S.; ARAUJO, Evaldo Stanislau A.; LEVI, Jose Eduardo; PANNUTI, Claudio Sergio; O'CONNOR, David; KALLAS, Esper GeorgesBackground: High genetic diversity at both inter-and intra-host level are hallmarks of RNA viruses due to the error-prone nature of their genome replication. Several groups have evaluated the extent of viral variability using different RNA virus deep sequencing methods. Although much of this effort has been dedicated to pathogens that cause chronic infections in humans, few studies investigated arthropod-borne, acute viral infections. Methods and Principal Findings: We deep sequenced the complete genome of ten DENV2 isolates from representative classical and severe cases sampled in a large outbreak in Brazil using two different approaches. Analysis of the consensus genomes confirmed the larger extent of the 2010 epidemic in comparison to a previous epidemic caused by the same viruses in another city two years before (genetic distance = 0.002 and 0.0008 respectively). Analysis of viral populations within the host revealed a high level of conservation. After excluding homopolymer regions of 454/Roche generated sequences, we found 10 to 44 variable sites per genome population at a frequency of >1%, resulting in very low intra-host genetic diversity. While up to 60% of all variable sites at intra-host level were non-synonymous changes, only 10% of inter-host variability resulted from non-synonymous mutations, indicative of purifying selection at the population level. Conclusions and Significance: Despite the error-prone nature of RNA-dependent RNA-polymerase, dengue viruses maintain low levels of intra-host variability.
- An international, interlaboratory ring trial confirms the feasibility of an extraction-less ""direct"" RT-qPCR method for reliable detection of SARS-CoV-2 RNA in clinical samples(2022) MILLS, Margaret G.; BRUCE, Emily; HUANG, Meei-Li; CROTHERS, Jessica W.; HYRIEN, Ollivier; OURA, Christopher A. L.; BLAKE, Lemar; JORDAN, Arianne Brown; HESTER, Susan; WEHMAS, Leah; MARI, Bernard; BARBY, Pascal; LACOUX, Caroline; FASSY, Julien; VIAL, Pablo; VIAL, Cecilia; MARTINEZ, Jose R. W.; OLADIPO, Olusola Olalekan; INUWA, Bitrus; SHITTU, Ismaila; MESEKO, Clement A.; CHAMMAS, Roger; SANTOS, Carlos Ferreira; DIONISIO, Thiago Jose; GARBIERI, Thais Francini; PARISI, Viviane Aparecida; MENDES-CORREA, Maria Cassia; PAULA, Anderson V. de; ROMANO, Camila M.; GOES, Luiz Gustavo Bentim; MINOPRIO, Paola; CAMPOS, Angelica C.; CUNHA, Marielton P.; VILELA, Ana Paula P.; NYIRENDA, Tonney; MKAKOSYA, Rajhab Sawasawa; MUULA, Adamson S.; DUMM, Rebekah E.; HARRIS, Rebecca M.; MITCHELL, Constance A.; PETTIT, Syril; BOTTEN, Jason; JEROME, Keith R.Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is used worldwide to test and trace the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). ""Extraction-less"" or ""direct"" real time-reverse transcription polymerase chain reaction (RT-PCR) is a transparent and accessible qualitative method for SARS-CoV-2 detection from nasopharyngeal or oral pharyngeal samples with the potential to generate actionable data more quickly, at a lower cost, and with fewer experimental resources than full RT-qPCR. This study engaged 10 global testing sites, including laboratories currently experiencing testing limitations due to reagent or equipment shortages, in an international interlaboratory ring trial. Participating laboratories were provided a common protocol, common reagents, aliquots of identical pooled clinical samples, and purified nucleic acids and used their existing in-house equipment. We observed 100% concordance across laboratories in the correct identification of all positive and negative samples, with highly similar cycle threshold values. The test also performed well when applied to locally collected patient nasopharyngeal samples, provided the viral transport media did not contain charcoal or guanidine, both of which appeared to potently inhibit the RT-PCR reaction. Our results suggest that direct RT-PCR assay methods can be clearly translated across sites utilizing readily available equipment and expertise and are thus a feasible option for more efficient COVID-19 coronavirus disease testing as demanded by the continuing pandemic.
