Clinical Perspective on Use of Long-Read Sequencing in Prenatal Diagnosis of Thalassemia.

This is an editorial focusing on the clinical perspective of a long-read sequencing method in the prenatal diagnosis of alpha- and beta-thalassemia, including a comparison between this method and standard PCR-based methods. Though incremental, the increased sensitivity and specificity using long-read sequencing is an important advantage of this methodology in the prenatal diagnostic arena due to false positive or false negative results having greater consequence when a family is making decisions about their pregnancy.

Risk of partial school closure for COVID-19 by socio-economic level in the period 2020-22.

BACKGROUND: The objective was to analyse the risk of partial school closure by economic level in Barcelona city. METHODS: In this ecological study, the risk of partial school closure for the academic years 2020-21 and 2021-22 was estimated by dividing the total number of days that each child was in quarantine or isolation by the total number of days that each child was at risk to be in quarantine or isolation in the academic year. The association between partial school closure risk and mean income by district was estimated with the Spearman rho. RESULTS: The lower the mean income, the higher the risk of partial closure (Spearman rho = 0.83; P-value = 0.003) during the academic year 2020-21. Specifically, the children from the district with the lowest income had a six times greater risk of partial school closure compared with those from the highest-income district. This risk did not show a significant socioeconomic gradient in the academic year 2021-22. CONCLUSIONS: The risk of partial school closure presented an inverse socioeconomic gradient in the city of Barcelona according to average income by district in the academic year 2020-21. This distribution was not observed in the academic year 2021-22.

A genomic meta-analysis of clinical variables and their association with intrinsic molecular subsets in systemic sclerosis.

OBJECTIVES: Four intrinsic molecular subsets (inflammatory, fibroproliferative, limited, normal-like) have previously been identified in SSc and are characterized by unique gene expression signatures and pathways. The intrinsic subsets have been linked to improvement with specific therapies. Here, we investigated associations between baseline demographics and intrinsic molecular subsets in a meta-analysis of published datasets. METHODS: Publicly available gene expression data from skin biopsies of 311 SSc patients measured by DNA microarray were classified into the intrinsic molecular subsets. RNA-sequencing data from 84 participants from the ASSET trial were used as a validation cohort. Baseline clinical demographics and intrinsic molecular subsets were tested for statistically significant associations. RESULTS: Males were more likely to be classified in the fibroproliferative subset (P = 0.0046). SSc patients who identified as African American/Black were 2.5 times more likely to be classified as fibroproliferative compared with White/Caucasian patients (P = 0.0378). ASSET participants sera positive for anti-RNA pol I and RNA pol III autoantibodies were enriched in the inflammatory subset (P = 5.8 × 10-5, P = 9.3 × 10-5, respectively), while anti-Scl-70 was enriched in the fibroproliferative subset. Mean modified Rodnan Skin Score (mRSS) was statistically higher in the inflammatory and fibroproliferative subsets compared with normal-like (P = 0.0027). The average disease duration for inflammatory subset was less than fibroproliferative and normal-like intrinsic subsets (P = 8.8 × 10-4). CONCLUSIONS: We identified multiple statistically significant differences in baseline demographics between the intrinsic subsets that may represent underlying features of disease pathogenesis (e.g. chronological stages of fibrosis) and have implications for treatments that are more likely to work in certain SSc populations.

Macrophages in Systemic Sclerosis: Novel Insights and Therapeutic Implications.

PURPOSE OF REVIEW: Macrophages play key roles in tissue homeostasis and immune surveillance, mobilizing immune activation in response to microbial invasion and promoting wound healing to repair damaged tissue. However, failure to resolve macrophage activation can lead to chronic inflammation and fibrosis, and ultimately to pathology. Activated macrophages have been implicated in the pathogenesis of systemic sclerosis (SSc), although the triggers that induce immune activation in SSc and the signaling pathways that underlie aberrant macrophage activation remain unknown. RECENT FINDINGS: Macrophages are implicated in fibrotic activation in SSc. Targeted therapeutic interventions directed against SSc macrophages may ameliorate inflammation and fibrosis. While current studies have begun to elucidate the role of macrophages in disease initiation and progression, further work is needed to address macrophage subset heterogeneity within and among SSc end-target tissues to determine the disparate functions mediated by these subsets and to identify additional targets for therapeutic intervention.

