Risk of tuberculosis among pulmonary tuberculosis contacts: the importance of time of exposure to index cases.

INTRODUCTION: The intensity of exposure to index cases of tuberculosis [TB] may increase the risk of TB in their contacts. The aim was to determine TB risk factors among contacts of TB index cases. METHODS: A cohort study was carried out in the contacts of pulmonary TB cases registered by the epidemiological surveillance network from 01/01/2019 to 06/30/2021. The factors associated with the risk of TB in contacts were determined using the adjusted odds ratio [aOR] and its 95% confidence interval [CI]. RESULTS: From 847 TB cases, 7087 contacts were identified. The prevalence of TB was 2.0% [145/7087] and was higher in < 5 years compared to those ≥ 65 years [4.4% versus 1.2%; p < 0.001], in those exposed ≥ 6 h daily [4%], and < 6 h daily [1.6%] with respect to weekly exposure of < 6 h [0.7%; p < 0.001]. Those contacts exposed ≥ 6 h daily [aOR= 6.9; 95%CI:2.1-22.1], < 5 years [aOR= 8.3; 95%CI:1.8-37.8] and immigrants [aOR= 1.7; 95%CI:1.1-2.7] had a higher risk of TB. CONCLUSIONS: The risk of TB increases with the time of exposure to the index case and this risk is also higher in < 5 years and immigrants. Contact tracing has a high yield for detecting new cases of TB.

The impact of COVID-19 pandemic on the incidence of acute gastroenteritis outbreaks in Catalonia (Spain).

We carried out a retrospective study of acute gastroenteritis (AGE) outbreaks reported between 1 January 2015 and 31 December 2021 in Catalonia (Spain) to compare the incidence from 2015 to 2019 with that observed from 2020 to 2021. We observed a higher incidence rate of outbreaks during the prepandemic period (16.89 outbreaks/1,000,000 person-years) than during the pandemic period (6.96 outbreaks/1,000,000 person-years) (rate ratio (RR) 0.41; 95% confidence interval (CI) 0.34 to 0.51). According to the aetiology of the outbreak, those of viral aetiology decreased from 7.82 to 3.38 outbreaks/1,000,000 person-years (RR 2.31; 95% CI 1.72 to 3.12), and those of bacterial aetiology decreased from 5.01 to 2.78 outbreaks/1,000,000 person-years (RR 1.80; 95% CI 1.29 to 2.52). There was a great reduction in AGE outbreaks in Catalonia. This reduction may have been due to the effect of the nonpharmaceutical measures applied to reduce the transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), but the collapse of the healthcare system and epidemiological surveillance services may also have had a strong influence.

Factors Associated with Treatment Prescription to Pulmonary Tuberculosis Contacts in Catalonia (2019-2021): A Population-Based Epidemiological Study.

In countries with low tuberculosis (TB) incidence, the systematic testing and treatment of latent TB infection (LTBI) in contacts of pulmonary TB index cases is the standard of care. The objective of this study, conducted in Catalonia over 2019-2021, was to assess the factors associated with LTBI treatment prescription to close contacts of pulmonary TB index cases. In this population-based epidemiological study of LTBI prevalence among pulmonary TB contacts between 2019 and 2021, multiple logistic backward stepwise regression was used to identify the factors associated with treatment prescription, for which the adjusted odds ratio (aOR) and 95% confidence intervals (CI) were calculated. A total of 1487 LTBI contacts of 542 pulmonary TB index cases were studied, 80.6% of whom received a prescription. The factors associated with LTBI treatment prescription were exposure ≥6 h/day (aOR 14.20; 95% CI 5.22-38.66) and exposure <6 h/day (aOR 7.32, 95% CI 2.48-21.64), whereas the factors associated with no LTBI treatment prescription were age ≥55 years (aOR 0.22, 95% CI 0.08-0.64) and bacillus Calmette-Guerin vaccination (aOR 0.38, 95% CI 0.16-0.90). Crucial to LTBI treatment prescription is information on the contact's duration of exposure to pulmonary TB, not only for contacts exposed for ≥6 h/day, but also for contacts with lower daily exposure levels.

[Validation of the questionnaire on vaccines and hesitancy to be vaccinated in the Spanish Society of Epidemiology]. / Validación del cuestionario sobre vacunas y reticencia a vacunarse en la Sociedad Española de Epidemiología.