- Torquetenovirus in saliva: A potential biomarker for SARS-CoV-2 infection?(2021) MENDES-CORREA, Maria C.; TOZETTO-MENDOZA, Tania Regina; FREIRE, Wilton S.; PAIAO, Heuder G. O.; FERRAZ, Andrea B. C.; MAMANA, Ana C.; FERREIRA, Noely E.; V, Anderson de Paula; FELIX, Alvina C.; ROMANO, Camila M.; BRAZ-SILVA, Paulo H.; LEAL, Fabio E.; GRESPAN, Regina M. Z.; SABINO, Ester C.; COSTA, Silvia F.; WITKIN, Steven S.Torquetenovirus (TTV) is present in biological fluids from healthy individuals and measurement of its titer is used to assess immune status in individuals with chronic infections and after transplants. We assessed if the titer of TTV in saliva varied with the presence of SARS-CoV-2 in the nasopharynx and could be a marker of COVID-19 status. Saliva from 91 individuals positive for SARS-CoV-2 in nasal-oropharyngeal samples, and from 126 individuals who were SARS-CoV-2-negative, all with mild respiratory symptoms, were analyzed. Both groups were similar in age, gender, symptom duration and time after symptom initiation when saliva was collected. Titers of TTV and SARS-CoV-2 were assessed by gene amplification. Loss of smell (p = 0.0001) and fever (p = 0.0186) were more prevalent in SARS-CoV-2-positive individuals, while sore throat (p = 0.0001), fatigue (p = 0.0037) and diarrhea (p = 0.0475) were more frequent in the SARS-CoV-2 negative group. The saliva TTV and nasal-oropharyngeal SARS-CoV-2 titers were correlated (p = 0.0085). The TTV level decreased as symptoms resolved in the SARS-CoV-2 infected group (p = 0.0285) but remained unchanged in the SARS-CoV-2 negative controls. In SARS-CoV-2 positive subjects who provided 2-4 saliva samples and in which TTV was initially present, the TTV titer always decreased over time as symptoms resolved. We propose that sequential TTV measurement in saliva is potentially useful to assess the likelihood of symptom resolution in SARS-CoV-2-positive individuals and to predict prognosis.
- First report of Aedes albopictus infected by Dengue and Zika virus in a rural outbreak in Brazil(2020) REZENDE, Helder Ricas; ROMANO, Camila Malta; CLARO, Ingra Morales; CALEIRO, Giovana Santos; SABINO, Ester Cerdeira; FELIX, Alvina Clara; BISSOLI, Jefferson; HILL, Sarah; FARIA, Nuno Rodrigues; SILVA, Theresa Cristina Cardoso da; SANTOS, Ana Paula Brioschi; CERUTTI JUNIOR, Crispim; VICENTE, Creuza RachelIn Brazil, Dengue (DENV) and Zika (ZIKV) viruses are reported as being transmitted exclusively by Aedes aegypti in urban settings. This study established the vectors and viruses involved in an arbovirus outbreak that occurred in 2019 in a rural area of Espi ' rito Santo state, Brazil. Mosquitoes collected were morphologically identified, sorted in samples, and submitted to molecular analysis for arboviruses detection. Phylogenetic reconstruction was performed for the viral sequence obtained. All 393 mosquitoes were identified as Aedes albopictus. DENV-1 genotype V was present in one sample and another sample was positive for ZIKV. The DENV-1 clustered with viruses that have circulated in previous years in large urban centers of different regions in Brazil. This is the first report of A. albopictus infected by DENV and ZIKV during an outbreak in a rural area in Brazil, indicating its involvement in arboviral transmission. The DENV-1 strain found in the A. albopictus was not new in Brazil, being involved previously in epidemics related to A. aegypti, suggesting the potential to A. albopictus in transmitting viruses already circulating in the Brazilian population. This finding also indicates the possibility of these viruses to disperse across urban and rural settings, imposing additional challenges for the control of the diseases.
