OBJECTIVES: High-quality diagnosis of bloodstream infections (BSI) is important for successful patient management. As knowledge on current practices of microbiological BSI diagnostics is limited, this project aimed to assess its current state in European microbiological laboratories. METHODS: We performed an online questionnaire-based cross-sectional survey comprising 34 questions on practices of microbiological BSI diagnostics. The ESCMID Study Group for Bloodstream Infections, Endocarditis and Sepsis (ESGBIES) was the primary platform to engage national coordinators who recruited laboratories within their countries. RESULTS: Responses were received from 209 laboratories in 25 European countries. Although 32.5% (68/209) of laboratories only used the classical processing of positive blood cultures (BC), two-thirds applied rapid technologies. Of laboratories that provided data, 42.2% (78/185) were able to start incubating BC in automated BC incubators around-the-clock, and only 13% (25/192) had established a 24-h service to start immediate processing of positive BC. Only 4.7% (9/190) of laboratories validated and transmitted the results of identification and antimicrobial susceptibility testing (AST) of BC pathogens to clinicians 24 h/day. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry from briefly incubated sub-cultures on solid media was the most commonly used approach to rapid pathogen identification from positive BC, and direct disc diffusion was the most common rapid AST method from positive BC. CONCLUSIONS: Laboratories have started to implement novel technologies for rapid identification and AST for positive BC. However, progress is severely compromised by limited operating hours such that current practice of BC diagnostics in Europe complies only partly with the requirements for optimal BSI management.

• Klíčová slova
• Antimicrobial susceptibility testing, Blood culture, Bloodstream infection, Identification, Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, Microbiological diagnostics, Molecular methods, Preanalytics, Rapid diagnostics, Transport,
• MeSH
• diagnostické techniky molekulární metody   MeSH
• diagnostické testy rutinní metody   MeSH
• lidé MeSH
• mikrobiologické techniky metody   MeSH
• průřezové studie MeSH
• sepse diagnóza   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Evropa MeSH

BACKGROUND: This study introduces an advanced 8-well loop-mediated isothermal amplification (LAMP) system specifically designed for the automated colorimetric detection of SARS-CoV-2. Incorporating two distinct configurations having either three light-emitting diodes (LEDs) with varying emission wavelengths per well, paired with a photodiode detector, or utilizing white LED illumination with a red, green, and blue (RGB) sensor. The colorimetric LAMP aims to provide a more accessible and rapid diagnostic tool than traditional fluorescence methods due to the system's simplicity. RESULTS: We designed, assembled, and compared two colorimetric home-assembled LAMP systems, the first one based on three LEDs, each with a different color with a photodiode, and the second one having RGB and a white LED, with traditional fluorescence-based LAMP method performed on a commercial qPCR instrument. Results demonstrated that the colorimetric RT-LAMP assays achieved critical threshold time (CT), closely matching the CT value of fluorescence-based detection accomplished by the qPCR instrument. We performed the fundamental experiment employing an identical RNA copy number of 1,570copies·μL-1, getting the CT value of (16.70 ± 0.43) min (mean ± standard deviation from 23 measurements). Then, we also performed different RNA numbers of copies between the highest and lowest RNA contents of ≈ 157,000 copies·μL-1 and ≈ 1570 copies·μL-1, respectively, getting CT values from (13.30 ± 0.04) min to (13.75 ± 0.30) min and (17.04 ± 0.02) min to (17.26 ± 0.02) min, all (mean ± standard deviation from three measurements). The colorimetric systems demonstrated rapid response and precision across varied viral loads while keeping the system simple due to the colorimetric detection method. SIGNIFICANCE AND NOVELTY: The LAMP system's rapid and precise detection capabilities underscore its potential as an effective tool for point-of-need diagnostics. It is crucial for timely responses in ongoing and future pandemic scenarios. This system enhances testing accessibility and provides a robust platform for potential adaptation to other pathogenic threats, making it a valuable asset in global health diagnostics.