OBJECTIVE: To develop and validate a survey aimed at epidemiologists to measure factors associated with vaccine reluctance. METHOD: Vaccination hesitancy refers to delayed acceptance or refusal of vaccination despite the availability of vaccination services. WHO included vaccination hesitancy among the 10 global health threats in 2019. Within this conceptual framework proposed by WHO, a committee of six experts from the Spanish Society of Epidemiology (SEE) designed a self-administered questionnaire to study factors associated with vaccination hesitancy in epidemiologists. This questionnaire was approved by the SEE Board, and was sent online to all members in 2019. Based on the responses obtained, the following characteristics were validated: face validity, internal validity, construct validity, criterion validity, reliability, as well as the characteristic curves of each item and the information function per item and overall. RESULTS: The final questionnaire showed two well-defined components, perception of vaccines and confidence in the transparency of vaccine data with high degrees of fit in all aspects of validation. Both components have shown that the higher the reluctance to vaccinate the better the questionnaire reports on these aspects. CONCLUSIONS: The study has allowed the development of a validated instrument in Spanish to measure the factors associated with vaccine reluctance among epidemiologists.

Association between soluble angiotensin-converting enzyme 2 in saliva and SARS-CoV-2 infection: a cross-sectional study.

This study aimed to investigate the association between saliva soluble angiotensin-converting enzyme 2 (sACE2) and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in children and adults. We selected a convenience sample of adults with post-acute SARS-CoV-2 infection and their household children living in quarantined family households of the metropolitan Barcelona region (Spain) during the spring 2020 pandemic national lockdown. Participants were tested for saliva sACE2 quantification by western blot and nasopharyngeal SARS-CoV-2 RT-PCR detection. A total of 161 saliva samples [82 (50.9%) from children; 79 (49.1%) from females] yielded valid western blot and RT-PCR results. Saliva sACE2 was detected in 79 (96.3%) children and 76 (96.2%) convalescent adults. Twenty (24.4%) children and 20 (25.3%) convalescent adults were positive for SARS-CoV-2 in nasopharynx by RT-PCR. SARS-CoV-2 RT-PCR-negative children had a significantly higher mean proportional level of saliva sACE2 (0.540 × 10-3%) than RT-PCR-positive children (0.192 × 10-3%, p < 0.001) and convalescent adults (0.173 × 10-3%, p < 0.001). In conclusion, children negative for nasopharyngeal SARS-CoV-2 RT-PCR appear to exhibit a higher concentration of saliva sACE2 than SARS-CoV-2 RT-PCR-positive children and convalescent adults. Release of adequate levels of sACE2 in saliva could play a protective role against SARS-CoV-2.

[Towards COVID-19 control through vaccination: obstacles, challenges and opportunities. SESPAS Report 2022]. / Hacia el control de la COVID-19 a través de la vacunación: obstáculos, desafíos y oportunidades. Informe SESPAS 2022.

In Spain, the vaccination program began in a context of high transmission and low availability of vaccines. The objective of this article is to review the vaccination program against COVID-19 in Europe (3/03/2022) and assess the obstacles, challenges and opportunities posed by the control of this disease. Five vaccines are currently available in Europe: two based on mRNA technology (Comirnaty® and Spikevax®); two based on a non-replicative vector (Vaxzevria® and Janssen); and another based on subunit S (Novavax®). Health authorities have developed comprehensive vaccination strategies prioritizing the prevention of hospitalizations and deaths. In January 2022, 90% of the population was exceeded with full vaccination and 95% coverage in people over 50 years of age. The new challenge is to achieve similar coverage in the rest of the age groups. Vaccination in children and adolescents has become a priority due to the educational and social implications derived from COVID-19 in this population. Communication strategies must be renewed and access barriers eliminated to achieve good coverage. In Spain, studies have been published that find a high effectiveness of vaccination. The main strategy for controlling the pandemic and recovering social activity is the vaccination, but everything indicates that very high levels of vaccination coverage will be necessary and to follow with the non-pharmaceutical measures. In a globalized world, COVID-19 control will only be achieved with a coordinated global strategy and technical and economic support for the vaccination strategy in resource-poor countries.

Impact of COVID-19 Lockdown on the Nasopharyngeal Microbiota of Children and Adults Self-Confined at Home.

