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Comparative studies of third heterologous doses following the CoronaVac vaccine against coronavirus disease 2019 (COVID-19) in kidney transplant recipients are lacking.This prospective, single-center cohort study included kidney transplant recipients without previous COVID-19. Patients received a third heterologous (BNT162b2 mRNA) or homologous dose at least 4 wk after 2 doses of the CoronaVac vaccine. Immunoglobulin G antibody response and seroprevalence for neutralizing anti-severe acute respiratory syndrome coronavirus 2 antibodies immediately before and 28 d after third doses were compared between the groups.There were 307 patients in the heterologous group and 777 in the homologous group. Patients in the heterologous group were older (54 versus 50 y; P0.0001), with a longer time since transplant (11 versus 6 y; P0.0001). Immediately before the third dose, immunoglobulin G seroprevalence (36% versus 34%; P = 0.597) and antibody titers (246 versus 268 AU/mL; P = 0.279) were similar. After booster, seroconversion was higher in the heterologous group (49% versus 32%; P0.0001), resulting in a higher seroprevalence (67% versus 55%; P = 0.0003); however, 42% of all patients remained seronegative. Antibody titers after booster in seropositive patients were higher in the heterologous group (7771 versus 599 AU/mL; P0.0001). These results persisted after adjusting for confounding variables. Lastly, a similar proportion of patients became seropositive for neutralizing antibodies (98% versus 94%; P = 0.098).In kidney transplant recipients fully vaccinated with CoronaVac, a third dose with an mRNA vaccine produced a higher seroconversion rate and antibody titers than a third homologous dose. However, both boosters achieved equivalent seroprevalence for neutralizing antibodies. The high proportion of still seronegative patients indicates the need for alternative strategies of protection.

Monkeypox disease is a viral zoonosis with symptoms similar to those seen in the past in smallpox (variola), although clinically less severe. Following the eradication of smallpox in 1980 and the subsequent cessation of smallpox vaccination, monkeypox has emerged as the most important orthopoxvirus from a public health standpoint. Monkeypox virus occurs primarily in central and western Africa, often in tropical forests, and has increasingly manifested in urban areas. Animal hosts include various rodents and nonhuman primates. We report the case of a patient with monkeypox disease who developed ocular complaints (eye discomfort and conjunctivitis) and had detectable conjunctival lesions on biomicroscopy and fluorescein testing. Its ophthalmological manifestations are still poorly known. RESUMO Varíola do Macaco é uma zoonose viral com sintomas semelhantes aos observados no passado em pacientes com Varíola, embora seja clinicamente menos grave. Com a erradicação da varíola em 1980 e a subsequente cessação da vacinação contra a varíola, a varíola dos macacos emergiu como o ortopoxvírus mais importante em saúde pública. O vírus monkeypox ocorre principalmente na África central e ocidental, muitas vezes nas proximidades de florestas tropicais, e tem se manifestado cada vez mais em áreas urbanas. Os hospedeiros animais incluem uma variedade de roedores e primatas não humanos. O presente estudo relata o caso de um paciente com Monkeypox que evoluiu com queixa oftalmológica de desconforto ocular e conjuntivite e, à biomicroscopia e teste da fluoresceína, detecção de lesões conjuntivais. Alterações oftalmológicas da doença são, ainda, pouco conhecidas.

Objective: To describe the first COVID-19 pandemic at Casa Ondina Lobo, a philanthropic nursing home in São Paulo city, and the containment measures against the pandemic that proved to be effective. Methods: Several preventive measures were taken before and during the pandemic, with emphasis on universal testing by reverse transcription polymerase chain reaction for COVID-19. All residents and employees were tested twice in a D9 period. Results: Among the 62 residents and 55 employees, in both testing, eight residents and nine employees tested positive for COVID-19. Of 22% of employees and 75% of residents evolved asymptomatic, emphasizing the importance of universal testing for the detection and isolation of these cases. A quarter of residents evolved without any symptoms, however, with COVID-19 signs, reinforcing the importance of monitoring vital signs. The second testing did not detect any new cases among residents, demonstrating the effectiveness of the containment measures, however, it found four new cases among employees. This emphasized their role in COVID-19 outbreaks in nursing homes. Only one patient died, a 12.5% lethality among those known to be infected and a 1.6% mortality in the total population of residents were seen. Conclusion: The adoption of appropriate containment measures enabled to contain an COVID-19 pandemic in studied nursing home. Universal reverse transcription polymerase chain reaction testing for COVID-19 has proved to be particularly important and effective.

Em função da velocidade com que a COVID-19 se expandiu, e da extensão com que atingiu a população brasileira, foi necessário um esforço sem precedentes dos laboratórios clínicos para oferecer testes diagnósticos adequados. O conhecimento dos métodos laboratoriais disponíveis para o diagnóstico da infecção e pós-infecção pelo SARS-CoV-2 é fundamental para o manejo da pandemia. Dessa forma, essa revisão tem por objetivo apresentar uma atualização sobre os principais testes diagnósticos usados no momento, seus princípios bioquímicos, aplicações e limitações. Tradicionalmente pouco utilizados para investigação de infecções de trato respiratório, os testes sorológicos são hoje amplamente empregados para o diagnóstico de COVID-19; com custo inferior (comparado aos testes moleculares) e relativa rapidez para liberação de resultado. A sorologia é um exame importante para apoio à decisão diagnóstica, principalmente nos pacientes em período pós-convalescência, e num contexto epidemiológico, para avaliação da soroprevalência na população. Aspectos como a dinâmica da produção dos anticorpos e seu papel na imunização contra a COVID-19 ainda carecem de investigação para que haja maior compreensão e interpretação dos dados. O diagnóstico padrão é feito por métodos moleculares, principalmente por PCR (reação em cadeia da polimerase) em tempo real. A presença do vírus é avaliada pela existência do RNA viral na amostra respiratória; portanto, duas grandes limitações do teste molecular são a qualidade da coleta e a conservação da região genômica alvo do ensaio molecular. Apesar de serem comparativamente mais caros, os testes baseados em PCR são os que apresentam maiores sensibilidade e especificidade no diagnóstico da COVID-19; melhorias processuais e validação de reagentes alternativos foram incorporadas ao fluxo do exame, viabilizando o oferecimento. Uma nova abordagem para o diagnóstico da infecção pelo vírus SARS-CoV-2 por meio da detecção de proteínas virais por proteômica direcionada baseada em espectrometria de massas foi recentemente descrita por nosso grupo. Apesar de não alcançar a sensibilidade do teste de PCR, visto que as proteínas não podem ser multiplicadas como os ácidos nucleicos, o novo teste facilita a logística de coleta e transporte das amostras. Foi verificado que as proteínas são mais estáveis, permitindo o diagnóstico mesmo após o armazenamento das amostras em temperatura ambiente, possibilitando assim o envio de amostras de locais remotos. Unitermos: SARS-CoV-2. COVID-19. Diagnóstico laboratorial. Testes sorológicos. Diagnóstico molecular.