• Klíčová slova
• Biosensors and actuators, Colorimetric detection, Loop-mediated isothermal amplification (LAMP), Point-of-care diagnostics, Rapid testing technologies, SARS-CoV-2 diagnostics,
• MeSH
• COVID-19 * diagnóza   virologie   MeSH
• diagnostické techniky molekulární metody   MeSH
• kolorimetrie * metody   MeSH
• lidé MeSH
• RNA virová analýza   genetika   MeSH
• SARS-CoV-2 * genetika   izolace a purifikace   MeSH
• techniky amplifikace nukleových kyselin * metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• RNA virová MeSH

BACKGROUND: Glioblastoma (GBM) is the most common and aggressive primary brain cancer. The treatment of GBM consists of a combination of surgery and subsequent oncological therapy, i.e., radiotherapy, chemotherapy, or their combination. If postoperative oncological therapy involves irradiation, magnetic resonance imaging (MRI) is used for radiotherapy treatment planning. Unfortunately, in some cases, a very early worsening (progression) or return (recurrence) of the disease is observed several weeks after the surgery and is called rapid early progression (REP). Radiotherapy planning is currently based on MRI for target volumes definitions in many radiotherapy facilities. However, patients with REP may benefit from targeting radiotherapy with other imaging modalities. The purpose of the presented clinical trial is to evaluate the utility of 11C-methionine in optimizing radiotherapy for glioblastoma patients with REP. METHODS: This study is a nonrandomized, open-label, parallel-setting, prospective, monocentric clinical trial. The main aim of this study was to refine the diagnosis in patients with GBM with REP and to optimize subsequent radiotherapy planning. Glioblastoma patients who develop REP within approximately 6 weeks after surgery will undergo 11C-methionine positron emission tomography (PET/CT) examinations. Target volumes for radiotherapy are defined using both standard planning T1-weighted contrast-enhanced MRI and PET/CT. The primary outcome is progression-free survival defined using RANO criteria and compared to a historical cohort with REP treated without PET/CT optimization of radiotherapy. DISCUSSION: PET is one of the most modern methods of molecular imaging. 11C-Methionine is an example of a radiolabelled (carbon 11) amino acid commonly used in the diagnosis of brain tumors and in the evaluation of response to treatment. Optimized radiotherapy may also have the potential to cover those regions with a high risk of subsequent progression, which would not be identified using standard-of-care MRI for radiotherapy planning. This is one of the first study focused on radiotherapy optimization for subgroup of patinets with REP. TRIAL REGISTRATION: NCT05608395, registered on 8.11.2022 in clinicaltrials.gov; EudraCT Number: 2020-000640-64, registered on 26.5.2020 in clinicaltrialsregister.eu. Protocol ID: MOU-2020-01, version 3.2, date 18.09.2020.

• Klíčová slova
• 11C-methionine, Clinical trial, Glioblastoma, Positron emission tomography, Radiopharmaceutical, Radiotherapy, Rapid early progression,
• MeSH
• dospělí MeSH
• glioblastom * diagnostické zobrazování   terapie   diagnóza   radioterapie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• magnetická rezonanční tomografie metody   MeSH
• methionin * MeSH
• nádory mozku * diagnostické zobrazování   terapie   radioterapie   diagnóza   MeSH
• PET/CT metody   MeSH
• plánování radioterapie pomocí počítače metody   MeSH
• progrese nemoci * MeSH
• prospektivní studie MeSH
• radiofarmaka terapeutické užití   MeSH
• radioizotopy uhlíku 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
• klinické zkoušky MeSH
• Názvy látek
• methionin * MeSH
• radiofarmaka MeSH
• radioizotopy uhlíku MeSH

• MeSH
• radiografie * MeSH
• Publikační typ
• časopisecké články MeSH

This review aims to explore the role of professional diagnostic rapid testing of acute respiratory infections (ARIs), especially COVID-19 and influenza, ensuring proper disease management and treatment in Europe, and particularly in Czech Republic, Poland, and Romania. The paper was constructed based on a review of scientific evidence and national and international policies and recommendations, as well as a process of validation by four experts. The development of new testing technologies, treatment options, and increased awareness of the negative multidimensional impact of ARI profiles transformed differential diagnosis into a tangible and desirable reality. This review covers the following topics: (1) the multidimensional impact of ARIs, (2) ARI rapid diagnostic testing platforms and their value, (3) the policy landscape, (4) challenges and barriers to implementation, and (5) a set of recommendations illustrating a path forward. The findings indicate that rapid diagnostic testing, including at the point of care (POC), can have a positive impact on case management, antimicrobial and antibiotic stewardship, epidemiological surveillance, and decision making. Integrating this strategy will require the commitment of governments and the international and academic communities, especially as we identified room for improvement in the access and expansion of POC rapid testing in the focus countries and the inclusion of rapid testing in relevant policies.