The increased incidence of COVID-19 cases and deaths in Spain in March 2020 led to the declaration by the Spanish government of a state of emergency imposing strict confinement measures on the population. The objective of this study was to characterize the nasopharyngeal microbiota of children and adults and its relation to SARS-CoV-2 infection and COVID-19 severity during the pandemic lockdown in Spain. This cross-sectional study included family households located in metropolitan Barcelona, Spain, with one adult with a previous confirmed COVID-19 episode and one or more exposed co-habiting child contacts. Nasopharyngeal swabs were used to determine SARS-CoV-2 infection status, characterize the nasopharyngeal microbiota and determine common respiratory DNA/RNA viral co-infections. A total of 173 adult cases and 470 exposed children were included. Overall, a predominance of Corynebacterium and Dolosigranulum and a limited abundance of common pathobionts including Haemophilus and Streptococcus were found both among adults and children. Children with current SARS-CoV-2 infection presented higher bacterial richness and increased Fusobacterium, Streptococcus and Prevotella abundance than non-infected children. Among adults, persistent SARS-CoV-2 RNA was associated with an increased abundance of an unclassified member of the Actinomycetales order. COVID-19 severity was associated with increased Staphylococcus and reduced Dolosigranulum abundance. The stringent COVID-19 lockdown in Spain had a significant impact on the nasopharyngeal microbiota of children, reflected in the limited abundance of common respiratory pathobionts and the predominance of Corynebacterium, regardless of SARS-CoV-2 detection. COVID-19 severity in adults was associated with decreased nasopharynx levels of healthy commensal bacteria.

Outbreaks of Gastroenteritis Due to Norovirus in Schools and Summer Camps in Catalonia, 2017-2019.

We studied outbreaks of acute gastroenteritis due to norovirus in schools and summer camps during 2017-2019 in Catalonia (Spain). The overall attack rate was 21.27% in schools and 33.42% in summer camps (RR 0.64 [95% CI 0.58-0.70]) and 52.63% of outbreaks occurred in cold months and 47.37% in warm months. The mean delay in reporting was 5.61 days (SD 5.58 days) and the mean duration was 6.11 days (SD 6.08 days), with a Pearson correlation coefficient of 0.84 (P < 0.001) between these variables. In outbreaks with person-to-person transmission, the aOR was higher the longer the delay in reporting: 3.07 (95% CI 1.21-7.81) when the delay was 5-8 days and 3.81 when it was >9 days (95% CI 1.42-10.23). The cold months posed a higher risk than the warm months. In common source outbreaks the risk was higher in children in secondary-higher education and in summer camps. IMPORTANCE Norovirus is the main cause of viral acute gastroenteritis outbreaks worldwide. The low infectious dose and the lack of long-term immunity in infected persons means that norovirus often causes outbreaks in institutions and closed and semiclosed centers. Norovirus gastroenteritis are usually mild, with no complications, but occasionally can result in hospital admission. Understanding the risk factors involved in a norovirus outbreak can reduce the spread, severity, and duration of the outbreak and, when a vaccine becomes available, this understanding would help us identify the population groups need to get vaccinated. Here, we show the outbreaks due to norovirus in schools and summer camps, the correlation between the delay in reporting and duration of outbreaks and the relationship of the attack rate and the size of the groups.

SARS-CoV-2 outbreak in a nursing home after vaccination with BNT162b2: A role for the quantification of circulating antibodies.

We describe an outbreak of SARS-CoV-2 (B.1.351) in a nursing home. At the outbreak onset 96% of residents and 76% of HCW had received two doses of BNT162b2. Twenty-eight residents (28/53) and six HCW (6/33) were infected. Infected residents had lower levels of anti-S antibodies compared to those who were not infected (157 vs 552 U/mL). Among 50 residents with available serological status, nineteen (19/25) with serum concentration < 300 U/mL and seven (7/25) with concentration > 300 U/mL acquired SARS-CoV-2 (RR 2.7 [95 %CI 1.4-5.3]). The quantification of circulating antibodies could be useful in detecting people with an impaired immune response who are at high risk of acquiring and spreading SARS-CoV-2.

Impact of the COVID-19 pandemic on contact tracing of patients with pulmonary tuberculosis.