Hepatitis E virus (HEV) affects 20 million people worldwide, with 3.3 million cases and 56,000 deaths. The transmission is mainly by the fecal-oral route. Several studies have reported increased alanine aminotransferase (ALT) levels in association with viral hepatitis. This study evaluated the diagnosis of HEV infection among patients attending the emergency room (ER) of Hospital Beneficencia Portuguesa (HBP) and Hospital Sao Paulo (HSP) in Sao Paulo, Brazil increased ALT levels (≥ 200 IU/L). From October 2018 to July 2019, 400 sera samples were collected from patients treated at the ER of HBP (n=200) and HSP (n=200). All samples were screened for HEV by RT-qPCR. 200 samples from HSP were tested for IgM of anti-Hepatitis A (HAV) and B (HBV) viruses, and total antibodies of Hepatitis C virus (HCV). Ninety samples (45 from each hospital), were tested for anti-HEV IgM antibodies. Patients aged under 1 to 91 years (mean = 46.29 ± 24.17, median = 48). ALT levels varied from 200 to 8,974 IU/l. 16 patients (4%) turned out positive for HEV by RT-qPCR (ALT levels = 299 to 698 IU/L). Of the 200 HSP patients, 18 (9%) were anti-HAV IgM reactive, 9 (4.5%) for anti-HBV IgM, and 7 (3.5%) for anti-HCV antibodies (ALT levels = 833 to 1918 IU/L). Two of 90 BPH patients (2.22%) were anti-HEV IgM reactive (ALT levels = 1502 to 3831 IU/L). This is the first Brazilian study evaluating patients with suspected HEV infection with increased ALT levels, which were higher than 12 and 60 times the normal upper limit, in the acute phase or for patients reactive for antibody detection, respectively. Liver damage could be minimized by implementing molecular diagnostic tests in the hospital routine.

Made available in DSpace on 2022-04-28T19:43:29Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Background: Kidney transplant recipients have higher COVID-19 associated mortality compared to the general population. However, as only symptomatic patients seek medical attention, the current level of exposure, the main sources of acquisition, and the behavior of humoral immunity over time are poorly understood. Methods: This cross-sectional prospective single-center study recruited kidney transplant recipients of any age living in Sao Paulo. A sample size of 401 patients was calculated considering the 17.2% seroprevalence in the municipality population from a published survey, a 95% confidence interval and an absolute error of 2%. Results: Of the 2636 eligible patients, 416 were included. The seroprevalence for IgG anti-SARS-CoV-2 was 8.2%. Seroconversion rate decreased with increasing age, from 15.7% (18–35 years) to 8.3% (36–60 years) and 4.2% (>60 years, p = 0.042). Seropositivity among previously confirmed COVID-19 patients was 68.4%, followed by 9.4% in those with flu-like symptoms and only 4.6% among asymptomatic patients (p

Background The Complete Blood Count (CBC) is a commonly used low-cost test that measures white blood cells, red blood cells, and platelets in a person’s blood. It is a useful tool to support medical decisions, as intrinsic variations of each analyte bring relevant insights regarding potential diseases. In this study, we aimed at developing machine learning models for COVID-19 diagnosis through CBCs, unlocking the predictive power of non-linear relationships between multiple blood analytes. Methods We collected 809,254 CBCs and 1,088,385 RT-PCR tests for SARS-Cov-2, of which 21% (234,466) were positive, from 900,220 unique individuals. To properly screen COVID-19, we also collected 120,807 CBCs of 16,940 individuals who tested positive for other respiratory viruses. We proposed an ensemble procedure that combines machine learning models for different respiratory infections and analyzed the results in both the first and second waves of COVID-19 cases in Brazil. Results We obtain a high-performance AUROC of 90 + % for validations in both scenarios. We show that models built solely of SARS-Cov-2 data are biased, performing poorly in the presence of infections due to other RNA respiratory viruses. Conclusions We demonstrate the potential of a novel machine learning approach for COVID-19 diagnosis based on a CBC and show that aggregating information about other respiratory diseases was essential to guarantee robustness in the results. Given its versatile nature, low cost, and speed, we believe that our tool can be particularly useful in a variety of scenarios—both during the pandemic and after.