A Composite of Functional Status and Pneumonia Severity Index Improves the Prediction of Pneumonia Mortality in Older Patients.

BACKGROUND: The baseline health status may be a determinant of interest in the evolution of pneumonia. OBJECTIVE: Our objective was to assess the predictive ability of community-acquired pneumonia (CAP) mortality by combining the Barthel Index (BI) and Pneumonia Severity Index (PSI) in patients aged ≥ 65 years. DESIGN, PATIENTS AND MAIN MEASURES: In this prospective, observational, multicenter analysis of comorbidities, the clinical data, additional examinations and severity of CAP were measured by the PSI and functional status by the BI. Two multivariable models were generated: Model 1 including the PSI and BI and model 2 with PSI plus BI stratified categorically. KEY RESULTS: The total population was 1919 patients, of whom 61% had severe pneumonia (PSI IV-V) and 40.4% had some degree of dependence (BI ≤ 90 points). Mortality in the PSI V-IV group was 12.5%. Some degree of dependence was associated with increased mortality in both the mild (7.2% vs. 3.2%; p = 0.016) and severe (14% vs. 3.3%; p < 0.001) pneumonia groups. The combination of PSI IV-V and BI ≤ 90 was the greatest risk factor for mortality (aOR 4.17; 95% CI 2.48 to 7.02) in our series. CONCLUSIONS: The use of a bimodal model to assess CAP mortality (PSI + BI) provides more accurate prognostic information than the use of each index separately.

Repeated influenza vaccination for preventing severe and fatal influenza infection in older adults: a multicentre case-control study.

BACKGROUND: The effectiveness of repeated vaccination for influenza to prevent severe cases remains unclear. We evaluated the effectiveness of influenza vaccination on preventing admissions to hospital for influenza and reducing disease severity. METHODS: We conducted a case-control study in 20 hospitals in Spain during the 2013/14 and 2014/15 influenza seasons. Community-dwelling adults aged 65 years or older who were admitted to hospital for laboratory-confirmed influenza were matched with inpatient controls by sex, age, hospital and admission date. The effectiveness of vaccination in the current and 3 previous seasons in preventing influenza was estimated for inpatients with nonsevere influenza and for those with severe influenza who were admitted to intensive care units (ICUs) or who died. RESULTS: We enrolled 130 inpatients with severe and 598 with nonsevere influenza who were matched to 333 and 1493 controls, respectively. Compared with patients who were unvaccinated in the current and 3 previous seasons, adjusted effectiveness of influenza vaccination in the current and any previous season was 31% (95% confidence interval [CI] 13%-46%) in preventing admission to hospital for nonsevere influenza, 74% (95% CI 42%-88%) in preventing admissions to ICU and 70% (95% CI 34%-87%) in preventing death. Vaccination in the current season only had no significant effect on cases of severe influenza. Among inpatients with influenza, vaccination in the current and any previous season reduced the risk of severe outcomes (adjusted odds ratio 0.45, 95% CI 0.26-0.76). INTERPRETATION: Among older adults, repeated vaccination for influenza was twice as effective in preventing severe influenza compared with nonsevere influenza in patients who were admitted to hospital, which is attributable to the combination of the number of admissions to hospital for influenza that were prevented and reduced disease severity. These results reinforce recommendations for annual vaccination for influenza in older adults.

Asthma and influenza vaccination in elderly hospitalized patients: Matched case-control study in Spain.

