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- Cross-protection to new drifted influenza A(H3) viruses and prevalence of protective antibodies to seasonal influenza, during 2014 in PortugalPublication . Guiomar, Raquel; Pereira da Silva, Susana; Conde, Patrícia; Cristóvão, Paula; Maia, Ana Carina; Pechirra, Pedro; Rodrigues, Ana Paula; Nunes, Baltazar; Milho, Luís; Coelho, Ana Paula; Fernandes, Aida; Caseiro, Paula; Rodrigues, Fernando; Correia, Lurdes; Pereira-Vaz, João; Almeida, Sofia; Branquinho, Paula; Côrte-Real, Rita; Viseu, Regina; Peres, Maria João; Sanches, Raquel; Dantas, Filipa; Freitas, Ludovina; Andrade, Graça; Maurílio, Manuel; Caldeira, Filomena; Cabral Veloso, Rita; Mota-Vieira, Luísa; Soares, Marta; Couto, Ana Rita; Bruges-Armas, Jácome; Mouro Pinto, Rita; Sobrinho Simões, Joana; Rosário Costa, Maria; Guimarães, João Tiago; Martins, Luís; Cunha, MárioIntroduction: Immune profile for influenza viruses is highly changeable over time. Serological studies can assess the prevalence of influenza, estimate the risk of infection, highlight asymptomatic infection rate and can also provide data on vaccine coverage. The aims of the study were to evaluate pre-existing cross-protection against influenza A(H3) drift viruses and to assess influenza immunity in the Portuguese population. Materials and methods: We developed a cross-sectional study based on a convenience sample of 626 sera collected during June 2014, covering all age groups, both gender and all administrative health regions of Portugal. Sera antibody titers for seasonal and new A(H3) drift influenza virus were evaluated by hemagglutination inhibition assay (HI). Seroprevalence to each seasonal influenza vaccine strain virus and to the new A(H3) drift circulating strain was estimated by age group, gender and region and compared with seasonal influenza-like illness (ILI) incidence rates before and after the study period. Results: Our findings suggest that seroprevalences of influenza A(H3) (39.9%; 95% CI: 36.2–43.8) and A(H1)pdm09 (29.7%; 95% CI: 26.3–33.4) antibodies were higher than for influenza B, in line with high ILI incidence rates for A(H3) followed by A(H1)pdm09, during 2013/2014 season. Low pre-existing crossprotection against new A(H3) drift viruses were observed in A(H3) seropositive individuals (46%). Both against influenza A(H1)pdm09 and A(H3) seroprotection was highest in younger than 14-years old. Protective antibodies against influenza B were highest in those older than 65 years old, especially for B/Yamagata lineage, 33.3% (95% CI: 25.7–41.9). Women showed a high seroprevalence to influenza, although without statistical significance, when compared to men. A significant decreasing trend in seroprotection from north to south regions of Portugal mainland was observed. Conclusions: Our results emphasize that low seroprotection increases the risk of influenza infection in the following winter season. Seroepidemiological studies can inform policy makers on the need for vaccination and additional preventive measures.
- Cross-protection to new drifted influenza A(H3) viruses and prevalence of protective antibodies to seasonal influenza, during 2014 in PortugalPublication . Guiomar, Raquel; Pereira da Silva, Susana; Conde, Patrícia; Cristóvão, Paula; Maia, Ana Carina; Pechirra, Pedro; Rodrigues, Ana Paula; Nunes, Baltazar; Milho, Luís; Coelho, Ana Paula; Fernandes, Aida; Caseiro, Paula; Rodrigues, Fernando; Correia, Lurdes; Pereira-Vaz, João; Almeida, Sofia; Branquinho, Paula; Côrte-Real, Rita; Viseu, Regina; Peres, Maria João; Sanches, Raquel; Dantas, Filipa; Freitas, Ludovina; Andrade, Graça; Maurílio, Manuel; Caldeira, Filomena; Cabral Veloso, Rita; Mota-Vieira, Luísa; Soares, Marta; Couto, Ana Rita; Bruges-Armas, Jácome; Mouro Pinto, Rita; Sobrinho Simões, Joana; Costa, Maria do Rosário; Guimarães, João Tiago; Martins, Luís; Cunha, MárioIntroduction: Immune profile for influenza viruses is highly changeable over time. Serological studies can assess the prevalence of influenza, estimate the risk of infection, highlight asymptomatic infection rate and can also provide data on vaccine coverage. The aims of the study were to evaluate pre-existing cross-protection against influenza A(H3) drift viruses and to assess