BACKGROUND: The COVID-19 pandemic could have negative effects on tuberculosis (TB) control. The objective was to assess the impact of the pandemic in contact tracing, TB and latent tuberculosis infection (LTBI) in contacts of patients with pulmonary TB in Catalonia (Spain). METHODS: Contact tracing was carried out in cases of pulmonary TB detected during 14 months in the pre-pandemic period (1 January 2019 to 28 February 2020) and 14 months in the pandemic period (1 March 2020 to 30 April 2021). Contacts received the tuberculin skin test and/or interferon gamma release assay and it was determined whether they had TB or LTBI. Variables associated with TB or LTBI in contacts (study period and sociodemographic variables) were analyzed using adjusted odds ratio (aOR) and the 95% confidence intervals (95% CI). RESULTS: The pre-pandemic and pandemic periods showed, respectively: 503 and 255 pulmonary TB reported cases (reduction of 50.7%); and 4676 and 1687 contacts studied (reduction of 36.1%). In these periods, the proportion of TB cases among the contacts was 1.9% (84/4307) and 2.2% (30/1381) (P = 0.608); and the proportion of LTBI was 25.3% (1090/4307) and 29.2% (403/1381) (P < 0.001). The pandemic period was associated to higher LTBI proportion (aOR = 1.3; 95% CI 1.1-1.5), taking into account the effect on LTBI of the other variables studied as sex, age, household contact and migrant status. CONCLUSIONS: COVID-19 is affecting TB control due to less exhaustive TB and LTBI case detection. An increase in LTBI was observed during the pandemic period. Efforts should be made to improve detection of TB and LTBI among contacts of TB cases.

Norovirus outbreaks in long-term care facilities in Catalonia from 2017 to 2018.

Norovirus is the leading cause of outbreaks of acute viral gastroenteritis. We carried out this study to investigate outbreaks in long-term care facilities reported in 2017 and 2018 in Catalonia (Spain). The characteristics of the centers, exposed persons and the genogroups responsible were analyzed. Viral loads were estimated. The attack rate (AR) of the outbreaks studied, and the rate ratio (RR) and the odds ratio (OR) and their 95% confidence intervals as measures of association were calculated. The mean cycle thresholds were compared using the t-test for independent means. We included 30 outbreaks (4631 exposed people). The global AR was 25.93%. The RR of residents vs. staff was 2.28 (95% CI 2.0-2.6). The RR between AR in residents with total or severe dependence vs. residents with moderate, low or no-dependence was 1.23 (95% CI 1.05-1.45). The AR were higher in smaller centers than in larger ones (38.47% vs. 19.25% and RR 2; 95% CI 1.82-2.2). GII was responsible for 70% of outbreaks. No association was found between the genogroup and presenting symptoms (OR 0.96; 95% CI 0.41-2.26). Viral loads were higher in symptomatic than in asymptomatic patients (p = 0.001).

Exploring the nasopharyngeal microbiota composition in infants with whooping cough: A test-negative case-control study.

PURPOSE: The purpose of this study was to characterize the nasopharyngeal microbiota of infants with possible and confirmed pertussis compared to healthy controls. METHODS: This prospective study included all infants <1 year with microbiologically confirmed diagnosis of pertussis attended at a University Hospital over a 12-month period. For each confirmed case, up to 2 consecutive patients within the same age range and meeting the clinical case definition of pertussis but testing PCR-negative were included as possible cases. A third group of asymptomatic infants (healthy controls) were also included. Nasopharyngeal microbiota was characterized by sequencing the V3-V4 region of the 16S rRNA gene. Common respiratory DNA/RNA viral co-infection was tested by multiplex PCR. RESULTS: Twelve confirmed cases, 21 possible cases and 9 healthy controls were included. Confirmed whooping cough was primarily driven by detection of Bordetella with no other major changes on nasopharyngeal microbiota. Possible cases had limited abundance or absence of Bordetella and a distinctive microbiota with lower bacterial richness and diversity and higher rates of viral co-infection than both confirmed cases and healthy controls. Bordetella reads determined by 16S rRNA gene sequencing were found in all 12 confirmed cases (100%), 3 out of the 21 possible cases (14.3%) but in any healthy control. CONCLUSION: This study supports the usefulness of 16S rRNA gene sequencing for improved sensitivity on pertussis diagnosis compared to real-time PCR and to understand other microbial changes occurring in the nasopharynx in children <1 year old with suspected whooping cough compared to healthy controls.

The Positive Rhinovirus/Enterovirus Detection and SARS-CoV-2 Persistence beyond the Acute Infection Phase: An Intra-Household Surveillance Study.