OBJECTIVES: Influenza infection is an exacerbating factor for asthma, and its prevention is critical in managing asthmatic patients. We investigated the effect of influenza vaccination on asthmatic and non-asthmatic patients hospitalized with laboratory-confirmed influenza in Spain. METHODS: We made a matched case-control study to assess the frequency of hospitalization for influenza in people aged ≥65 years. Hospitalized patients with unplanned hospital admissions were recruited from 20 hospitals representing seven Spanish regions. Cases were defined as those hospitalized due to a laboratory-confirmed influenza infection and controls were matched by age, sex, and hospital. Data were obtained from clinical records, and patients stratified by clinical asthma history. Vaccination status and asthma due to influenza infection were analyzed according to sociodemographic variables and medical risk conditions. Multivariable analysis was made using conditional logistic regression models. RESULTS: 582 hospitalized patients with influenza (15.8% asthmatic) and 1,570 hospitalized patients without influenza (7.9% asthmatic) were included. In the multivariable conditional logistic regression using unvaccinated and non-asthmatic patients as the reference group, vaccination significantly prevented influenza in non-asthmatic patients (aOR = 0.63; 95% CI: 0.45, 0.88) and also showed a trend for a possibly protective effect in asthmatic patients (aOR = 0.79; 95% CI: 0.34, 1.81). CONCLUSION: Our results suggest that influenza vaccination could be a protective factor for asthmatic patients, although the results are inconclusive and further research is required. Practically, given the better clinical evolution of vaccinated asthma cases, and the lack of better evidence, the emphasis on vaccination of this group should continue.

Smoking may increase the risk of influenza hospitalization and reduce influenza vaccine effectiveness in the elderly.

Background: Through its effects on the immune system, smoking may facilitate influenza virus infection, its severity and its most frequent complications. The objective was to investigate the smoking history as a risk factor for influenza hospitalization and influenza vaccine effectiveness in elderly smokers/ex-smokers and non-smokers. Methods: We carried out a multicenter case-control study in the 2013-2014 and 2014-2015 influenza seasons. Cases aged ≥65 years and age-, sex-matched controls were selected from 20 Spanish hospitals. We collected epidemiological variables, comorbidities, vaccination history and the smoking history. The risk of hospitalization due to smoking (current smokers and ex-smokers) was determined using the adjusted odds ratio (aOR) with conditional logistic regression models. Vaccine effectiveness (VE) was calculated using the formula: VE = (1 - aOR) × 100. Results: We studied 728 cases and 1826 controls. Cases had a higher frequency of smoking (47.4% vs 42.1%). Smoking was associated with an increased risk of influenza hospitalization (aOR = 1.32, 95% CI: 1.04-1.68). Influenza vaccine effectiveness in preventing hospitalization was 21% (95% CI: -2 to 39) in current/ex-smokers and 39% in non-smokers (95% CI: 22-52). Conclusions: A history of smoking may increase the risk of hospitalization in smokers and ex-smokers. Preventing smoking could reduce hospitalizations due to influenza. Smokers and ex-smokers should be informed of the risk of hospitalization due to influenza infection, and encouraged to stop smoking. Smokers should be considered an at-risk group to be aggressively targeted for routine influenza vaccination.

Asthma in older people hospitalized with influenza in Spain: A case-control study.

BACKGROUND: Influenza infection is an exacerbating factor for asthma, and its prevention is critical in older patients with asthma. OBJECTIVE: This study investigated the association between asthma and influenza-related hospitalization, in Spain, of patients ages ≥ 65 years and their clinical evolution. METHODS: A multicenter case-control study was carried out in 20 Spanish hospitals during the 2013-2014 and 2014-2015 influenza seasons. Patients ages ≥ 65 years hospitalized with laboratory-confirmed influenza with and without asthma were matched with controls according to the presence of asthma, sex, age, hospital, and date of hospitalization. RESULTS: A total of 561 patients with influenza (15.9% with asthma) and 1258 patients without influenza (8.0% with asthma) were included as cases and controls, respectively. The adjusted risk of influenza for patients with asthma was calculated by multivariate conditional logistic regression. The adjustment variables were the following: smoker/nonsmoker, pneumonia in the 2 years before hospital admission, previous oral treatment with corticosteroids, influenza vaccination during the seasonal campaign, Barthel index (ordinal scale used to measure performance in activities of daily living), level of education, obesity, and the presence of other comorbidities. Patients with asthma presented a great risk of influenza (adjusted odds ratio 2.64 [95% confidence interval, 1.77-3.94]). Compared with patients without asthma, patients with asthma had more symptoms, and these had been present for longer before admission but presented a lower hospital or postdischarge mortality. CONCLUSION: This study indicated that asthma was associated with hospitalization from influenza A infection. Although patients with asthma and with influenza had more symptoms, hospital or postdischarge mortality was lower, probably due to a better response to medical treatment.