influenza immunity in the Portuguese population. Materials and methods: We developed a cross-sectional study based on a convenience sample of 626 sera collected during June 2014, covering all age groups, both gender and all administrative health regions of Portugal. Sera antibody titers for seasonal and new A(H3) drift influenza virus were evaluated by hemagglutination inhibition assay (HI). Seroprevalence to each seasonal influenza vaccine strain virus and to the new A(H3) drift circulating strain was estimated by age group, gender and region and compared with seasonal influenza-like illness (ILI) incidence rates before and after the study period. Results: Our findings suggest that seroprevalences of influenza A(H3) (39.9%; 95% CI: 36.2-43.8) and A(H1)pdm09 (29.7%; 95% CI: 26.3-33.4) antibodies were higher than for influenza B, in line with high ILI incidence rates for A(H3) followed by A(H1)pdm09, during 2013/2014 season. Low pre-existing cross-protection against new A(H3) drift viruses were observed in A(H3) seropositive individuals (46%). Both against influenza A(H1)pdm09 and A(H3) seroprotection was highest in younger than 14-years old. Protective antibodies against influenza B were highest in those older than 65 years old, especially for B/ Yamagata lineage, 33.3% (95% CI: 25.7-41.9). Women showed a high seroprevalence to influenza, although without statistical significance, when compared to men. A significant decreasing trend in seroprotection from north to south regions of Portugal mainland was observed. Conclusions: Our results emphasize that low seroprotection increases the risk of influenza infection in the following winter season. Seroepidemiological studies can inform policy makers on the need for vaccination and additional preventive measures.
- Epidemiology and genetic variability of respiratory syncytial virus in Portugal, 2014-2018Publication . Sáez-López, Emma; Cristóvão, Paula; Costa, Inês; Pechirra, Pedro; Conde, Patrícia; Guiomar, Raquel; Peres, Maria João; Viseu, Regina; Lopes, Paulo; Soares, Vânia; Vale, Fátima; Fonseca, Patricia; Freitas, Ludivina; Alves, Jose; Pessanha, Maria Ana; Toscano, Cristina; Mota-Vieira, Luísa; Veloso, Rita Cabral; Côrte-Real, Rita; Branquinho, Paula; Pereira‑Vaz, João; Rodrigues, Fernando; Cunha, Mário; Martins, Luís; Mota, Paula; Couto, Ana Rita; Bruges-Armas, Jácome; Almeida, Sofia; Rodrigues, Débora; Portuguese Laboratory Network for the Diagnosis of Influenza InfectionIntroduction: Respiratory syncytial virus (RSV) is associated with substantial morbidity and mortality since it is a predominant viral agent causing respiratory tract infections in infants, young children and the elderly. Considering the availability of the RSV vaccines in the coming years, molecular understanding in RSV is necessary. Objective: The objective of the present study was to describe RSV epidemiology and genotype variability in Portugal during the 2014/15-2017/18 period. Material and methods: Epidemiological data and RSV-positive samples from patients with a respiratory infection were collected through the non-sentinel and sentinel influenza surveillance system (ISS). RSV detection, subtyping in A and B, and sequencing of the second hypervariable region (HVR2) of G gene were performed by molecular methods. Phylogenetic trees were generated using the Neighbor-Joining method and p-distance model on MEGA 7.0. Results: RSV prevalence varied between the sentinel (2.5%, 97/3891) and the non-sentinel ISS (20.7%, 3138/16779), being higher (P < 0.0001) among children aged <5 years. Bronchiolitis (62.9%, 183/291) and influenza-like illness (24.6%, 14/57) were associated (P < 0.0001) with RSV laboratory confirmation among children aged <6 months and adults ≥65 years, respectively. The HVR2 was sequenced for 562 samples. RSV-A (46.4%, 261/562) and RSV-B (53.6%, 301/562) strains clustered mainly to ON1 (89.2%, 233/261) and BA9 (92%, 277/301) genotypes, respectively, although NA1 and BA10 were also present until 2015/2016. Conclusion: The sequence and phylogenetic analysis reflected the relatively high diversity of Portuguese RSV strains. BA9 and ON1 genotypes, which have been circulating in Portugal since 2010/2011 and 2011/2012 respectively, predominated during the whole study period.