• Klíčová slova
• COVID-19, Europe, acute respiratory infections, diagnostic testing, differential diagnosis, health policy, influenza, point-of-care testing, rapid testing,
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Aim: Finding rapid, reliable diagnostic methods is a big challenge in clinical microbiology. Raman spectroscopy is an optical method used for multiple applications in scientific fields including microbiology. This work reports its potential in identifying biofilm positive strains of Candida parapsilosis and Staphylococcus epidermidis. Materials & methods: We tested 54 S. epidermidis strains (23 biofilm positive, 31 negative) and 51 C. parapsilosis strains (27 biofilm positive, 24 negative) from colonies on Mueller-Hinton agar plates, using Raman spectroscopy. Results: The accuracy was 98.9% for C. parapsilosis and 96.1% for S. epidermidis. Conclusion: The method showed great potential for identifying biofilm positive bacterial and yeast strains. We suggest that Raman spectroscopy might become a useful aid in clinical diagnostics.

• Klíčová slova
• Raman spectroscopy, biofilm, rapid diagnostics,
• MeSH
• biofilmy růst a vývoj   MeSH
• Candida parapsilosis metabolismus   MeSH
• diagnostické testy rutinní metody   MeSH
• lidé MeSH
• Ramanova spektroskopie metody   MeSH
• Staphylococcus epidermidis metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Lung carcinoma remains the leading cause of cancer death worldwide. The tactic to change this unfortunate rate may be a timely and rapid diagnostic, which may in many cases improve patient prognosis. In our study, we focus on the comparison of two novel methods of rapid lung carcinoma diagnostics, label-free in vivo and ex vivo Raman spectroscopy of the epithelial tissue, and assess their feasibility in clinical practice. As these techniques are sensitive not only to the basic molecular composition of the analyzed sample but also to the secondary structure of large biomolecules, such as tissue proteins, they represent suitable candidate methods for epithelial cancer diagnostics. During routine bronchoscopy, we collected 78 in vivo Raman spectra of normal and cancerous lung tissue and 37 samples of endobronchial pathologies, which were subsequently analyzed ex vivo. Using machine learning techniques, namely principal component analysis (PCA) and support vector machines (SVM), we were able to reach 87.2% (95% CI, 79.8-94.6%) and 100.0% (95% CI, 92.1-100.0%) of diagnostic accuracy for in vivo and ex vivo setup, respectively. Although the ex vivo approach provided superior results, the rapidity of in vivo Raman spectroscopy might become unmatchable in the acceleration of the diagnostic process.

• Klíčová slova
• Endoscopy, Ex vivo diagnostics, In vivo diagnostics, Lung cancer, Machine learning, Optical biopsy, Raman spectroscopy,
• MeSH
• analýza hlavních komponent * MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory plic * diagnóza   patologie   MeSH
• Ramanova spektroskopie * metody   MeSH
• senioři MeSH
• support vector machine MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Purified α1 proteinase inhibitor (A1PI) slowed emphysema progression in patients with severe α1 antitrypsin deficiency in a randomised controlled trial (RAPID-RCT), which was followed by an open-label extension trial (RAPID-OLE). The aim was to investigate the prolonged treatment effect of A1PI on the progression of emphysema as assessed by the loss of lung density in relation to RAPID-RCT. METHODS: Patients who had received either A1PI treatment (Zemaira or Respreeza; early-start group) or placebo (delayed-start group) in the RAPID-RCT trial were included in this 2-year open-label extension trial (RAPID-OLE). Patients from 22 hospitals in 11 countries outside of the USA received 60 mg/kg per week A1PI. The primary endpoint was annual rate of adjusted 15th percentile lung density loss measured using CT in the intention-to-treat population with a mixed-effects regression model. This trial is registered with ClinicalTrials.gov, number NCT00670007. FINDINGS: Between March 1, 2006, and Oct 13, 2010, 140 patients from RAPID-RCT entered RAPID-OLE: 76 from the early-start group and 64 from the delayed-start group. Between day 1 and month 24 (RAPID-RCT), the rate of lung density loss in RAPID-OLE patients was lower in the early-start group (-1·51 g/L per year [SE 0·25] at total lung capacity [TLC]; -1·55 g/L per year [0·24] at TLC plus functional residual capacity [FRC]; and -1·60 g/L per year [0·26] at FRC) than in the delayed-start group (-2·26 g/L per year [0·27] at TLC; -2·16 g/L per year [0·26] at TLC plus FRC, and -2·05 g/L per year [0·28] at FRC). Between months 24 and 48, the rate of lung density loss was reduced in delayed-start patients (from -2·26 g/L per year to -1·26 g/L per year), but no significant difference was seen in the rate in early-start patients during this time period (-1·51 g/L per year to -1·63 g/L per year), thus in early-start patients the efficacy was sustained to month 48. INTERPRETATION: RAPID-OLE supports the continued efficacy of A1PI in slowing disease progression during 4 years of treatment. Lost lung density was never recovered, highlighting the importance of early intervention with A1PI treatment. FUNDING: CSL Behring.