The effectiveness of influenza vaccination in preventing hospitalisations of elderly individuals in two influenza seasons: a multicentre case-control study, Spain, 2013/14 and 2014/15.

Influenza vaccination may limit the impact of influenza in the community. The aim of this study was to assess the effectiveness of influenza vaccination in preventing hospitalisation in individuals aged ≥ 65 years in Spain. A multicentre case-control study was conducted in 20 Spanish hospitals during 2013/14 and 2014/15. Patients aged ≥ 65 years who were hospitalised with laboratory-confirmed influenza were matched with controls according to sex, age and date of hospitalisation. Adjusted vaccine effectiveness (VE) was calculated by multivariate conditional logistic regression. A total of 728 cases and 1,826 matched controls were included in the study. Overall VE was 36% (95% confidence interval (CI): 22-47). VE was 51% (95% CI: 15-71) in patients without high-risk medical conditions and 30% (95% CI: 14-44) in patients with them. VE was 39% (95% CI: 20-53) in patients aged 65-79 years and 34% (95% CI: 11-51) in patients aged ≥ 80 years, and was greater against the influenza A(H1N1)pdm09 subtype than the A(H3N2) subtype. Influenza vaccination was effective in preventing hospitalisations of elderly individuals.

Factors influencing the spread of pertussis in households: a prospective study, Catalonia and Navarre, Spain, 2012 to 2013.

We aimed to investigate transmission rates of pertussis in household contacts of cases and factors associated with transmission. A prospective epidemiological study was conducted in 2012 and 2013 to determine the incidence of pertussis among household contacts of reported cases in Catalonia and Navarre, Spain. An epidemiological survey was completed for each case and contact, who were followed for 28 days to determine the source of infection (primary case) and detect the occurrence of secondary cases. Odds ratios (ORs) were used to estimate the effectiveness of vaccination and chemoprophylaxis in preventing new cases, using the formula (1 - OR) × 100. For the 688 primary cases, a total of 2,852 contacts were recorded. The household transmission rate was 16.1% (459/2,852) and rose according to the age (> 18 years) and lack of immunisation of the primary cases, and also the age (0-18 years), family relationship (siblings and children), lack of vaccination and chemoprophylaxis of contacts. Pertussis vaccine effectiveness in preventing new cases was 65.0% (95% confidence interval (CI): 11.6 to 86.2) for full vaccination (≥ 4 doses) and 59.7% (95% CI: -6.8 to 84.8) for incomplete vaccination (< 4 doses). The effectiveness of chemoprophylaxis was 62.1% (95% CI: 40.3 to 75.9). To reduce household transmission, contacts should be investigated to detect further cases and to administer chemoprophylaxis. The current vaccination status of cases and contacts can reduce household transmission.

[Primary health care professionals attitudes towards influenza immunzation in Catalonia]. / Estudio actitudes sobre vacunación antigripal en profesionales sanitarios de atención primaria de Cataluña.

