OBJECTIVES: Treponema pallidum subsp. pallidum (T. pallidum) is the etiological agent of syphilis, a sexually transmitted disease of global public health importance. The objective of this study was to introduce a novel in vitro protocol for isolation of T. pallidum directly from patients' clinical samples, eliminating the need for rabbit propagation. METHODS: Four oral and five genital swabs were collected from nine epidemiologically unrelated patients at two hospitals in Brno, Czech Republic. Swabs were submerged in TpCM-2 medium for transport. Samples were then placed on a 0.4 μm filters and incubated for 2.5 hours. During this period, spiral T. pallidum cells passed through the filter pores to the well containing TpCM-2 medium and rabbit feeder cells (Sf1Ep). Stable T. pallidum cultures (containing >1 × 107 treponemes) were achieved by subculturing every 7 days into fresh well. RESULTS: A successful protocol for in vitro isolation of T. pallidum was established. From the nine clinical specimens processed, six T. pallidum cultures (MU1-MU6) were derived after 14 to 112 days of cultivation. Five of these strains (MU1-MU5) belonged to SS14-like cluster and shared the same allelic profile 1.3.1. The remaining strain (MU6) was identified as a Nichols-like strain with an allelic profile 9.16.3. DISCUSSION: The introduced in vitro protocol enables isolation of T. pallidum from clinical material, including frozen samples, without the need for experimental rabbits. This method facilitates the isolation of contemporary, clinically relevant treponemal strains.

• Klíčová slova
• Contemporary isolates, In vitro isolation, Spirochetes, Syphilis, Treponema pallidum,
• MeSH
• bakteriologické techniky * metody   MeSH
• králíci MeSH
• lidé MeSH
• pohlavní orgány mikrobiologie   MeSH
• syfilis * mikrobiologie   diagnóza   MeSH
• Treponema pallidum * izolace a purifikace   genetika   klasifikace   MeSH
• ústa mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

OBJECTIVES: A number of population pharmacokinetic (popPK) models of imipenem in critically ill patients are available for dosing optimization, but they represent only a narrow range of kidney functions. This study evaluates the target attainment of on-label regimens through popPK modelling and simulation in patients across different kidney functions. METHODS: A popPK model was built based on two datasets from Switzerland (model development population, 151 patients, 322 concentrations) and externally validated on two datasets from the Czech Republic (19 patients, 111 concentrations) and Vietnam (43 patients, 85 concentrations). Monte Carlo simulations were performed to evaluate the probability of target attainment from a MIC of 0.125 mg/L to 32 mg/L. We estimated the cumulative fraction of response against Pseudomonas aeruginosa (the epidemiological cut-off value was 4 mg/L) across a broad range of Cockcroft-Gault creatinine clearance values (CLCRCG 15-130 mL/min). Targets of 40% and 100%ƒT > MIC (percentage of dosing interval estimated free concentrations above MIC) were investigated. RESULTS: Decreased kidney function estimated glomerular filtration rate by the Chronic Kidney Disease Epidemiology Collaboration 2021 (eGFRCKD-EPI <90 mL/min) was observed in 70 of 151 patients (46.4%) within the model development population, and in 11 of 19 (57.9%) and 24 of 43 (55.8%) patients in the Czech Republic and Vietnam, respectively. CLCRCG significantly influenced the imipenem clearance described by a two-compartment model. For probability of target attainment, all regimens achieved 40% ƒT > MIC2mg/L. With a 100%ƒT > MIC target, 500 mg q6h (CLCRCG 30-60 mL/min) could only cover an MIC of up to 1 mg/L, irrespective of infusion time. For cumulative fraction of response, no dosing regimen could cover susceptible P. aeruginosa for 100%ƒT > MIC. DISCUSSION: The highest on-label imipenem dosing regimens failed to attain 100% ƒT > MIC4mg/L in patients with decreased kidney function. Higher dosing may be necessary to cover MIC of 4 mg/L. Future trials should explore their efficacy, toxicity, and the utility of model-informed precision dosing in this population.