We aimed to assess the duration of nasopharyngeal severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA persistence in adults self-confined at home after acute infection; and to identify the associations of SARS-CoV-2 persistence with respiratory virus co-detection and infection transmission. A cross-sectional intra-household study was conducted in metropolitan Barcelona (Spain) during the time period of April to June 2020. Every adult who was the first family member reported as SARS-CoV-2-positive by reverse transcription polymerase chain reaction (RT-PCR) as well as their household child contacts had nasopharyngeal swabs tested by a targeted SARS-CoV-2 RT-PCR and a multiplex viral respiratory panel after a 15 day minimum time lag. Four-hundred and four households (404 adults and 708 children) were enrolled. SARS-CoV-2 RNA was detected in 137 (33.9%) adults and 84 (11.9%) children. Rhinovirus/Enterovirus (RV/EV) was commonly found (83.3%) in co-infection with SARS-CoV-2 in adults. The mean duration of SARS-CoV-2 RNA presence in adults' nasopharynx was 52 days (range 26-83 days). The persistence of SARS-CoV-2 was significantly associated with RV/EV co-infection (adjusted odds ratio (aOR) 9.31; 95% CI 2.57-33.80) and SARS-CoV-2 detection in child contacts (aOR 2.08; 95% CI 1.24-3.51). Prolonged nasopharyngeal SARS-CoV-2 RNA persistence beyond the acute infection phase was frequent in adults quarantined at home during the first epidemic wave; which was associated with RV/EV co-infection and could enhance intra-household infection transmission.

Involvement of Workers in Closed and Semiclosed Institutions in Outbreaks of Acute Gastroenteritis Due to Norovirus.

Norovirus outbreaks frequently occur in closed or semiclosed institutions. Recent studies in Catalonia and various countries indicate that, during outbreaks in these institutions, norovirus is detected in between 23% and 60% of workers, and the prevalence of infection in asymptomatic workers involved in outbreaks ranges from 17% to 40%. In this work, we carried out a prospective study to investigate the involvement of workers in closed and semiclosed institutions during outbreaks. The attack rates (ARs) and the rate ratios (RRs) were calculated according to the type of transmission and occupational category. The RRs and 95% confidence intervals (CIs) between workers and users were calculated. The mean cycle of quantification (Cq) values were compared according to the genogroup and the presence of symptoms. ARs were higher in person-to-person transmission than in common vehicle outbreaks, and 38.8% of workers were symptomatic. The RR between workers and users was 0.46 (95% CI 0.41-0.52). The ARs in workers were high, particularly in workers with closer contact with users. The mean Cq was lower in patients than in asymptomatic infected persons, although the difference was only significant for genogroup I (GI). The frequency of asymptomatic infected persons suggests that personal hygiene measures should be followed by all workers in the centers affected.

Azithromycin to Prevent Pertussis in Household Contacts, Catalonia and Navarre, Spain, 2012-2013.

We retrospectively assessed the effectiveness of azithromycin in preventing transmission of pertussis to a patient's household contacts. We also considered the duration between symptom onset in the primary patient and azithromycin administration. We categorized contacts into 4 groups: those treated within <7 days, 8-14 days, 15-21 days, and >21 days after illness onset in the primary patient. We studied 476 primary index patients and their 1,975 household contacts, of whom 4.5% were later identified as having pertussis. When contacts started chemoprophylaxis within <21 days after the primary patient's symptom onset, the treatment was 43.9% effective. Chemoprophylaxis started >14 days after primary patient's symptom onset was less effective. We recommend that contacts of persons with pertussis begin chemoprophylaxis within <14 days after primary patient's symptom onset.

Mumps: MMR vaccination and genetic diversity of mumps virus, 2007-2011 in Catalonia, Spain.