OBJECTIVE: Health personnel are at risk of acquiring influenza infection and of nosocomial influenza transmission. The objective of this study was to assess the relationship between the knowledge and attitudes of primary care health personnel in Catalonia as regards influenza vaccine and the factors related to the uptake of this vaccine. DESIGN: A cross-sectional study using a web survey. SETTING: Primary care health personnel of the Catalan Health Institute. PARTICIPANTS: A total of 1212 primary health care personnel were included in the survey. Those who had medical reasons for being or not being vaccinated were excluded. RESULTS: A total of 423 replies were valid, with a 46.6% overall vaccination coverage. Vaccination rate was higher among 45 to 54 year-olds, paediatricians, those vaccinated in preceding seasons, and those living with chronic patients. There was an association between having received the vaccine and considering vaccination the best preventive action, advocating vaccination to at risk population, concern about acquiring influenza, and considering health personnel vaccination important. CONCLUSIONS: Actions taken to increase vaccination rate among health personnel should aim at correcting lack of knowledge and misconceptions about influenza vaccination of health personnel.

Influenza vaccination of primary healthcare physicians may be associated with vaccination in their patients: a vaccination coverage study.

BACKGROUND: To assess the contribution of physician-related factors, especially their influenza vaccine status, in the vaccination coverage of their patients. METHODS: A study of vaccination coverage was carried out in Spain in 2011-12. The dependent variable (vaccination coverage in patients aged ≥ 65 years) was obtained from regional records. Information was gathered on the vaccination of physicians through an anonymous web survey. We compared the vaccination coverage of patients with the vaccination of their physicians using the Student t test. Associations were determined using a multilevel regression model. RESULTS: The coverage in patients aged ≥ 65 years was 56.3% and was higher (57.3%) in patients whose physician had been vaccinated than in those whose physician had not (55.2%) (p = 0.008). In the multilevel regression model, vaccination of the physician was associated (p = 0.049) with vaccination of their patients after controlling for the effects of age (p = 0.046), region (p = 0.089), and opinions on the effectiveness of the vaccine (p = 0.013). CONCLUSIONS: Vaccination of physicians together with their opinions on the effectiveness of the vaccine may be a predictor of vaccination coverage in their patients. Further studies are required to confirm this.

Age-related differences in postural control: effects of the complexity of visual manipulation and sensorimotor contribution to postural performance.

Patterns of adaptive changes to the exposure to a sinusoidal visual stimulus can be influenced by stimulus characteristics as well as the integrity of the sensory and motor systems involved in the task. Sensorimotor deficits due to aging might alter postural responses to visual manipulation, especially in more demanding tasks. The purpose of this study was to compare postural control between young and older adults at different levels of complexity and to examine whether possible sensory and/or motor changes account for postural performance differences in older adults. Older and young adults were submitted to the following tests: postural control assessments, i.e., body sway during upright stance and induced by movement of a visual scene (moving room paradigm); sensory assessments, i.e., visual (acuity and contrast sensitivity) and somatosensory (tactile foot sensitivity and detection of passive ankle motion); and motor assessments, i.e., isometric ankle torque and muscular activity latency after stance perturbation. Older adults had worse sensory and motor performance, larger body sway amplitude during stance and stronger coupling between body sway and moving room motion than younger adults. Multiple linear regression analyses indicated that the threshold for the detection of passive ankle motion contributed the most to variances in body sway and this contribution was more striking when visual information was manipulated in a more unpredictable way. The present study suggests that less accurate information about body position is more detrimental to controlling body position, mainly for older adults in more demanding tasks.

Factors Associated with Non-Adherence to Tuberculosis Preventive Treatment among Adult Contacts of Pulmonary Tuberculosis Cases with Latent Tuberculosis Infection in Catalonia, Spain, in 2019-2021.