• MeSH
• alfa-1-antitrypsin aplikace a dávkování   MeSH
• celková kapacita plic MeSH
• deficit alfa1-antitrypsinu komplikace   patologie   MeSH
• dospělí MeSH
• inhibitory serinových proteinas aplikace a dávkování   MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• plíce patologie   patofyziologie   MeSH
• plicní emfyzém vrozené   farmakoterapie   patologie   MeSH
• progrese nemoci MeSH
• regresní analýza MeSH
• respirační funkční testy MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• randomizované kontrolované studie MeSH
• Názvy látek
• alfa-1-antitrypsin MeSH
• inhibitory serinových proteinas MeSH

Xanthomonas euvesicatoria and X. vesicatoria are two economically important causal agents of bacterial spot (BS) of tomato and pepper. Management of BS in the field requires rapid and accurate detection. Therefore, this work aimed to develop a pipeline to design a simple, fast, and reliable assay for the detection of X. euvesicatoria and X. vesicatoria by loop-mediated isothermal amplification. In total, 109 publicly available whole genomic sequences of 24 different species of bacterial pathogens were used to design primers that would amplify the DNA of the two target species. Laboratory testing of the assay was performed on pure bacterial cultures and artificially infected plants, and amplification was conducted with both a sophisticated laboratory instrument and a simple mobile platform. The testing of the assay confirmed its specificity with a sensitivity reaching 1 pg µl-1 for both pathogens with an assay duration of 40 min on a mobile detection platform. Our diagnostics development pipeline enables the easy and fast design of a reliable detection assay in the genomics age. By validating the pipeline with X. euvesicatoria and X. vesicatoria pathogens, we have simultaneously developed an assay with high specificity, sensitivity, and speed, which will allow it to be deployed, contributing to successful management of BS.

• Klíčová slova
• LAMP, LAMP pipeline, Xanthomonas euvesicatoria, Xanthomonas vesicatoria, bacterial spot of tomato and pepper, diagnostic assay, diagnostics pipeline,
• MeSH
• diagnostické techniky molekulární MeSH
• Solanum lycopersicum * MeSH
• techniky amplifikace nukleových kyselin MeSH
• Xanthomonas * genetika   MeSH
• Publikační typ
• časopisecké články MeSH

Clinical treatment of the infections caused by various staphylococcal species differ depending on the actual cause of infection. Therefore, it is necessary to develop a fast and reliable method for identification of staphylococci. Raman spectroscopy is an optical method used in multiple scientific fields. Recent studies showed that the method has a potential for use in microbiological research, too. Our work here shows a possibility to identify staphylococci by Raman spectroscopy. We present a method that enables almost 100% successful identification of 16 of the clinically most important staphylococcal species directly from bacterial colonies grown on a Mueller-Hinton agar plate. We obtained characteristic Raman spectra of 277 staphylococcal strains belonging to 16 species from a 24-hour culture of each strain grown on the Mueller-Hinton agar plate using the Raman instrument. The results show that it is possible to distinguish among the tested species using Raman spectroscopy and therefore it has a great potential for use in routine clinical diagnostics.

• MeSH
• agar MeSH
• analýza hlavních komponent MeSH
• časové faktory MeSH
• diagnostické testy rutinní MeSH
• fluorescence MeSH
• odběr biologického vzorku MeSH
• Ramanova spektroskopie metody   MeSH
• Staphylococcus izolace a purifikace   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• agar MeSH