Influenza virus type/subtype and different infection profiles by age group during 2017/2018 season

DDI-INSA em colaboração com a Rede Portuguesa de Laboratórios para o Diagnóstico da GripeBackground: Influenza has a major impact in hospitalization during each influenza season. We analysed the influenza type/subtype distribution by age group and medical care wards (ambulatory, hospital, intensive care unit). Material and Methods: During 2017/2018 season, 14 hospitals from Portugal mainland and Atlantic Island (Azores and Madeira) reported to the National Influenza Centre 13747 cases of respiratory infection, all tested for influenza type and/or subtype. Epidemiological data: age, sample collection, hospital dwelling service and patient outcome were reported. Results: From the 13747 reported cases, 3717(27%) were influenza positive of which 2033 (55%) were influenza B, 722 (19%) A unsubtyped, 505 (14%) AH3, 442 (12%) AH1pdm09 and 15(0,1%) mixed infections. Influenza A was detected in 71% (204/208) of toddlers(<5 years) although in the remaining age groups influenza B was detected in more than 50% of the confirmed flu cases. Influenza B was the predominant virus in hospitalized and ICU influenza cases between 5-14 years (69% and 75%, respectively) and played a major role in elderly (65+ years) hospitalized and ICU cases(57% and 67%, respectively). AH1pdm09 virus was detected in 30% of the influenza confirmed ICU patients, 2.1 times more than in hospitalized cases in other wards and 3.3 times more than influenza AH1pdm09 cases in ambulatory care. Influenza mixed infection were detected sporadically,mainly in hospitalized and ICU patients. From 2080 known outcomes, 40(1.9%) patients deceased, influenza was confirmed in 11(28%) of these cases. Conclusions: Cocirculation of different influenza virus type/subtype may indicate different infection profiles by age groups and should guide influenza preventive/treatment measures.N/

Influenza severe cases in hospitals, between 2014 and 2016 in Portugal

Rede Portuguesa de Laboratórios para o Diagnóstico da GripeBackground: Since 2009, the Portuguese Laboratory Network (PLNID) for Influenza Diagnosis has integrated 15 Laboratories in mainland and Atlantic Islands of Azores and Madeira. This PLNID added an important contribute to the National Influenza Surveillance Program regarding severe and hospitalized influenza cases. The present study aims to describe influenza viruses detected in influenza like illness (ILI) cases: outpatients (Outp), hospitalized (Hosp), and intensive care units (ICU), between 2014 and 2016. Methods: The PLNID performs influenza virus diagnosis by biomolecular methodologies. Weekly reports to the National Influenza Reference Laboratory ILI cases tested for influenza. Reports include data on detecting viruses, hospital assistance, antiviral therapeutics, and information on death outcome. Were reported during two winter seasons 8059 ILI cases,being 3560 cases in 2014/15 (1024 in Outp, 1750 Hosp, and 606 in ICU) and 4499 cases in 2015/2016 (1933 in Outp, 1826 Hosp, and 740 in ICU). Results: The higher percentage of influenza positive cases were detected in Outp in both seasons, 18% during 2014/15 and 20% in 2015/16. In 2014/15,influenza cases were more frequent in individuals older than 65 years old and these required more hospitalizations,even in ICU. In 2015/16,the influenza cases were mainly detected in individuals between 15-64 years old. A higher proportion of influenza positive cases with hospitalization in ICU were observed in adults between 45-64 years old.During the study period,the predominant circulating influenza viruses were different in the two seasons: influenza B and A(H3) co-circulated in 2014/15,and influenza A(H1)pdm09 was predominant during 2015/16. Even when influenza A is notthe dominant virus, A(H3) and A(H1)pdm09 subtypes correlate with higher detection rate in hospitalized cases (Hosp and UCI), with higher frequencies in adults older than 45. Influenza B,detected in higher proportion in outpatients, was frequently relatedwith influenza cases in younger age groups: 0-4 and 5-14 years old. Conclusions: This study highlights the correlation of theinfluenza virus type/subtype that circulates in each season with the possible need for hospitalization and intensive care in special groups of the population. Circulation of influenza A subtypes can cause more frequentdisease in individuals older than 45, with need of hospitalization including intensive care. On the other hand, influenza B is more frequently associated with less severe cases and with infection in children and younger adults. Influenza B circulation might predict lower number of hospitalizations.The identification of influenza type in circulation,byPLNID ineach season, could guide action planning measures in population health care.info:eu-repo/semantics/publishedVersio