The aim of this study was to identify factors associated with non-adherence to tuberculosis (TB) preventive treatment among contacts with latent TB infection for new cases of pulmonary TB cases reported in Catalonia in 2019-2021. All contacts aged 18 years or older with a latent TB infection who received a TB preventive treatment were included in the study. The Chi square test and the odds ratios (OR) were used to assess the association between non-adherence to TB preventive treatment and the study variables; a p < 0.05 was considered statistically significant. Multiple logistic regression analysis was used to detect the independent factors associated with TB preventive treatment non-adherence; a p < 0.05 was considered statistically significant. The percentage of non-adherence to TB preventive treatment found in this study was 23.7%. A multivariable logistic regression analysis determined that the following factors were significantly associated with TB preventive treatment non-adherence among adult contacts: "exposure at school or workplace" (aOR = 3.34), "exposure to an index case without laboratory confirmation of TB" (aOR = 2.07), "immigrant contact" (aOR = 1.81), "male gender" (aOR = 1.75) and "exposure duration < 6 h per week or sporadic" (aOR = 1.60. By contrast, the factor "short-term TB preventive treatment regimen" (aOR = 0.38) was significantly associated with a lower treatment non-adherence. Adherence to TB preventive treatment should be improved among adult contacts of TB pulmonary cases with latent TB infection by recommending short-term treatment regimens and by developing health education activities, with a greater focus on contacts with factors associated with treatment non-adherence.

The Vaccine Training Barometer: Assessing healthcare providers' confidence to answer vaccine-related questions and their training needs.

Healthcare providers (HCP) are seen by the public as the most trustworthy source of information about vaccination. While HCPs could be a valuable partner to increase vaccine confidence in general, it is not clear whether they feel confident themselves to address questions concerning vaccination. In the context of the EU Joint Action on Vaccination (EU-JAV), the Vaccine Training Barometer, an online survey tool, was developed to assess how frequently HCPs receive questions about vaccination, how confident they feel to answer these questions, and to what extent they are willing to follow extra training. After a pilot test in Flanders, Belgium, the Barometer was launched and completed by 833 HCPs in Flanders and 291 HCPs in the Spanish regions of Catalonia, Navarre and Valencian Community from November 2020 until January 2021, during the COVID-19 pandemic, just before and during the start of the first COVID-19 vaccination campaigns. In both countries, HCPs frequently received questions about vaccination (mostly on a daily or weekly basis), and about two thirds of them indicated that the frequency of questions had increased during the three months prior to completing the survey. Most questions were about the side effects and safety of vaccines. In both countries, a considerable proportion of HCPs did not feel confident to answer vaccine-related questions (31.5% felt confident in Flanders, 21.6% in Spain). A large proportion of HCPs received questions in the last three months before the survey that they could not answer (52.4% of respondents in Flemish sample, 41.5% in Spanish sample). Only 11.4% (Flanders) and 11.3% (Spain) of the respondents felt they gained sufficient knowledge through their standard education to be able to answer questions about vaccination. Almost all respondents were willing to follow extra training on vaccination (Flanders: 95.4%, Spain: 96.6%). The Vaccine Training Barometer is thus a useful tool to monitor HCPs' confidence to answer questions about vaccination and to capture their training needs.

[Level of knowledge, attitudes and the use of preventive measures among household contacts of COVID-19 cases after the acute phase of the pandemic].

BACKGROUND: Assessment of the level of knowledge and attitudes regarding COVID-19 and preventive measures in household contacts of COVID-19 cases after the acute phase of the pandemic. METHODS: Survey among household contacts of COVID-19 cases conducted in healthcare centers in Navarre (1) and Catalonia (8) between May 2022 and July 2023. The frequency of use of preventive measures and the knowledge and attitudes towards COVID-19 were assessed through 12 items. RESULTS: 215 contacts participated who correctly used the preventive measures (>85%), except for facemasks (35.8%) and interpersonal distance (47%); >85% showed adequate knowledge (5/6 items) and >80% had a positive attitude (3/6 items). Moreover, 54.7% considered that COVID-19 negatively affected their life and 54.1% that it is better to develop immunity by getting infected than by vaccination. CONCLUSIONS: Household contacts show a correct level of knowledge and positive attitude towards the disease and its preventive measures.

Absence of short-term changes in knowledge and attitudes among household contacts of COVID-19 cases during the post-acute phase of the pandemic in Catalonia and Navarre, Spain.