- Influenza seroprotection correlates with predominant circulating viruses during 2014/15 and 2015/16 seasons in PortugalPublication . Guiomar, Raquel; Cristóvão, Paula; Conde, Patrícia; Costa, Inês; Pechirra, Pedro; Rodrigues, Ana Paula; Pereira da Silva, Susana; Nunes, Baltazar; Mouro Pinto, Rita; Sobrinho Simões, Joana; Costa, Maria do Rosário; Guimarães, João Tiago; Rodrigues, Fernando; Correia, Lurdes; Pereira-Vaz, João; Caseiro, Paula; Cabral Veloso, Rita; Mota Vieira, Luísa; Pimentel Couto, Ana Rita; Santos, Margarida; Bruges Armas, Jácome; Branquinho, Paula; Corte-Real, Rita; Martins, Luís; Cunha, Mário; Almeida, Sofia; Viseu, Regina; Inácio, Filipe; Peres, Maria João; Milho, Luís; Fernandes, Aida; Maurílio, Manuel; Caldeira, Filomena; Sanches, Raquel; Dantas, Filipa; Freitas, Ludivina; Andrade, Graça; Mota, PaulaBACKGROUND: Population immune profile for influenza is highly affected by circulating influenza viruses, thus changing the risk of infection for influenza. This study aims to assess influenza immunity in the Portuguese population by age groups, during 2014 and 2015 and establish a relationship between seroprotection and circulating influenza viruses in 2014/15 and 2015/16 seasons. METHODS: Two cross-sectional studies were developed based on a convenience serum sample collected in June 2014 (n=626) and July 2015 (n=675) in hospitals from mainland and Azores and Madeira.Serums equally represent all age groups. Antibody titers were evaluated by HI assay for strains recommended for seasonal influenza vaccine northern hemisphere,2014/15 and 2015/2016. Seroprevalences were estimated for each strain by age group and the association with seasonal cumulative influenza-like illness (ILI) rates for influenza virus during both seasons was analised. RESULTS: In June 2014 the highest seroprotection was observed for influenza A(H3) (39.0%; 95% CI: 36.2-43.8%) and A(H1)pdm09 (29.7; 95% CI: 26.3-33.4%), with higher levels in children 5-14 years old. In 2014/2015 a dominant circulation of influenza B/Yamagata was observed with high incidence rates in individuals under 65 years old, the ones that had lower seroprotection. Although before the start of the season high protection for A(H3) was observed, the circulation of the new drift A(H3) strains had gained an immunological advantage,in accordance with A(H3) elevated incidence rates observed during 2014/15. In July 2015 the highest seroprotection was observed for influenza B/ Yamagata (55.1%; 95% CI: 51.4-58.9%), 2.4 times the estimated 2014.This increase was even more pronounced in younger (≤ 4 years old), 6.3 times increase in 2015.This fact is in agreement with the predominant influenza B virus detected and the high ILI incidence rate observed in children during 2014/2015 epidemic. Seroprotection levels for influenza A in July 2015 were not significantly different from 2014.During 2015/16 season, influenza A(H1N1)pdm09 was predominant, with high incidence rate in < 65 year old. Influenza B/Victoria lineage,although detected at low levels increased in frequency, in agreement with the lowest level of seroprotection detected in the general population before the start of 2015/2016 season (21.8%; 95% CI: 18.7-24.0%). CONCLUSIONS There was a correlation between virus circulation, incidence rates for each age group and the previous seroprotection for seasonal influenza viruses.Our study highlights the value of measuring the serological profile for influenza to establishe risk groups for infection for which an increase preventive measures, including vaccination, should be fostered.
- Influenza severe cases in hospitals, between 2014 and 2016 in PortugalPublication . Guiomar, Raquel; Pechirra, Pedro; Cristóvão, Paula; Costa, Inês; Conde, Patrícia; Corte-Real, Rita; Branquinho, Paula; Silvestre, Maria José; Almeida Santos, Madalena; Fernandes, Isabel; Dias, Isabel; Rodrigues, Sónia; Sena, Nadir; Lazzara, Daniela; Sobrinho Simões, Joana; Costa, Maria do Rosário; Guimarães, João Tiago; Rodrigues, Fernando; Pereira-Vaz, João; Correia, Lurdes; Andrade, Graça; Freitas, Ludivina; Figueira, Neuza; Sanches, Raquel; Marques, Mónica; Barros, Margarida; Mota Vieira, Luísa; Cabral Veloso, Rita; Castelo Branco, Cláudia; Pimentel, Sílvia; Duarte, Joana; Pereirinha, Tânia; Bulhões, Sara; Moniz, Raquel; Brilhante, Maria José; Bruges Armas, Jácome; Pimentel Couto, Ana Rita; Santos, Margarida; Soares, Marta; Melo Cristino, José; Ribeiro, Carlos; Carvalho, Dinah; Barreto, Rosário; Ramos, Maria Helena; Castro, Ana Paula; Matos Santos, Ana Cláudia; Cunha, Mário; Martins, Luís; Almeida, Sofia; Peres, Maria João; Viseu, Regina; Inácio, Filipe; Mota, PaulaBackground: 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.