Poliomyelitis is still an endemic disease in Afghanistan, Nigeria, and Pakistan despite the efforts to eradicate the disease. Therefore, there is a potential risk of international spread. Since the start of the polio eradication program by the Global Polio Eradication Initiative in 1988, the incidence of polio has been reduced by 99%. In the last decade, wild poliovirus type 2 (WPV2) was eliminated and declared eradicated in 2015. Wild poliovirus type 3 (WPV3) was last reported in November 2012. These changes have allowed the removal of Sabin poliovirus type 2 from the oral poliovirus vaccine (OPV) in April 2016 and countries either introduced bivalent OPV (bOPV) containing Sabin types 1 + 3 poliovirus or added at least one dose of inactivated poliovirus vaccine (IPV) into their routine immunization schedule. Many efforts are needed to eradicate polio, and new strategies should be implemented such as the development and approval of new genetically stable OPV, and vaccines that do not require infectious processes for virus growth, such as virus-like particles (VLPs), or packing-cell technology. IPV will increasingly be produced from Sabin strains, and further attenuated or genetically modified strains. Furthermore, there is also a need for the development of antiviral drugs to treat immunodeficient patients who are long-term excretors infected with poliovirus, thus avoiding contamination of individuals susceptible to polioviruses, due to reversal of pathogenicity. If all these measures are successfully implemented, the world will be close to the global RESUMEN La poliomielitis sigue siendo una enfermedad endémica en Afganistán, Nigeria y Pakistán a pesar de los esfuerzos por erradicar la enfermedad. Por lo tanto, existe un riesgo de propagación mundial. Desde el inicio del programa de erradicación de la poliomielitis por la Iniciativa de Erradicación Mundial de la Poliomielitis [Global Polio Eradication Initiative (GPEI)] en 1988, la incidencia de la poliomielitis se ha reducido en un 99%. En la última década, el poliovirus salvaje tipo 2 (WPV2) fue eliminado y declarado erradicado en 2015. El poliovirus salvaje tipo 3 (WPV3) se informó por última vez en noviembre de 2012. Estos cambios han permitido la eliminación del poliovirus Sabin tipo 2 de la vacuna antipoliomielítica oral (VPO) en abril de 2016, y los países introdujeron la VPO de tipo bivalente (bVPO), que contiene poliovirus Sabin tipos 1 y 3, o agregaron al menos una dosis de vacuna antipoliomielítica inactivada (VPI) al programa de inmunización de rutina. Se necesitan muchos esfuerzos para erradicar la poliomielitis y se deben implementar nuevas estrategias, como el desarrollo y aprobación de nuevas VPO genéticamente estables y vacunas que no requieren procesos infecciosos para el crecimiento del virus, como partículas pseudovirales (VLP) o tecnología de células empaquetadas (packing-cell). La VIP se producirá cada vez más a partir de cepas Sabin y otras cepas más atenuadas o modificadas genéticamente. Además, también es necesario desarrollar fármacos antivirales para tratar a pacientes inmunodeficientes que son excretores a largo plazo, evitando así la contaminación de individuos susceptibles a poliovirus, debido a la reversión de la patogenicidad. Si todas estas medidas se implementan con éxito, el mundo estará cerca de la interrupción global de la transmisión del WPV y la erradicación de la poliomielitis. RESUMO A poliomielite ainda é uma doença endêmica no Afeganistão, na Nigéria e no Paquistão, apesar dos esforços para erradicá-la. Portanto, há risco de propagação mundial. Desde o início do programa de erradicação da poliomielite pela Iniciativa de Erradicação Global da Pólio [Global Polio Eradication Initiative (GPEI)], em 1988, a incidência da doença foi reduzida em 99%. Na última década, o poliovírus selvagem do tipo 2 (WPV2) foi eliminado e declarado erradicado em 2015. O poliovírus selvagem do tipo 3 (WPV3) foi reportado pela última vez em novembro de 2012. Essas mudanças promoveram a remoção do poliovírus Sabin tipo 2 da vacina oral antipólio (VOP) em abril de 2016, e os países introduziram a vacina oral bivalente (VOPb), que contém os poliovírus Sabin tipos 1 + 3, ou adicionaram pelo menos uma dose da vacina inativada contra o poliovírus (VIP) no calendário de imunização. É necessário muito empenho para erradicar a poliomielite. Novas estratégias devem ser implementadas, como o desenvolvimento e a aprovação de novas VOPs geneticamente estáveis e vacinas que não requerem processos infecciosos para o crescimento do vírus, como partículas pseudovirais (VLP), ou tecnologia de células de empacotamento (packing-cell). A VIP será cada vez mais produzida a partir de cepas Sabin, de outras cepas atenuadas ou geneticamente modificadas. Além disso, é imprescindível o desenvolvimento de medicamentos antivirais para tratar os pacientes imunodeficientes que são excretores de longo prazo, evitando assim a contaminação de indivíduos suscetíveis aos poliovírus, devido à reversão da patogenicidade. Se todas essas medidas forem implementadas com sucesso, o mundo estará próximo da interrupção global de transmissão do WPV e da erradicação da poliomielite.

In recent years the immunomodulatory actions of vitamin D, a steroid hormone, have been extensively studied. In 2020, due to the COVID-19 pandemic, the question arose as to 25(OH)D status would be related to susceptibility to SARS-CoV-2 infection, since several studies pointed out a higher prevalence and severity of the disease in populations with low levels of 25(OH)D. Thus, we investigated the 25(OH)D levels in adults “Detected” positive for SARS CoV-2 by RT-PCR (reverse transcriptase polymerase chain reaction) test, and in negative controls, “not Detected”, using the Fleury Group's examination database, in Sao Paulo, Brazil. Of a total of 14.692 people with recent assessments of 25(OH)D and RT-PCR tests for COVID-19, 2.345 were positive and 11.585 were negative for the infection. The groups did not differ in the percentage of men and women, or in the age distribution. There were no differences in the distribution of 25(OH)D between the two groups (p = 0.08); mean 25(OH)D of 28.8 ± 21.4 ng/mL and 29.6 ± 18.1 ng/mL, respectively. In the specific population studied, clinical, environmental, socioeconomic and cultural factors should have greater relevance than 25(OH)D in determining the susceptibility to COVID-19.

