Bacterial Cross-Transmission between Inanimate Surfaces and Patients in Intensive Care Units under Real-World Conditions: A Repeated Cross-Sectional Study.

Background/Objectives: Contaminated surfaces play an important role in the nosocomial infection of patients in intensive care units (ICUs). This study, conducted in two ICUs at Edouard Herriot Hospital (Lyon, France), aimed to describe rooms' microbial ecology and explore the potential link between environmental contamination and patients' colonization and/or infection. Methods: Environmental samples were realized once monthly from January 2020 to December 2021 on surfaces close to the patient (bedrails, bedside table, and dedicated stethoscope) and healthcare workers' high-touch surfaces, which were distant from the patient (computer, worktop/nurse cart, washbasin, and hydro-alcoholic solution/soap dispenser). Environmental bacteria were compared to the cultures of the patients hospitalized in the sampled room over a period of ± 10 days from the environmental sampling. Results: Overall, 137 samples were collected: 90.7% of the samples close to patients, and 87.9% of the distant ones were positives. Overall, 223 bacteria were isolated, mainly: Enterococcus faecalis (15.7%), Pantoea agglomerans (8.1%), Enterobacter cloacae/asburiae (6.3%), Bacillus cereus and other Bacillus spp (6.3%), Enterococcusfaecium (5.8%), Stenotrophomonas maltophilia (5.4%), and Acinetobacter baumannii (4.9%). Throughout the study, 142 patients were included, of which, n = 67 (47.2%) were infected or colonized by at least one bacterium. In fourteen cases, the same bacterial species were found both in environment and patient samples, with the suspicion of a cross-contamination between the patient-environment (n = 10) and environment-patient (n = 4). Conclusions: In this work, we found a high level of bacterial contamination on ICU rooms' surfaces and described several cases of potential cross-contamination between environment and patients in real-world conditions.

The MTB/MDR ELITe MGB® Kit: Performance Assessment for Pulmonary, Extra-Pulmonary, and Resistant Tuberculosis Diagnosis, and Integration in the Laboratory Workflow of a French Center.

A rapid and reliable diagnostic for tuberculosis, including the detection of both rifampicin (RIF) and isoniazid (INH) resistance, is essential for appropriate patient care. Nucleic acid amplification tests are a fast alternative to methods based on Mycobacterium tuberculosis complex (MTB) cultures. Thus, the performance of the MDR/MTB ELITe MGB® Kit on the ELITe InGenius® platform was retrospectively evaluated for MTB detection on pulmonary and extra-pulmonary samples and for RIF/INH resistance detection on MTB strains. The sensitivity and specificity of the kit for MTB detection compared to the MTB culture were 80.0% and 100.0%, respectively. For the antimicrobial susceptibility prediction, the agreement with phenotypic antimicrobial susceptibility testing (AST) was 92.0%. For RIF, the sensitivity was 100.0% and the specificity was 95.5%. For INH, the sensitivity and specificity were 75.0% and 100.0%, respectively. A single RIF false-positive result was obtained for a strain with a low level of RIF resistance that was not detected by phenotypic AST, but carrying a rpoB L452P mutation. INH false-negative results (3) were due to mutations on the katG gene that were not probed by the test. Overall, the MTB/MDR ELITe MGB® Kit presents a strong performance for MTB detection and for the detection of both RIF and INH resistance, with an easy integration in laboratory workflow thanks to its fully automatized system.

Prospective Whole-Genome Sequencing in Tuberculosis Outbreak Investigation, France, 2017-2018.

During June 2017-April 2018, active tuberculosis with Beijing SIT1 isolates was diagnosed in 14 persons living in 4 distant cities in France. Whole-genome sequencing indicated that these patients belonged to a single transmission chain. Whole-genome sequencing-based laboratory investigations enabled prompt tracing of linked cases to improve tuberculosis control.

Evaluation of Xpert MTB/RIF Ultra performance for pulmonary tuberculosis diagnosis on smear-negative respiratory samples in a French centre.