BACKGROUND: Mumps is a vaccine-preventable disease but outbreaks have been reported in persons vaccinated with two doses of MMR vaccine. The objective was to describe the demographic features, vaccination effectiveness and genetic mumps virus diversity among laboratory-confirmed cases between 2007 and 2011 in Catalonia. METHODS: Cases and outbreaks of mumps notified to the notifiable diseases system of Catalonia between 2007 and 2011 retrospectively registered were included. Public health care centres provided written immunization records to regional public health staff to determine the vaccination history. Saliva and serum specimens were collected from suspected cases for laboratory-confirmation using real-time reverse-transcriptase PCR (rtRT-PCR) or serological testing. Phylogenetic analysis of the complete SH gene (316 nucleotides) and complete coding HN protein (1749 nucleotides) sequences was made. Categorical variables were compared using the Chi-square or Fisher's tests and continuous variables using the Student test. Vaccination effectiveness by number of MMR doses was estimated using the screening method. RESULTS: During the study period, 581 confirmed cases of mumps were notified (incidence rate 1.6 cases/100,000 persons-year), of which 60% were male. Three hundred sixty-four laboratory-confirmed cases were reported, of which 44% were confirmed by rtRT-PCR. Of the 289 laboratory-confirmed cases belonging to vaccination cohorts, 33.5% (97) had received one dose of MMR vaccine and 50% (145) two doses. Based on phylogenetic analyses of 316-nucleotide and 174-nucleotide SH sequences, the viruses belonging to viral genotypes were: genotype G (126), genotype D (23), genotype H (2), genotype F (2), genotype J (1), while one remained uncharacterized. Amino acid differences were detected between circulating strains and the Jeryl Lynn vaccine strains, although the majority of amino acid substitutions were genotype-specific. Fifty-one outbreaks were notified that included 324 confirmed mumps cases. Genotype G was the most frequent genotype detected. The family (35%), secondary schools (25%) and community outbreaks (18%) were the most frequent settings. CONCLUSIONS: Our study shows that genotype G viruses are the most prevalent in Catalonia. Most cases occurred in people who had received two doses of MMR, suggesting inadequate effectiveness of the Jeryl Lynn vaccine strain. The possible factors related are discussed.

Effectiveness and impact of the hepatitis B vaccination program in preadolescents in Catalonia 21 years after its introduction.

Hepatitis B is a viral disease of global importance. In Catalonia in the 1980s, the seroepidemiological pattern of HBV infection was low-intermediate. In 1990, the Expert Committee on Vaccinations of the Department of Health of the Generalitat of Catalonia evaluated the systematic introduction of hepatitis B vaccination in preadolescents, maintaining the vaccination of risk groups. The objective of this study was to estimate the effectiveness and impact of the systematic hepatitis B vaccination programme in preadolescents in Catalonia 21 years after its introduction. A retrospective cohort study was conducted, comparing the disease incidence in vaccinated and unvaccinated cohorts. Cases of hepatitis B were defined as those reported by the General Subdirectorate of Surveillance and Response to Public Health Emergencies between 2000 and 2014. The incidence rate was 2.5 per 100,000 persons in 1991 and 1.2 per 100,000 persons in 2014, a reduction of 52%. During the study period, 388 cases of hepatitis B infection were notified, of which three were classified as vaccine failures. Vaccine effectiveness was 99.30% (95% CI: 97.83-99.78) and the population prevented fraction in the cohorts of preadolescents studied was 64.56% (95% CI: 60.45-68.66). The effectiveness and impact of the hepatitis B vaccination program in preadolescents in Catalonia is high, with the consequent benefits for the population.

Legionella SBT applied directly to respiratory samples as a rapid molecular epidemiological tool.

Legionnaires' disease (LD) is an atypical pneumonia caused by the inhalation of Legionella. The methods used for the diagnosis of LD are direct culture of respiratory samples and urinary antigen detection. However, the sensitivity of culture is low, and the urinary antigen test is specific only for L. pneumophila sg1. Moreover, as no isolates are obtained, epidemiological studies cannot be performed. The implementation of Nested-sequence-based typing (Nested-SBT) makes it possible to carry out epidemiological studies while also confirming LD, especially in cases caused by non-sg 1. Sixty-two respiratory samples from patients with Legionella clinically confirmed by positive urinary antigen tests were cultured and tested by Nested-SBT, following the European Study Group for Legionella Infections (ESGLI) protocol. Only 2/62 (3.2%) respiratory samples were culture-positive. Amplification and sequencing of Nested-SBT genes were successfully performed in 57/62 samples (91.9%). The seven target genes were characterised in 39/57 (68.4%) respiratory samples, and the complete sequence type (ST) was obtained. The mip gene was the most frequently amplified and sequenced. Nested-SBT is a useful method for epidemiological studies in culture-negative samples, achieving a 28.7-fold improvement over the results of culture studies and reducing the time needed to obtain molecular epidemiological results.