Rede Portuguesa de Laboratórios para o Diagnóstico da Gripe: inverno 2013/2014

A Rede Portuguesa de Laboratórios para o Diagnóstico da Gripe (RPLDG) integra, atualmente, 15 laboratórios maioritariamente hospitalares e é coordenada pelo Laboratório Nacional de Referência para o Vírus da Gripe (LNRVG) do Departamento de Doenças Infecciosas do Instituto Nacional de Saúde Doutor Ricardo Jorge, I.P. A RPLDG realiza o diagnóstico laboratorial do vírus da gripe assim como de outros vírus respiratórios, permitindo um conhecimento mais preciso da etiologia das infeções respiratórias, particularmente em casos hospitalizados de infeção respiratória aguda grave, constituindo um complemento valioso para o PNVG. Os casos de SG provenientes de emergências hospitalares e casos de Infecção Respiratória Aguda Grave, incluindo casos com internamento em unidade de cuidados intensivos, foram notificados pelos laboratórios da Rede ao LNRVG. Dos 15 laboratórios da Rede, 13 notificaram casos de doença respiratória durante a época de 2013/2014. Os dados recolhidos foram inseridos em suporte informático tendo as bases de dados sido agregadas numa base de dados comum submetida a um processo de validação de congruência de dados. Os dados analisados correspondem ao período que decorreu entre a semana 38 de 2013 e a semana 21 de 2014. Foram notificados pelos Laboratórios da Rede um total de 3790 casos de infeção respiratória. O maior número de notificações foi observado no mês de janeiro e fevereiro (semanas 2/2014 a 8/2014), com um pico de ocorrência na semana 4/2014 com a notificação de 454 casos de infeção respiratória. O vírus da gripe foi detetado em 822 casos de infeção respiratória. O vírus influenza A foi identificado em 807 (98,2%) dos casos positivos, destes 403 (49,0%) pertencem ao subtipo A(H1)pdm09, 98 (12,0%) ao subtipo A(H3) e 306 (37,0%) vírus influenza A não foram subtipados. O vírus influenza B foi detetado em 14 (2,0%) casos. Foi identificada 1 infecção mista por vírus influenza A(H1)pdm09 e A(H3) (0,1%). A maior percentagem de casos de gripe foi observada em indivíduos entre os 15 e os 64 anos sendo o vírus influenza A(H1)pdm09 o predominantemente detetado. Nas crianças com menos de 4 anos o vírus influenza foi detetado numa proporção reduzida, apenas em 8,8% dos casos analisados laboratorialmente, sendo o agente mais detetado neste grupo etário, o vírus sincicial respiratório (dados não mostrados). A Rede Portuguesa de Laboratórios para o Diagnóstico da Gripe permitiu a deteção dos vírus da gripe em meio hospitalar, incluindo doentes em internamento e UCI. Os vírus influenza A foram predominantes e detetados em maior percentagem nos jovens e adultos

Comparación entre dos métodos de obtención de DNA a ser usados como protocolos alternativos para la detección de parásitos humanos causadores de malaria por nested PCR