Tuberculosis (TB) is a worldwide public health concern, including in high-resource countries with a low prevalence of TB. Xpert MTB/RIF assay was developed to improve TB and rifampicin (RIF) resistance detection, but sensitivity remains poor on smear-negative sputum. Xpert MTB/RIF Ultra assay was designed to enhance the sensitivity of TB detection in clinical samples. Herein, we evaluated retrospectively the performance of this test on smear-negative respiratory samples. Respiratory specimens with smear-negative and a Mycobacterium tuberculosis (MTB) complex-positive culture were retrospectively selected from those taken from patients during routine care, and analysed in the Mycobacteria Laboratory of the Lyon University hospital, France. Specimens were stored at - 20 °C before testing by Xpert MTB/RIF Ultra. For each sample, growth delay and date of anti-TB treatment initiation were recorded. Forty-six samples-29 sputum, 8 bronchial aspirates, 6 broncho-alveolar lavages, and 3 gastric aspirates-were selected. Among samples collected before treatment initiation (n = 33), sensitivity was 81.8% (95% CI [64.5; 93.0]) and there was a significant correlation between the quantitative measurements (Ct) of Xpert MTB/RIF Ultra assay and the time to growth detection in culture. Among samples collected after treatment initiation (n = 12), sensitivity was 100%, without correlation with time to growth detection due to presence of afterglow DNA in samples. In high-resource settings, the Xpert MTB/RIF Ultra test represents a useful tool for pulmonary TB diagnosis, notably for the paucibacillary forms. Moreover, quantitative measurement of Xpert MTB/RIF Ultra could help to predict time to MTB culture positivity and be used as a quality indicator of MTB culture process.

Changing patterns of human migrations shaped the global population structure of Mycobacterium tuberculosis in France.

Mycobacterium tuberculosis (Mtb) exhibits a structured phylogeographic distribution worldwide linked with human migrations. We sought to infer how the interactions between distinct human populations shape the global population structure of Mtb on a regional scale. We applied the recently described timescaled haplotypic density (THD) technique on 638 minisatellite-based Mtb genotypes from French tuberculosis patients. THD with a long-term (200 y) timescale indicated that Mtb population in France had been mostly influenced by interactions with Eastern and Southern Europe and, to a lesser extent, Northern and Middle Africa, consistent with historical migrations favored by geographic proximity or commercial exchanges with former French colonies. Restricting the timescale to 20 y, THD identified a sustained influence of Northern Africa, but not Europe where tuberculosis incidence decreased sharply. Evolving interactions between human populations, thus, measurably influence the local population structure of Mtb. Relevant information on such interactions can be inferred using THD from Mtb genotypes.

Strain-specific estimation of epidemic success provides insights into the transmission dynamics of tuberculosis.

The transmission dynamics of tuberculosis involves complex interactions of socio-economic and, possibly, microbiological factors. We describe an analytical framework to infer factors of epidemic success based on the joint analysis of epidemiological, clinical and pathogen genetic data. We derive isolate-specific, genetic distance-based estimates of epidemic success, and we represent success-related time-dependent concepts, namely epidemicity and endemicity, by restricting analysis to specific time scales. The method is applied to analyze a surveillance-based cohort of 1,641 tuberculosis patients with minisatellite-based isolate genotypes. Known predictors of isolate endemicity (older age, native status) and epidemicity (younger age, sputum smear positivity) were identified with high confidence (P < 0.001). Long-term epidemic success also correlated with the ability of Euro-American and Beijing MTBC lineages to cause active pulmonary infection, independent of patient age and country of origin. Our results demonstrate how important insights into the transmission dynamics of tuberculosis can be gained from active surveillance data.

Factors associated with Vancomycin-resistant Enterococcus acquisition during a large outbreak.