The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is pressing public health systems around the world, and large population testing is a key step to control this pandemic disease. Here, we develop a high-throughput targeted proteomics assay to detect SARS-CoV-2 nucleoprotein peptides directly from nasopharyngeal and oropharyngeal swabs. A modified magnetic particle-based proteomics approach implemented on a robotic liquid handler enables fully automated preparation of 96 samples within 4 hours. A TFC-MS system allows multiplexed analysis of 4 samples within 10 min, enabling the processing of more than 500 samples per day. We validate this method qualitatively (Tier 3) and quantitatively (Tier 1) using 985 specimens previously analyzed by real-time RT-PCR, and detect up to 84% of the positive cases with up to 97% specificity. The presented strategy has high sample stability and should be considered as an option for SARS-CoV-2 testing in large populations., Large population testing is a key step to controlling the COVID-19 pandemic. Here, the authors develop a targeted mass spectrometry system for peptide-based SARS-CoV-2 detection, allowing analysis of over 500 swab samples per day and enabling virus detection even after prolonged sample storage at room temperature.

The current outbreak of severe acute respiratory syndrome associated with coronavirus 2 (SARS-CoV-2) is pressing public health systems around the world, and large population testing is a key step to control this pandemic disease. Real-time reverse-transcription PCR (real-time RT-PCR) is the gold standard test for virus detection but the soaring demand for this test resulted in shortage of reagents and instruments, severely limiting its applicability to large-scale screening. To be used either as an alternative, or as a complement, to real-time RT-PCR testing, we developed a high-throughput targeted proteomics assay to detect SARS-CoV-2 proteins directly from clinical respiratory tract samples. Sample preparation was fully automated by using a modified magnetic particle-based proteomics approach implemented on a robotic liquid handler, enabling a fast processing of samples. The use of turbulent flow chromatography included four times multiplexed on-line sample cleanup and UPLC separation. MS/MS detection of three peptides from SARS-CoV-2 nucleoprotein and a 15N-labeled internal global standard was achieved within 2.5 min, enabling the analysis of more than 500 samples per day. The method was validated using 562 specimens previously analyzed by real-time RT-PCR and was able to detect over 83% of positive cases. No interference was found with samples from common respiratory viruses, including other coronaviruses (NL63, OC43, HKU1, and 229E). The strategy here presented has high sample stability and low cost and should be considered as an option to large population testing.

A population-based household survey was performed to estimate the prevalence of IgM and IgG to SARS-CoV-2 in residents of six districts in São Paulo City, Brazil. Serum samples collected from 299 randomly-selected adults and 218 cohabitants (N=517) were tested by chemiluminescence immunoassay ten weeks after the first reported case. Weighted overall seroprevalence was 4.7% (95% CI 3.0-6.6%). The low seroprevalence suggests that most of this population could still be infected. Serial serosurveys were initiated aiming to monitor the progress of the ongoing pandemic throughout the entire city. This may help inform public health authority decisions regarding prevention and control strategies.

Brazil currently has one of the fastest growing SARS-CoV-2 epidemics in the world. Due to limited available data, assessments of the impact of non-pharmaceutical interventions (NPIs) on virus transmission and epidemic spread remain challenging. We investigate the impact of NPIs in Brazil using epidemiological, mobility and genomic data. Mobility-driven transmission models for São Paulo and Rio de Janeiro cities show that the reproduction number (Rt) reached below 1 following NPIs but slowly increased to values between 1 to 1.3 (1.0–1.6). Genome sequencing of 427 new genomes and analysis of a geographically representative genomic dataset from 21 of the 27 Brazilian states identified >100 international introductions of SARS-CoV-2 in Brazil. We estimate that three clades introduced from Europe emerged between 22 and 27 February 2020, and were already well-established before the implementation of NPIs and travel bans. During this first phase of the epidemic establishment of SARS-CoV-2 in Brazil, we find that the virus spread mostly locally and within-state borders. Despite sharp decreases in national air travel during this period, we detected a 25% increase in the average distance travelled by air passengers during this time period. This coincided with the spread of SARS-CoV-2 from large urban centers to the rest of the country. In conclusion, our results shed light on the role of large and highly connected populated centres in the rapid ignition and establishment of SARS-CoV-2, and provide evidence that current interventions remain insufficient to keep virus transmission under control in Brazil.One Sentence SummaryJoint analysis of genomic, mobility and epidemiological novel data provide unique insight into the spread and transmission of the rapidly evolving epidemic of SARS-CoV-2 in Brazil.

In 2007, the new polyomaviruses WUPyV and KIPyV were identified in patients with acute respiratory infections. The aim of this study was to investigate these viruses in hospitalized patients with severe acute respiratory infection (SARI). A retrospective study was conducted with 251 patients, from April 2009 to November 2010, using nasopharyngeal aspirates, naso- and oropharyngeal swab samples from hospitalized patients (children 38.8 °C), dyspnea, and cough. Clinical and epidemiological information was obtained through standardized questionnaire. Enrolled patients were initially suspected to have influenza A(H1N1)pdm09 infections. WUPyV and KIPyV were detected by real-time PCR. Samples were also tested for influenza A and B viruses, human respiratory syncytial virus, rhinovirus, metapneumovirus, coronavirus, adenovirus, and parainfluenza viruses. WUPyV and KIPyV were detected in 6.77% (4.78% and 1.99%, respectively) of hospitalized patients with SARI. All samples from children showed coinfections (rhinovirus was the most commonly detected). Six adults had polyomavirus infection and four (1.6%) had monoinfection. Of them, 3 reported comorbidities including immunosuppression and 1 patient had worse outcome, requiring ICU admission. These preliminary data may suggest a possible role of polyomaviruses in SARI among immunocompromised adult patients.