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisas Básicas em Malária. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisas Básicas em Malária. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisas Básicas em Malária. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisas Básicas em Malária. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisas Básicas em Malária. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Pesquisas Básicas em Malária. Ananindeua, PA, Brasil.Um diagnóstico laboratorial correto e preciso de malária humana ainda é considerado um desafio, pois o método de referência, o da gota espessa com colocação pelo Giemsa, apresenta limitações que dificultam o controle da malária. Devido a esses problemas, várias pesquisas têm objetivado desenvolver métodos alternativos para o diagnóstico da malária. Grande parte desses estudos aborda métodos de diagnóstico molecular, que têm acarretado o desenvolvimento de algumas alternativas ao método de coloração pelo Giemsa. No entanto, esses métodos, por sua vez, apresentam suas limitações, que incluem seu alto custo, a complexidade do protocolo e a variação da qualidade das fontes de DNA e de reagentes. Neste aspecto, a técnica de nested PCR tem demonstrado ser um bom método e pode ser melhorado usando uma fonte de DNA de alta qualidade. Neste estudo foram avaliados dois métodos para a obtenção de DNA de amostras de sangue seco colhidas em papel de filtro: 1) lavagem e 2) saponina/Chelex-100. O segundo método apresentou sensibilidade e especificidade mais altas em relação ao primeiro, pois detectou mais infecções, tanto simples como mistas, bem como infecções por Plasmodium malariae. Com base nesses resultados, apresentamos o segundo como o protocolo de escolha para a obtenção de DNA. A técnica de nested PCR usando saponina/Chelex-100 para extração de DNA pode ser um método alternativo ou complementar de diagnóstico de parasitas da malária humana, mas não é considerado adequado para o uso de rotina.The correct and precise laboratory diagnosis of human malaria is still a challenge because the reference method, the Giemsa-stained thick blood smear (TS), has limitations that present problems for malaria control. Because of these problems, several studies have attempted to develop alternative methods for malaria diagnosis. Many of these studies focus on molecular diagnosis methods and have led to the development of some alternatives to TS. However, their limitations include high cost, protocol complexity and variable quality of DNA sources and reagents. Nested PCR has been shown to be a good method in this respect and it can be improved by using a high-quality source of DNA. In this study we evaluated two methods for the obtainment of DNA from dried blood samples on filter paper: 1) washing and 2) saponin/chelex-100. The second method showed higher sensitivity and specificity compared to the first, as it detected more infections, whether single or mixed, as well as Plasmodium malariae infections. Based on these results, we present this method as the protocol of choice for DNA obtainment. Nested PCR using saponin/chelex-100 for DNA extraction could be an alternative or complementary diagnosis method for human malaria parasites, but it is not appropriate for routine use

Nested PCR utilizando a gota espessa como fonte do DNA de Plasmodium: uma alternativa para esfregaços de sangue arquivados

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.The study aimed to evaluate a protocol of nested PCR using archival Giemsa-stained thick blood smears (GTS)as source of Plasmodium DNA. A total of 138 GTS from patients of five municipalities from Pará State (Amazon Region, Brazil) was included in this survey. These samples were classified in three groups (group 1: 85 Plasmodium positive and negative GTS stored in plastic box during five years; group 2: 28 Plasmodium positive and negative GTS stored in wooden box during 10 years; and group 3: 25 Trypanosoma cruzi GTS negative for Plasmodium stored in plastic box during a month) and were submitted to DNA extraction with Chelex-100. Subsequently, extracted DNA samples were quantified and the integrity was verified by electrophoresis. Nested PCR protocol was performed to detect Plasmodium species. The results of nested PCR were compared to microscopy and statistic parameters were calculated by screening test. DNA samples from all groups had acceptable quantity and purity level, but the evaluation of integrity showed 19 degraded samples from group 2. By nested PCR, this group showed very low sensitivity (29.63%) and accuracy (32.14%), while nested PCR for samples from group 1 showed 100% of sensitivity and 97.65% of accuracy. The results of this research showed that samples stored until five years can be useful as Plasmodium DNA source for nested PCR to identify Plasmodium species, being an important alternative to support retrospective studies.O estudo teve como objetivo avaliar um protocolo de nested PCR, utilizando o teste de gota espessa corada por Giemsa (GTS) como fonte de DNA de Plasmodium. Um total de 138 amostras de GTS de pacientes de cinco municípios do Estado do Pará (Região Amazônica, Brasil) foi incluído neste estudo. Essas amostras foram classificadas em três grupos (grupo 1: 85 Plasmodium GTS positivos e negativos armazenados em uma caixa de plástico durante cinco anos; grupo 2: 28 Plasmodium GTS positivos e negativos armazenados na caixa de madeira durante 10 anos; e grupo 3: 25 Trypanosoma cruzi GTS negativos para Plasmodium armazenados em caixa de plástico durante um mês) e foram submetidas à extração de DNA com Chelex-100. Subsequentemente, as amostras de DNA extraídas foram quantificadas e sua integridade foi verificada pela eletroforese. O protocolo de nested PCR foi realizado para detectar resultados das espécies de Plasmodium. Os resultados do nested PCR foram comparados com os de microscopia e os parâmetros estatísticos foram calculados pelo teste de triagem. As amostras de DNA de todos os grupos obtiveram nível, quantidade e pureza aceitáveis, mas a avaliação da integridade mostrou 19 amostras degradadas do grupo 2. Pelo nested PCR, esse grupo mostrou baixa sensibilidade (29,63%) e precisão (32,14%), enquanto que as amostras do grupo 1 apresentaram 100% de sensibilidade e 97,65% de precisão. Os resultados desta pesquisa apontam que as amostras armazenadas até cinco anos podem ser úteis como fonte de DNA de Plasmodium para que o nested PCR identifique as espécies de Plasmodium, sendo uma alternativa importante para apoiar estudos retrospectivos