Between 2013 and 2014 a Vancomycin-resistant Enterococci (VRE) outbreak occurred in a teaching hospital in France. The outbreak was significant possibly due to the lack of implementation of recommended control measures. The aim of this study was to identify the effect of the lack of adherence to control measures for prevention of VRE acquisition in contact patients taking into account individual risk factors. Contact patients (first two months of the outbreak) with VRE acquisition were compared to patients without VRE acquisition (univariate and logistic regression), in terms of institutional characteristics (unit of hospitalization and isolation measures) and risk factors. Between December 2013 and February 2014, 282 contact patients were included in the study. The prevalence of VRE acquisition was 6.4% (18/282). Significant risk factors for VRE acquisition according to logistic regression analysis were; lack of isolation, hospitalization in the same hospital unit as a VRE carrier patient and lack of isolation (RR=856.8, p=0.001), hospitalization in a specific unit (RR=927.4, p=0.002), McCabe score equal to 2 (RR=5233.6, p=0.008), age (RR=1.2 by year, p=0.011), hemodialysis (RR=36.1, p=0.011), central venous catheter (RR=25.4, p=0.021) and surgery (RR=0.012, p=0.007). Antibiotic use was a significant risk factor for VRE acquisition using univariate analysis (p<10-3). The findings confirm that the factors focused on by the study (lack of isolation and dedicated unit) had a significant effect on VRE acquisition as patient associated factors. It highlights the importance of observance of the guidelines.

Combined Genotypic, Phylogenetic, and Epidemiologic Analyses of Mycobacterium tuberculosis Genetic Diversity in the Rhône Alpes Region, France.

BACKGROUND: The present work relates to identification and a deep molecular characterization of circulating Mycobacterium tuberculosis complex (MTBC) strains in the Rhône-Alpes region, France from 2000 to 2010. It aimed to provide with a first snapshot of MTBC genetic diversity in conjunction with bacterial drug resistance, type of disease and available demographic and epidemiologic characteristics over an eleven-year period, in the south-east of France. METHODS: Mycobacterium tuberculosis complex (MTBC) strains isolated in the Rhône-Alpes region, France (n = 2257, 1 isolate per patient) between 2000 and 2010 were analyzed by spoligotyping. MIRU-VNTR typing was applied on n = 1698 strains (with full results available for 974 strains). The data obtained were compared with the SITVIT2 database, followed by detailed genotyping, phylogenetic, and epidemiologic analyses in correlation with anonymized data on available demographic, and epidemiologic characteristics, and location of disease (pulmonary or extrapulmonary TB). RESULTS: The most predominant spoligotyping clusters were SIT53/T1 (n = 346, 15.3%) > SIT50/H3 (n = 166, 7.35%) > SIT42/LAM9 (n = 125, 5.5%) > SIT1/Beijing (n = 72, 3.2%) > SIT47/H1 (n = 71, 3.1%). Evolutionary-recent strains belonging to the Principal Genetic Group (PGG) 2/3, or Euro-American lineages (T, LAM, Haarlem, X, S) were predominant and represented 1768 or 78.33% of all isolates. For strains having drug resistance information (n = 1119), any drug resistance accounted for 14.83% cases vs. 1.52% for multidrug resistance (MDR); and was significantly more associated with age group 21-40 years (p-value<0.001). Extra-pulmonary TB was more common among female patients while pulmonary TB predominated among men (p-value<0.001; OR = 2.16 95%CI [1.69; 2.77]). Also, BOV and CAS lineages were significantly well represented in patients affected by extra-pulmonary TB (p-value<0.001). The origin was known for 927/2257 patients: 376 (40.6%) being French-born vs. 551 (59.4%) Foreign-born. French patients were significantly older (mean age: 58.42 yrs 95%CI [56.04; 60.80]) than Foreign-born patients (mean age: 42.38 yrs. 95%CI [40.75; 44.0]). CONCLUSION: The study underlined the importance of imported TB cases on the genetic diversity and epidemiologic characteristics of circulating MTBC strains in Rhône-Alpes region, France over a large time-period. It helps better understand intricate relationships between certain lineages and geographic origin of the patients, and pinpoints genotypic and phylogenetic specificities of prevailing MTBC strains. Lastly, it also demonstrated a slow decline in isolation of M. africanum lineage in this region between 2000 and 2010.