Background The respiratory syncytial virus (RSV) is recognized as an important cause of respiratory tract infections. Immunocompromised patients, healthcare workers (HCWs) and children contacts are at increased risk of acquiring the infection. However, the impact of asymptomatic infection in transmission has not been well studied. Objectives: this study evaluated the frequency and viral load (VL) of RSV in nasal swab samples of individuals with different risk factors for acquiring infection in a university hospital in Sao Paulo, Brazil. Methods We included 196 symptomatic children and their 192 asymptomatic caregivers, 70 symptomatic and 95 asymptomatic HCWs, 43 samples from symptomatic HIV‐positive outpatients, and 100 samples of asymptomatic HIV patients in the period of 2009‐2013. Results RSV infection was detected in 10.1% (70/696) of samples, 4.4% (17/387) of asymptomatic patients, and 17.1% (53/309) from symptomatic patients. (P < .0001). The VL of symptomatic patients (4.7 log copies/mL) was significantly higher compared to asymptomatic patients (2.3 log copies/mL). RSV detection among asymptomatic caregivers (6.8%; 13/192) was significantly higher compared to other asymptomatic adults, HIV and HCWs (2.0%; 4/195; P = .0252). A close contact with an infected child at home was an important risk to RSV acquisition [OR 22.6 (95% CI 4.8‐106.7)]. Children who possibly transmitted the virus to their asymptomatic contacts had significantly higher viral load than children who probably did not transmit (P < .0001). Conclusions According to our results, it is important to know if people circulating inside the hospital have close contact with acute respiratory infected children.

While the new Coronavirus Disease 2019 (COVID-19) pandemic rapidly spread across the world, South America was reached later in relation to Asia, Europe and the United States of America (USA). Brazil concentrates now the largest number of cases in the continent and, as the disease speedily progressed throughout the country, prompt and challenging operational strategies had to be taken by institutions caring for COVID-19 and non-COVID-19 patients in order to assure optimal workflows, triage, and management. Although hospitals in the USA, Europe and Asia have shared their experience on this subject, little has been discussed about such strategies in South America or by the perspective of outpatient centers, which are paramount in the radiology field. This article shares the guidelines adopted early in the pandemic by a nationwide outpatient healthcare center composed by a network of more than 200 patient service centers and nearly 2,000 radiologists in Brazil, discussing operational and patient management strategies, staff protection, changes adopted in the fellowship program, and the effectiveness of such measures.

Diagnosis of primary immunodeficiency diseases (PID), based on laboratory tests and assessment of T lymphocyte function, is crucial in patients who present with lymphopenia. We evaluated T lymphocyte function in healthy children and adults and in patients and with PID using flow cytometry. Whole blood cultures were stimulated with phytohemagglutinin, purified protein derivate (PPD) and candidin, followed by detection of intracellular interferon-gamma (IFN - gamma) and CD25 membrane expression on CD3+ T cells by flow cytometry. Flow cytometry results were compared with 3H-thymidine (3HTdR) lymproliferation after in vitro cell stimulation and with delayed type hypersensitivity reaction (DTH). Patients with PID had lower intracellular IFN - gamma production than healthy children and healthy adults after PHA stimulation for 18 h ( p = 0.024 and p p = 0.048 and p p p = 0.029). The DTH for PPD was not different between PID and healthy children ( p = 0.281). Intracellular IFN-gamma after PHA stimulation for 18 h and CD25 membrane expression after candidin stimulation for 72 h on CD3+ T cells were most reliable parameters that could discriminate PID patients from healthy children. Our results confirm the high variability in functional cell assays and reinforce the idea that age differences must be taken into consideration during assay evaluation.

Breast cancer remains a leading cause of morbidity and mortality worldwide yet methods for early detection remain elusive. We describe the discovery and validation of biochemical signatures measured by mass spectrometry, performed upon blood samples from patients and controls that accurately identify (>95%) the presence of clinical breast cancer. Targeted quantitative MS/MS conducted upon 1225 individuals, including patients with breast and other cancers, normal controls as well as individuals with a variety of metabolic disorders provide a biochemical phenotype that accurately identifies the presence of breast cancer and predicts response and survival following the administration of neoadjuvant chemotherapy. The metabolic changes identified are consistent with inborn-like errors of metabolism and define a continuum from normal controls to elevated risk to invasive breast cancer. Similar results were observed in other adenocarcinomas but were not found in squamous cell cancers or hematologic neoplasms. The findings describe a new early detection platform for breast cancer and support a role for pre-existing, inborn-like errors of metabolism in the process of breast carcinogenesis that may also extend to other glandular malignancies. Statement of Significance: Findings provide a powerful tool for early detection and the assessment of prognosis in breast cancer and define a novel concept of breast carcinogenesis that characterizes malignant transformation as the clinical manifestation of underlying metabolic insufficiencies.

Poliovirus has been nearly eliminated as part of a world-wide effort to immunize and contain circulating wild-type polio. Nevertheless, poliovirus has been detected in water supplies and represents a threat to patients with humoral immunodeficiencies where infection can be fatal. To define the risk, we analyzed antibodies to poliovirus 1, 2, and 3 in serum samples collected over a year from patients with primary immunodeficiency diseases (PID) on regular intravenous immunoglobulin (IVIG) replacement. Twenty-one patients on regular IVIG replacement therapy were evaluated: Twelve patients with common variable immune deficiency (CVID), six with X-linked agammaglobulinemia (XLA), and three with hyper IgM syndrome (HIGM). Over 1 year, four blood samples were collected from each of these patients immediately before immunoglobulin infusion. One sample of IVIG administered to each patient in the month before blood collection was also evaluated. Poliovirus antibodies were quantified by seroneutralization assay. All IVIG samples had detectable antibodies to the three poliovirus serotypes. Despite that, only 52.4, 61.9, and 19.0% of patients showed protective antibody titers for poliovirus 1, 2, and 3, respectively. Only two patients (9.5%) had protective antibodies for the three poliovirus serotypes on all samples. Most patients were therefore susceptible to all three poliovirus serotypes. This study demonstrates the need for ongoing vigilance regarding exposure of patients with PID to poliovirus in the community.