• Klíčová slova
• Critical illness, Decreased kidney function, External validation, Imipenem, Population pharmacokinetics, Simulation,
• Publikační typ
• časopisecké články MeSH

OBJECTIVES: Pseudomonas aeruginosa (PA) is a common causative pathogen of pneumonia acquired in the intensive care unit (ICU). The aim of this study was to determine the incidence of PA ICU pneumonia (PAIP) and to quantify its independent association with PA colonization at different body sites. METHODS: Adult patients on mechanical ventilation at ICU admission were prospectively enrolled across 30 European ICUs. PA colonization in the perianal area and in the lower respiratory tract was assessed within 72 hours after ICU admission and twice weekly until ICU discharge. PAIP development was evaluated daily. Competing risk models with colonization as a time-varying exposure and ICU death and discharge as competing events were fitted and adjusted for confounders to investigate the association between PA carriage and PAIP. RESULTS: A total of 1971 subjects were enrolled. The colonization prevalence with PA in the first 72 hours of ICU admission was 10.4% (179 perianal and 51 respiratory), whereas the acquisition incidence during the ICU stay was 7.0% (158 perianal and 47 respiratory). Of the 43 (1.8%) patients who developed PAIP, 11 (25.6%) were PA colonized on admission and 9 (20.9%) acquired colonization before PAIP onset. Both perianal (adjusted subdistribution hazard ratio, 4.4; 95% CI, 1.7-11.6) and respiratory colonization (adjusted subdistribution hazard ratio: 4.6, 95% CI, 1.9-11.1) were independently associated with PAIP development. DISCUSSION: PAIP incidence was higher in PA colonized vs. non-colonized patients. Colonization of both the rectum and of the respiratory tract was associated with development of PAIP. The increased risk of PA colonization for subsequent infection provides an opportunity for targeted preventive interventions.

• Klíčová slova
• Colonization, Healthcare-associated infection, ICU-acquired pneumonia, Pseudomonas aeruginosa, VAP,
• MeSH
• bakteriální pneumonie * epidemiologie   mikrobiologie   MeSH
• dospělí MeSH
• incidence MeSH
• jednotky intenzivní péče * MeSH
• lidé středního věku MeSH
• lidé MeSH
• nozokomiální pneumonie * epidemiologie   mikrobiologie   MeSH
• přenašečství * epidemiologie   mikrobiologie   MeSH
• prevalence MeSH
• prospektivní studie MeSH
• pseudomonádové infekce * epidemiologie   mikrobiologie   MeSH
• Pseudomonas aeruginosa * izolace a purifikace   MeSH
• rizikové faktory MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH
• Geografické názvy
• Evropa epidemiologie   MeSH

• Klíčová slova
• BPaLM availability, Bedaquline testing, MDR-TB, MDR-TB testing in EU/EAA, Pretomanid testing,
• MeSH
• antituberkulotika * terapeutické užití   farmakologie   MeSH
• lidé MeSH
• mikrobiální testy citlivosti * MeSH
• multirezistentní tuberkulóza * farmakoterapie   diagnóza   mikrobiologie   MeSH
• Mycobacterium tuberculosis účinky léků   genetika   MeSH
• rifampin * terapeutické užití   farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• dopisy MeSH
• komentáře MeSH
• Geografické názvy
• Evropa MeSH
• Názvy látek
• antituberkulotika * MeSH
• rifampin * MeSH