Objectives: To evaluate short-term changes in knowledge and attitude towards COVID-19 and preventive measures during the post-acute phase of the pandemic in Spain. Methods: A survey was performed in Catalonia and Navarre between May-2022 and July-2023 in household contacts of COVID-19 cases. Knowledge and attitude were assessed at baseline and at three months, using a Likert scale. Responses were grouped according to correct or incorrect. Results: At baseline, 172 subjects were contacted, 118 (69%) of which completed a follow-up interview three months later. Knowledge of correct hand-washing and mask protocols had maintained over time (-1.7%, p = 0.553 and - 2.5%, p = 0.473, respectively). Attitudes toward preventive measures was adequate in the first interview (86%), but attitudes regarding use of face masks decreased significantly (-9.1%, p = 0.048) over time in participants with higher risk of severe COVID-19. However, most short-term changes in knowledge and attitudes were not statistically significant. Conclusion: Household contacts showed correct knowledge and attitude towards COVID-19 and its preventive measures, without significant changes in the short term despite a relaxation of government-mandated preventive measures. These results provide relevant information in case of a new health emergency due to respiratory viruses.

Assessment of Long-Term Changes in Knowledge and Attitudes of Household Contacts of COVID-19 Cases in Northern Spain.

This study aims to describe the long-term changes in the knowledge of, and attitudes towards, COVID-19 and its preventive measures in northern Spain. A telephonic survey was performed among household contacts of COVID-19 cases in Catalonia and Navarre between May 2022 and December 2023. Knowledge and attitudes were assessed through 12 questions using a Likert scale, and responses were grouped as correct or incorrect. The change from baseline to the 6-month follow-up was evaluated with the absolute difference (AD) using the proportion of correct answers. At baseline, 299 subjects were contacted, of whom 63.2% (189) completed the 6-month follow-up. Correct knowledge of transmission (>85%) and the use of preventive measures (>92%) were observed at baseline and maintained over time. The attitudes towards face mask use remained adequate over the course of six months (>79%). However, attitudes regarding the use of face masks indoors (AD = -16.4%; p < 0.001) and those who thought that COVID-19 had a negative impact on their lives (AD = -16.5%; p < 0.001) decreased after 6 months. In the post-acute phase of the pandemic, household contacts maintained the correct level of knowledge towards COVID-19, while some attitudes decreased. These results should serve as a guide for health policy makers in decision-making in case of a new increase in the incidence of SARS-CoV-2.

Analytical validation of a semi-automated methodology for quantitative measurement of SARS-CoV-2 RNA in wastewater collected in northern New England.

Wastewater-based epidemiology (WBE) can be used to monitor the community presence of infectious disease pathogens of public health concern such as the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Viral nucleic acid has been detected in the stool of SARS-CoV-2-infected individuals. Asymptomatic SARS-CoV-2 infections make community monitoring difficult without extensive and continuous population screening. In this study, we validated a procedure that includes manual pre-processing, automated SARS-CoV-2 RNA extraction and detection workflows using both reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR) and reverse transcriptase droplet digital PCR (RT-ddPCR). Genomic RNA and calibration materials were used to create known concentrations of viral material to determine the linearity, accuracy, and precision of the wastewater extraction and SARS-CoV-2 RNA detection. Both RT-qPCR and RT-ddPCR perform similarly in all the validation experiments, with a limit of detection of 50 copies/mL. A wastewater sample from a care facility with a known outbreak was assessed for viral content in replicate, and we showed consistent results across both assays. Finally, in a 2-week survey of two New Hampshire cities, we assessed the suitability of our methods for daily surveillance. This paper describes the technical validation of a molecular assay that can be used for long-term monitoring of SARS-CoV-2 in wastewater as a potential tool for community surveillance to assist with public health efforts.IMPORTANCEThis paper describes the technical validation of a molecular assay that can be used for the long-term monitoring of SARS-CoV-2 in wastewater as a potential tool for community surveillance to assist with public health efforts.