Brazil has been classified as moderately endemic for hepatitis E virus (HEV) infection. However, data on the seroprevalence of HEV in this region are limited. This study evaluated the prevalence of past or present HEV infection among blood donors in the metropolitan area of Itajai Valley, Southern Brazil, a region of predominant German heritage, where cultural habits result in a high consumption of pork. Serum samples from 300 blood donors were tested in December 2014 using serological and molecular methods. Anti-HEV IgG antibodies were detected in 30 (10%) subjects, and categorized age groups revealed an age-dependent increase of HEV seroprevalence. Only one subject had anti-HEV IgM, whereas none tested positive for HEV-RNA. The present data demonstrate a higher seroprevalence of anti-HEV IgG in blood donors than previously reported in Brazil. J. Med. Virol. 9999: 1–4, 2015. © 2015 Wiley Periodicals, Inc.

In 2009, Influenza A(H1N1)pdm09 had a major effect on global health, causing thousands of deaths. However, the results of asymptomatic infection in transmission has not been well studied. We analyzed asymptomatic influenza infection in individuals with different risk factors for acquiring influenza in a university hospital.Respiratory samples from 100 children with respiratory symptoms, their asymptomatic caregivers, 100 asymptomatic HIV-infected patients, and 100 health care workers collected during 2009-2011 were tested for influenza by real time real-time polymerase chain reaction.Influenza infection rate in symptomatic children was 44% and in asymptomatic adults it was 8.3% (P .01). Children older than age 5 years had a 3.4 times greater chance of being infected during influenza season than younger children. Among the asymptomatic group, influenza was more frequent in caregivers (14%; P = .032) and a higher rate (31.8%) was observed if a child was infected during the 2009 pandemic. Contact with an infected child was an important risk factor for influenza acquisition (odds ratio, 3.8; 95% confidence interval, 1.1-13.2) among caregivers. The mean cluster of differentiation 4 T-cell count of HIV-positive patients infected with influenza was not statistically different from uninfected patients (P = .29).Asymptomatic household contacts of symptomatic children may play an important role in community transmission and a more proactive intervention should be considered during future pandemics.

Background & Aims: Hepatitis C (HC) is a major cause of hepatocellular carcinoma (HCC), and a late diagnosis is the main factor for the poor survival of patients. There is an urgent need for identifying sensitive and specific biomarkers for HCC diagnosis. In the present study, plasma lipid patterns of patients with HC-HCC, HC-liver cirrhosis (LC), and chronic HC (CHC) were assessed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS).Methods. Plasma samples of 25 patients with HC-HCC, 15 patients with HC-LC, and 25 patients with CHC were evaluated by MALDI-MS using a Q-ToF premier (Synapt) mass spectrometer (Waters, Manchester, UK) equipped with a 200-Hz solid-state laser in the mass range between m/z (mass-to-charge ratio) of 700-1200.Results. A total of 2205 ions were initially obtained and 7 ions (m/z) were highlighted as corresponding to the most important lipids to differentiate HCC patients from LC and CHC patients. The specific lipidomic expression signature generated resulted in an overall predictive accuracy of 93% of HC-HCC and HC-LC, and 100% of HC-HCC and CHC. The 7-peak algorithm distinguished HCC from LC with a sensitivity of 96% and a specificity of 87%, and HCC from CHC with both sensitivity and specificity of 100%.Conclusion. MALDI-MS-specific signature peaks accurately distinguished patients with HC-HCC from those with HC-LC and CHC. The results indicate the potential of MALDI-MS and the selected peaks to improve HCC surveillance in patients with viral C cirrhosis and chronic hepatitis C.

Human herpesvirus 6 (HHV-6) may cause severe complications after haematopoietic stem cell transplantation (HSCT). Monitoring this virus and providing precise, rapid and early diagnosis of related clinical diseases, constitute essential measures to improve outcomes. A prospective survey on the incidence and clinical features of HHV-6 infections after HSCT has not yet been conducted in Brazilian patients and the impact of this infection on HSCT outcome remains unclear. A rapid test based on real-time quantitative polymerase chain reaction (qPCR) has been optimised to screen and quantify clinical samples for HHV-6. The detection step was based on reaction with TaqMan® hydrolysis probes. A set of previously described primers and probes have been tested to evaluate efficiency, sensitivity and reproducibility. The target efficiency range was 91.4% with linearity ranging from 10-106 copies/reaction and a limit of detection of five copies/reaction or 250 copies/mL of plasma. The qPCR assay developed in the present study was simple, rapid and sensitive, allowing the detection of a wide range of HHV-6 loads. In conclusion, this test may be useful as a practical tool to help elucidate the clinical relevance of HHV-6 infection and reactivation in different scenarios and to determine the need for surveillance.

Rotavirus is the main global cause of severe childhood diarrhoea among children. In 2006, Rotarix® (G1P[8]) was introduced into Brazil’s National Immunization Program. The vaccine coverage rate was 84.4% in 2009. Evidences of increasing G2P[4] after 2006 opened up the discussion about the vaccine effectiveness to non-G1 strains. The aim of this study was to identify the circulating rotavirus genotypes in two Brazilian regions during 2009. A total of 223 positive samples by immunochromatography and latex agglutination assay from the Northeast (Bahia/Pernambuco States) and Southeast (São Paulo/Rio de Janeiro States) regions were included in the study. The samples were submitted to genotyping by nested-PCR according to VP7(G) and VP4(P) and 175 samples (78.5%) were able to be characterized. Considering the characterization of VP7, the G-types detected were G1, G2, and G4 in the Northeast, and G2, G3, G5, and G9 in the Southeast. Considering the characterization of VP4, the P-types detected were P[4], P[8], and P[6]/P[9] in the Northeast and the Southeast. The most frequent mixed types found were G2P[4]/G2P[NT](81.4%), G2P[6](5.2%), G1P[6](5.2%) in the Northeast, and G2P[4]/G2P[NT](78.8%), G2P[6](8.2%), G9P[8](4.7%) in the Southeast. Among immunized individuals whose age ranged from 0-4 years, the G2P[4]/G2P[NT] genotype was identified in 91,0% of cases, and among non-immunized individuals of the same age, the G2P[4]/G2P[NT] genotype was identified in 85.7% of the cases. In accordance with the high level of vaccine coverage, the data suggest that the circulation of G2P[4] in these regions had a considerable increase after the introduction of Rotarix®.

Objective: Bacterial keratitis occurs worldwide, and despite recent developments, it remains a potentially blinding condition. This study assesses the presence of herpes simplex virus (HSV-1 and -2) and varicella zoster virus (VZV) by quantitative real-time polymerase chain reaction (qPCR) in corneal scrapings from patients with bacterial keratitis. Methods: A total of 65 patients with clinical diagnoses of infectious corneal ulcers prospectively underwent clinical eye examinations. Corneal scrapings were investigated by Gram staining, Giemsa staining, culture, and qPCR (the study group). Risk factors and epidemiological data were recorded. The control group comprising 25 eyes with typical herpes dendritic keratitis was also analyzed by qPCR. Results: From the study group (n=65), nine patients (13.8%) had negative smears, cultures, and qPCR findings. Fifty-six (86.2%) patients had positive cultures: 51 for bacteria, 4 for fungi, and 1 for amoebae. Of the patients who had positive bacterial cultures, qPCR identified 10 patients who were also positive for virus: one for VZV and nine for HSV-1. Of the 25 patients in the control group, 21 tested positive for HSV-1 by qPCR analysis. Conclusions: Herpes may be present in patients with bacterial corneal ulcers, and qPCR may be useful in its detection. RESUMO Objetivo: Ceratites bacterianas ocorrem mundialmente e apesar dos novos desenvolvimentos permanece como uma condição que pode levar à cegueira. Avaliar a presença de herpes simples (-1 e -2) e vírus varicella zoster (VZV) por reação em cadeia quantitativa de polimerase em tempo real (qPCR) em raspados corneanos de pacientes com ceratite bacteriana. Métodos: Sessenta e cinco pacientes com ceratite infecciosa foram submetidos a raspados corneanos estudados para gram, Giemsa, cultura e qPCR (grupo de estudo). Foram avaliados fatores de risco e epidemiológicos. O grupo controle foi composto por 25 casos de úlcera dendrítica típica por herpes analisados por qPCR. Resultados: Do grupo de estudo (n=65), nove pacientes (13,8%) apresentaram cultura, qPCR e raspado negativos. Cinquenta e seis (86,2%) pacientes apresentaram cultura positiva, 51 para bacteria, 4 para fungo e 1 para ameba. A qPCR identificou 10 pacientes do grupo de cultura positiva para bactéria que também foram positivos para vírus, um VZV e 9 para HSV-1. Dos 25 pacientes que compunham o grupo controle, 21 apresentaram qPCR positivo para HSV-1. Conclusão: Herpes pode estar presente em pacientes com úlceras de córnea bacterianas e a qPCR pode ser útil na sua detecção.

Hepatitis E virus is responsible for acute and chronic liver infections worldwide. Swine hepatitis E virus has been isolated in Brazil, and a probable zoonotic transmission has been described, although data are still scarce. The aim of this study was to investigate the frequency of hepatitis E virus infection in pigs from a small-scale farm in the rural area of Parana State, South Brazil. Fecal samples were collected from 170 pigs and screened for hepatitis E virus RNA using a duplex real-time RT-PCR targeting a highly conserved 70 nt long sequence within overlapping parts of ORF2 and ORF3 as well as a 113 nt sequence of ORF2. Positive samples with high viral loads were subjected to direct sequencing and phylogenetic analysis. hepatitis E virus RNA was detected in 34 (20.0%) of the 170 pigs following positive results in at least one set of screening real-time RT-PCR primers and probes. The swine hepatitis E virus strains clustered with the genotype hepatitis E virus-3b reference sequences in the phylogenetic analysis and showed close similarity to human hepatitis E virus isolates previously reported in Brazil.

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Brazil is a non-endemic country for hepatitis E virus (HEV) infection with seroprevalence from 1% to 4% in blood donors and the general population. However, data on seroprevalence of HEV in the country are still limited. This study evaluated the prevalence of past or present HEV infection in a group of blood donors representative of the general population of the city of Sao Paulo, Southeastern Brazil. Serum samples from 500 blood donors were tested from July to September 2014 by serological and molecular methods. Anti-HEV IgG antibodies were detected in 49 (9.8%) subjects and categorized age groups revealed an age-dependent increase of HEV seroprevalence. Among the anti-HEV IgG positive subjects, only 1 had anti-HEV IgM while none tested positive for HEV-RNA. The present data demonstrate a higher seroprevalence of anti-HEV IgG than previously reported in the region. (C) 2017 Sociedade Brasileira de Infectologia. Published by Elsevier Editora Ltda. Univ Fed São Paulo, Div Infect Dis, São Paulo, SP, Brazil Fleury SA Grp, São Paulo, SP, Brazil Univ Fed São Paulo, Div Infect Dis, São Paulo, SP, Brazil FAPESP: 2012/22925-3 FAPESP: 2013/03701-0 Web of Science

Helicobacter pylori and immune thrombocytopenic purpura (ITP) association is not well established in chronic ITP (cITP) in children, although the cure of thrombocytopenia in approximately half of H. pylori eradicated adult patients has been described. The aim of this study was to investigate the effect of H. pylori eradication on platelet (PLT) recovery in cITP children and adolescents through a randomized, controlled trial. A total of 85 children (mean age 11.4 years) with cITP were prospectively enrolled. Diagnosis of H. pylori was established by two locally validated tests, (13)C-urea breath test and monoclonal stool antigen test. Twenty-two infected patients were identified, and randomly allocated into two groups: H. pylori treatment group (n = 11) and the non-intervention control group (n = 11). The control group was offered treatment if the thrombocytopenia persisted after the follow-up. At baseline, there were no differences regarding age, sex, duration of disease, and PLT count between groups. Sixty three of 85 patients were uninfected. PLT response was classified as complete response: PLT 150 × 10(9 )l(-1); partial response: PLT 50-150 × 10(9 )l(-1), or an increase of 20-30 × 10(9 )l(-1); no response: PLT 50 × 10(9 )l(-1) or an increase of20 × 10(9 )l(-1) after at least 6 months of follow-up. Complete response was observed in 60.0% (6/10, one excluded) H. pylori eradicated patients vs. 18.2% (2/11) in non-eradicated patients (p = 0.08; OR = 6.75) after 6-9 months of follow-up. Among uninfected patients, only 13.8% (8/58) presented complete response. Two non-treated controls were treated after 6-12 months of follow-up, and PLT response was observed in 61.5% (8/13) of H. pylori eradicated patients, and in 19.0% (11/58) of uninfected patients (p = 0.004). Cytotoxin associated gene A and vacuolating cytotoxin gene A IgG antibodies were present in almost all infected patients. Therefore, the study suggests that H. pylori eradication plays a role in the management of H. pylori infected cITP children and adolescents.

The chronic course of hepatitis E virus infection in immunosuppressed patients has been recently documented; however, clinical features and factors associated with this occurrence are not well known. The aim of this study was to evaluate the prevalence of previous or current HEV infection in renal transplant patients. One hundred ninety-two kidney transplant patients were studied and classified in three groups: G1-infected with hepatitis B and/or C virus; G2-patients with elevated ALT; G3-patients with normal ALT and no hepatotropic virus infection. Demographic, epidemiologic and clinical characteristics were compared between the groups. Patients with HEV infection (previous or current) were also compared to those who tested negative for HEV. HEV infection was detected using serologic (anti-HEV IgG) and molecular (HEV RNA) methods. Anti-HEV IgG was positive in 28 (15%) while HEV RNA was positive in 20 (10%). When both markers were considered, 44 (23%) patients showed evidence of previous or current HEV infection. However, both markers were concomitantly positive in only four cases (2%). In the comparative analysis, patients infected with HBV and/or HCV showed lower frequency of anti-HEV IgG (P = 0.009). There was no difference regarding demographic, epidemiologic and laboratory variable between viremic and non-viremic patients. In conclusion, past and current infection with HEV was a frequent finding among renal transplant recipients. Actively infected patients (HEV RNA positive) did not present distinct demographic and epidemiological characteristics or laboratory alterations suggestive of underlying liver damage. Therefore, infection with HEV can only be detected in immunosuppressed patients by systematic investigation of HEV RNA. J. Med. Virol. 86:948–953, 2014. © 2014 Wiley Periodicals, Inc.

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Human adenoviruses comprise an important group of etiologie agents that are responsible for various diseases in adults and children, such as respiratory, ocular, gastroenteric, and urinary infections. In immunocompromised and organ-transplanted individuals, these agents can cause generalized infections. Rapid diagnostic methods for detecting these infectious agents are not widely available.& para;& para;The aim of this work was to produce monoclonal and polyclonal anti-adenovirus antibodies to be used in a rapid diagnostic test for respiratory infections.& para;& para;Adenovirus hexons were satisfactorily purified by ultracentrifugation and chromatography. After virus purification, anti-hexon monoclonal antibodies were produced and characterized, following classical methods. Antibodies were specific for adenoviruses 2,3,5, and 41. The proposed immunochromatographic test was standardized using colloidal gold.& para;& para;The standardization of the rapid test was sufficient to detect adenovirus antigens (in nasopharyngeal lavage samples) with sensitivity of 100% and specificity of 85% when compared to direct immunofluorescence.& para;& para;The immunochromatographic assay prototype was sufficiently sensitive to detect B (3), C (2 and 5), and F (41) adenovirus samples. Although based on preliminary data, the test demonstrated the same performance as direct immunofluorescence, but with the advantage of being a point-of-care test. Further studies are still needed to confirm its effectiveness in clinical practice. (C) 2017 Sociedade Brasueira de Infectologia. Published by Elsevier Editera Ltda. Univ Fed São Paulo, Dept Med, Lab Virol, Disciplina Infectol, São Paulo, SP, Brazil Univ Fed São Paulo, Inst Biociencias, Dept Microbiol, Lab Adenovirus, São Paulo, SP, Brazil Inst Biociencias, Dept Microbiol, Lab Micol, São Paulo, SP, Brazil Inst Butantan, Lab Bacteriol, São Paulo, SP, Brazil Univ Fed São Paulo, Dept Med, Lab Virol, Disciplina Infectol, São Paulo, SP, Brazil Univ Fed São Paulo, Inst Biociencias, Dept Microbiol, Lab Adenovirus, São Paulo, SP, Brazil FAPESP: 2011/50100-6 Web of Science

Hepatitis E virus (HEV) causes acute and chronic hepatitis in organ transplant recipients. Serological evidence for HEV infection has been discovered in various population groups in Brazil, and a single acute case has been confirmed. To date, however, no cases of HEV infection in immunocompromised patients have been reported in Brazil. This study aimed to identify and characterize hepatitis E cases in renal transplant recipients in Brazil. A retrospective study was performed on 96 serum samples from renal transplant recipients with unexplained liver enzymes elevation. Three confirmed cases of HEV infection were identified that lacked seroconversion to HEV IgG antibodies. The prevalence of HEV in these patients was 3.1%. Using a sequence analysis of a 304-nucleotide fragment within ORF2, the strains were classified as genotype 3 with a low percent identity to previously characterized strains. This is the first report of hepatitis E infection in renal transplant recipients in Brazil, and the data indicate that a novel genotype 3 subvariant may be present and that further investigation is necessary to characterize the circulating HEV strains. In this setting, HEV infection should be considered as a potential cause of abnormal liver tests of unknown origin.