Morphological atypia and molecular profile of

This study aimed to perform morphological and molecular analyses of parasites isolated from the blood of malaria-infected individuals during an outbreak in the Microregion of Cametá, State of Pará, Brazilian Amazon. A total of 260 positive samples were identified by microscopy as Plasmodium vivax; however, in three samples, forms considered unusual for the species were found and defined as morphological atypia of P. vivax. Single P. vivax infection was confirmed by qPCR in all samples. Among 256 genotyped samples, the VK247 genotype alone was identified in 255 samples, and the VK210 genotype was found in only one. The study showed that this malaria outbreak was caused by the etiological agent P. vivax, and for the first time, morphological atypia was described in isolates circulating in Brazil. Likewise, for the first time, the VK247 genotype was detected predominantly in single infections in an area of the State of Pará, which may suggest a greater circulation of the genotype in the region

Severe RSV infections in children and elderly during 2017/2018 winter season

DDI_INSA em colaboração com o DEP-INSA e a Rede Portuguesa de Laboratórios para o Diagnóstico da GripeBackground: Respiratory syncytial virus (RSV) is one of the most frequent and important respiratory viral agent that causes respiratory infection complications in younger children and elderly. RSV has an autumn / winter seasonality detected in cocirculation with influenza and other respiratory viruses. Material and Methods: During 2017/2018 season, 14 hospitals from Portugal mainland and Atlantic Island tested 4278 swabs for influenza, respiratory syncytial virus (RSV) and other respiratory viruses (oRV). Data on age and hospital service were recorded. Samples were collected from patients with mild to severe respiratory infections. Severity was correlated with the need for hospitalization. The study aimed to determine the age groups that had experienced severe RSV infections during the 2017/2018 season with the need of hospitalization, including in intensive care units (ICU). Results: Between October/2017-May/2018 were tested 4278 swabs for influenza, RSV and oRV (picornavirus, adenovirus, bocavirus, metapneumovirus, parainfluenzavirus, coronavirus). A total of 43%(1830) swabs were positive, from these 35%(639) were outpatients, 61%(1112) were hospitalized and 4% (79) were at ICU. The prevalence found were: Influenza 63%(1157), RSV 15%(266), oRV 13%(247) and 9%(160) of the cases were mixed infections. Influenza was detected in more than 70% of the positives swabs in patients aged above 15 years old. The oRV played a major role in respiratory infections in children, 0-4 and 5-14 years old, detected in 23% and 21% of the cases ,respectively. RSV was the predominant virus identified in toddlers, under 4 years old (29% of the positive samples and in 85% of codetection ). Among elderly 65+, RSV was confirmed in 13% of the respiratory infections. In hospitalized adults 65+, although influenza was detected in 80% of the positive swabs, RSV was 3.5 times more frequently detected than oRV, higher than the observed in outpatients (RSV 1.6 times more frequent than oRV). In hospitalized patients under 5 years old, RSV were detected in 31% of the positive swabs being 1.3 and 1.5 times more frequently than influenza and oRV, respectively. In ICU, 40%(32) of the cases were under 5 years old, influenza was confirmed in only 3% and RSV in 22% of the cases. 35%(28) ICU cases had 65+years old, influenza was confirmed in 57% and RSV in 14% of these patients. Conclusions: During 2017/2018, RSV was detected in severe respiratory infections. In young children (≤4 years old) RSV was the most frequently detected respiratory virus. In elderly 65+, besides influenza, RSV was frequently associated with severe respiratory infections. Prevention measures for RSV severe infections are essential not only in children but also among the elderly.N/