- Serum from dengue virus-infected patients with and without plasma leakage differentially affects endothelial cells barrier function in vitro(2017) CARDOZO, Francielle Tramontini Gomes de Sousa; BAIMUKANOVA, Gyulnar; LANTERI, Marion Christine; KEATING, Sheila Marie; FERREIRA, Frederico Moraes; HEITMAN, John; PANNUTI, Claudio Sergio; PATI, Shibani; ROMANO, Camila Malta; SABINO, Ester CerdeiraBackground Although most of cases of dengue infections are asymptomatic or mild symptomatic some individuals present warning signs progressing to severe dengue in which plasma leakage is a hallmark. Methodology/Principal findings The present study used Electric Cell-substrate Impedance Sensing (ECIS (R)) which allows for electrical monitoring of cellular barrier function measuring changes in Transendothelial Electric Resistance (TEER) to investigate the parameters associated with dengue induced leakage. Three groups of individuals were tested: dengue-positives with plasma leakage (leakage), dengue-positives without plasma leakage (no leakage), and dengue-negatives (control). Data show that TEER values of human umbilical vein endothelial cells (HUVECs) was significantly lower after incubation with serum from subjects of the leakage group in comparison to the no leakage or control groups. The serum levels of CXCL1, EGF, eotaxin, IFN-gamma, sCD40L, and platelets were significantly decreased in the leakage group, while IL-10, IL-6, and IP-10 levels were significantly increased. We also found a strong correlation between TEER values and augmented levels of IP-10, GM-CSF, IL-1 alpha, and IL-8, as well as decreased levels of CXCL1 and platelets. Conclusions/Significance The present work shows that the magnitude of the immune response contributes to the adverse plasma leakage outcomes in patients and that serum components are important mediators of changes in endothelial homeostasis during dengue infections. In particular, the increased levels of IP-10 and the decreased levels of CXCL1 and platelets seem to play a significant role in the disruption of vascular endothelium associated with leakage outcomes after DENV infection. These findings may have important implications for both diagnostic and therapeutic approaches to predict and mitigate vascular permeabilization in those experiencing the most severe clinical disease outcomes after dengue infection.
- Determination of clusters and factors associated with dengue dispersion during the first epidemic related to Dengue virus serotype 4 in Vitoria, Brazil(2017) VICENTE, Creuza Rachel; HERBINGER, Karl-Heinz; CERUTTI JUNIOR, Crispim; ROMANO, Camila Malta; CABIDELLE, Aline de Souza Areias; FROESCHL, GuenterDengue occurrence is partially influenced by the immune status of the population. Consequently, the introduction of a new Dengue virus serotype can trigger explosive epidemics in susceptible populations. The determination of clusters in this scenario can help to identify hotspots and understand the disease dispersion regardless of the influence of the population herd immunity. The present study evaluated the pattern and factors associated with dengue dispersion during the first epidemic related to Dengue virus serotype 4 in Vitoria, Espirito Santo state, Brazil. Data on 18,861 dengue cases reported in Vitoria from September 2012 to June 2013 were included in the study. The analysis of spatial variation in temporal trend was performed to detect clusters that were compared by their respective relative risk, house index, population density, and income in an ecological study. Overall, 11 clusters were detected. The time trend increase of dengue incidence in the overall study population was 636%. The five clusters that showed a lower time trend increase than the overall population presented a higher incidence in the beginning of the epidemic and, compared to the six clusters with higher time trend increase, they presented higher relative risk for their inhabitants to acquire dengue infection (P-value = 0.02) and a lower income (P-value < 0.01). House index and population density did not differ between the clusters. Early increase of dengue incidence and higher relative risk for acquiring dengue infection were favored in low-income areas. Preventive actions and improvement of infrastructure in low-income areas should be prioritized in order to diminish the magnitude of dengue dispersion after the introduction of a new serotype.