BACKGROUND: Faecal microbiota transplantation (FMT) is the standard treatment for patients with multiple recurrent Clostridioides difficile infection (rCDI). Recently, new commercially developed human microbiota-derived medicinal products have been evaluated and Food and Drug Administration-approved with considerable differences in terms of composition, administration, and targeted populations. OBJECTIVES: To review available data on the different microbiota-derived treatments at the stage of advanced clinical evaluation and research in rCDI in comparison with FMT. SOURCES: Phase II or III trials evaluating a microbiota-derived medicinal product to prevent rCDI. CONTENT: Two commercial microbiota-derived medicinal products are approved by the Food and Drug Administration: Rebyota (RBX2660 Ferring Pharmaceuticals, marketed in the United States) and VOWST (SER-109 -Seres Therapeutics, marketed in the United States), whereas VE303 (Vedanta Biosciences Inc) will be studied in phase III trial. RBX2660 and SER-109 are based on the processing of stools from healthy donors, whereas VE303 consists of a defined bacterial consortium originating from human stools and produced from clonal cell banks. All have proven efficacy to prevent rCDI compared with placebo in patients considered at high risk of recurrence. However, the heterogeneity of the inclusion criteria, and the time between each episode and CDI diagnostics makes direct comparison between trials difficult. The differences regarding the risk of recurrence between the treatment and placebo arms were lower than previously described for FMT (FMT: Δ = 50.5%; RBX2660-phase III: Δ = 13.1%; SER-109-phase III: Δ = 28%; high-dose VE303-phase-II: Δ = 31.7%). All treatments presented a good overall safety profile with mainly mild gastrointestinal symptoms. IMPLICATIONS: Stool-derived products and bacterial consortia need to be clearly distinguished in terms of product characterization and their associated risks with specific long-term post-marketing evaluation similar to registries used for FMT. Their place in the therapeutic strategy for patients with rCDI requires further studies to determine the most appropriate patient population and administration route to prevent rCDI.

• Klíčová slova
• Clostridioides difficile infection, Faecal microbiota transplantation, Live biotherapeutic products, Microbiota-derived therapy, Recurrent Clostridioides difficile infection,
• MeSH
• Clostridioides difficile * MeSH
• fekální transplantace MeSH
• klostridiové infekce * mikrobiologie   MeSH
• lidé MeSH
• mikrobiota * MeSH
• recidiva MeSH
• výsledek terapie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

OBJECTIVES: To evaluate the efficacy and tolerability of a single dose of oral cefixime 800 mg plus oral doxycycline 100 mg twice a day for 7 days, compared with a recommended single dose of ceftriaxone plus single dose of oral azithromycin, for treatment of uncomplicated urogenital, rectal, or pharyngeal gonorrhoea. METHODS: A noninferiority, open-label, multicentre randomized controlled trial was conducted in Prague, Czech Republic. Some 161 patients, 18-65 years of age diagnosed with uncomplicated urogenital, rectal, or pharyngeal gonorrhoea by nucleic acid amplification test (NAAT) were randomized to treatment with single dose of cefixime 800 mg plus doxycycline 100 mg twice a day for 1 week or a single dose of ceftriaxone 1 g intramuscularly plus single dose of azithromycin 2 g. The primary outcome was the number of participants with negative culture and NAAT at 1 week and 3 weeks, respectively, after treatment initiation. RESULTS: In all, 161 patients were randomized and 152 were included in per-protocol analyses. All 76 (100%; 95% CI, 0.95-1.00) patients treated with ceftriaxone plus azithromycin achieved negative cultures and NAAT after treatment. In the cefixime plus doxycycline arm at week 1, culture was negative in all 76 (100%) patients; at week 3, culture was negative in 70 of the 76 patients (92%; 95% CI, 0.84-0.97) and NAAT negative in 66 of the 76 patients (87%; 95% CI, 0.77-0.94). At week 3, culture and NAAT were negative in 65 of the 76 patients (86%; 95% CI, 0.76-0.93). Per-protocol risk difference was 14.5%; 95% CI, 6.56-22.38. All treatment failures observed in the cefixime arm were pharyngeal gonorrhoea cases. DISCUSSION: The combination of cefixime and doxycycline did not achieve noninferiority to ceftriaxone and azithromycin for treatment of gonorrhoea when including pharyngeal gonorrhoea. It did, however, show high efficacy for urogenital and rectal gonorrhoea.