- Influenza virus type/subtype and different infection profiles by age group during 2017/2018 seasonPublication . Guiomar, Raquel; Pechirra, Pedro; Cristóvão, Paula; Costa, Inês; Conde, Patrícia; Côrte-Real, Rita; Branquinho, Paula; Garcia, David; Conde, Sílvia; Rodrigues, Fernando; Pereira-Vaz, João; Alves, José; Freitas, Ludivina; Mota Vieira, Luísa; Cabral Veloso, Rita; Bruges Armas, Jácome; Couto, Ana Rita; Ribeiro, Carlos; Barreto, Rosário; Cunha, Mário; Martins, Luís; Almeida, Sofia; Peres, Maria João; Viseu, Regina; Mota, Paula; Lopes, Paulo; Soares, Vânia; Vale, Fátima; Fonseca, Patrícia; Toscano, Cristina; Dias, AnaBackground: 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.
- Molecular characterization of respiratory syncycial virus during 2015-2016 season in PortugalPublication . Cristóvão, Paula; Pereira, Diogo; Pechirra, Pedro; Pereira-Vaz, João; Correia, Lurdes; Rodrigues, Fernando; Andrade, Graça; Corte-Real, Rita; Branquinho, Paula; Peres, Maria João; Viseu, Regina; Balseiro, Maria Jesus; Mota, Paula; Guiomar, RaquelBackground: 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. Genetic diversity in both of RSV A and B subtypes increased in last years with the spread of new genotypes. This study aims to describe the genetic variability of RSV during 2015/2016 season in Portugal and correlate the circulating genotypes with detected ones in previous seasons. Will also be evaluated the association between genotype, clinical diagnosis and age. Methods: During 2015/16 winter season, between November/2015 and February/2016, 45 RSV were genetically characterized. RSV positive respiratory samples were collected in two settings: children under 5 years old diagnosed by hospital laboratories from the Portuguese Laboratory Network for the Diagnosis of Influenza Infection, and all age Influenza-like illness (ILI) patients reported by primary care health services and diagnosed by the National Influenza Reference Laboratory. All samples were irreversibly anonymized. Demographic and clinical data were collected. RSV detection was performed by real-time PCR and other biomolecular methods. RSV genotype was assigned by the nucleotide sequence of the hypervariable C-terminal region of the G protein gene and the phylogenetic analysis was performed in MEGA 6.0. Results: From 45 RSV genetically characterized, 31 (69%) were reported by hospitals, patients age ranged from newborn to 4 years old. From these, 25 (81%; 25/31) patients were hospitalized, being the bronchiolitis the most frequent diagnosis. While 14 (31%) RSV cases came from primary care health services, patients age ranged from 3 to 83 and all had a clinical diagnosis of ILI. Were included patients from both genders in equal proportions. RSV A and B co-circulated during 2015/2016 season. Were genetically characterized 21 (47%) RSV A and 24 (53%) RSV B. 90% (19/21) of RSV A clustered in ON1 genotype, the others 2 clustered with NA1 genotype. All RSV B present a BA-like genotype, 70% (17/24) were similar to BA9 and 30% (7/24) clustered with BA10 genotype. Conclusions: During 2015/2016 season was observed a co-circulation of RSVA and RSVB. In present study ON1genotype was predominant in circulation among RSVA, this was also detected as the major RSVA genotype at the global level. Only two RSVA belonged to NA1 genotype. In Portugal, NA1 was in circulation during 2010-2012 period. Undetected since 2012, it seems to reappear during 2015/16 season. All RSVB characterized belonged to BA genotypes, the majority clustered within BA9 genotype. BA10 genotype was also identified in circulation at low frequency. BA9 and BA10 were being found in co-circulation since 2011/12. No association was found between age, clinical diagnosis and RSVA and B genotypes. RSV has an important impact in children in high-risk groups highlighting the need off a continuous RSV surveillance each winter.
- Severe RSV infections in children and elderly during 2017/2018 winter seasonPublication . Guiomar, Raquel; Pechirra, Pedro; Costa, Inês; Cristóvão, Paula; Conde, Patrícia; Nunes, Baltazar; Rodrigues, Ana Paula; Côrte-Real, Rita; Branquinho, Paula; Garcia, David; Conde, Sílvia; Rodrigues, Fernando; Pereira-Vaz, João; Alves, José; Ludivina, Freitas; Mota Vieira, Luísa; Cabral Veloso, Rita; Bruges Armas, Jácome; Couto, Ana Rita; Ribeiro, Carlos; Barreto, Rosário; Cunha, Mário; Martins, Luís; Almeida, Sofia; Peres, Maria João; Viseu, Regina; Mota, Paula; Lopes, Paulo; Soares, Vânia; Vale, Fátima; Fonseca, Patrícia; Toscano, Cristina; Dias, AnaBackground: 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.