• Klíčová slova
• Azithromycin, Cefixime, Ceftriaxone, Doxycycline, Neisseria gonorrhoeae, Treatment,
• MeSH
• antibakteriální látky terapeutické užití   MeSH
• azithromycin terapeutické užití   MeSH
• cefixim terapeutické užití   MeSH
• ceftriaxon * MeSH
• dospělí MeSH
• doxycyklin terapeutické užití   MeSH
• gonorea * farmakoterapie   mikrobiologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• Neisseria gonorrhoeae MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• hodnocení ekvivalence MeSH
• multicentrická studie MeSH
• randomizované kontrolované studie MeSH
• Názvy látek
• antibakteriální látky MeSH
• azithromycin MeSH
• cefixim MeSH
• ceftriaxon * MeSH
• doxycyklin MeSH

• Klíčová slova
• Clostridioides difficile infection, Mortality, Patient-relevant outcomes, Recurrence, Sustained cure,
• MeSH
• antibakteriální látky * terapeutické užití   MeSH
• klostridiové infekce * farmakoterapie   MeSH
• lidé MeSH
• recidiva MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• dopisy MeSH
• komentáře MeSH
• Názvy látek
• antibakteriální látky * MeSH

• Klíčová slova
• CDI, Clostridioides difficile, Infection, PPI, Patient and public involvement, Research,
• Publikační typ
• časopisecké články MeSH

• Klíčová slova
• Hand hygiene, Infectivity, Paediatric, Palmar hand swab, Respiratory virus, Smear infection,
• MeSH
• COVID-19 * diagnóza   MeSH
• dítě hospitalizované MeSH
• dítě MeSH
• lidé MeSH
• ruka MeSH
• SARS-CoV-2 MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• Publikační typ
• dopisy MeSH

OBJECTIVES: Matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) is a widely used method for bacterial species identification. Incomplete databases and mass spectral quality (MSQ) still represent major challenges. Important proxies for MSQ are the number of detected marker masses, reproducibility, and measurement precision. We aimed to assess MSQs across diagnostic laboratories and the potential of simple workflow adaptations to improve it. METHODS: For baseline MSQ assessment, 47 diverse bacterial strains, which are challenging to identify by MALDI-TOF MS, were routinely measured in 36 laboratories from 12 countries, and well-defined MSQ features were used. After an intervention consisting of detailed reported feedback and instructions on how to acquire MALDI-TOF mass spectra, measurements were repeated and MSQs were compared. RESULTS: At baseline, we observed heterogeneous MSQ between the devices, considering the median number of marker masses detected (range = [2-25]), reproducibility between technical replicates (range = [55%-86%]), and measurement error (range = [147 parts per million (ppm)-588 ppm]). As a general trend, the spectral quality was improved after the intervention for devices, which yielded low MSQs in the baseline assessment as follows: for four out of five devices with a high measurement error, the measurement precision was improved (p-values <0.001, paired Wilcoxon test); for six out of ten devices, which detected a low number of marker masses, the number of detected marker masses increased (p-values <0.001, paired Wilcoxon test). DISCUSSION: We have identified simple workflow adaptations, which, to some extent, improve MSQ of poorly performing devices and should be considered by laboratories yielding a low MSQ. Improving MALDI-TOF MSQ in routine diagnostics is essential for increasing the resolution of bacterial identification by MALDI-TOF MS, which is dependent on the reproducible detection of marker masses. The heterogeneity identified in this external quality assessment (EQA) requires further study.

• Klíčová slova
• Bacterial species identification, Diagnostic performance external quality assessment, MALDI-TOF MS, Quality control, Standardisation,
• MeSH
• Bacteria * MeSH
• laboratoře * MeSH
• lidé MeSH
• průběh práce MeSH
• reprodukovatelnost výsledků MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH