RESUMEN
In clinical bacteriology laboratories, reading and processing of sterile plates remain a significant part of the routine workload (30%-40% of the plates). Here, an algorithm was developed for bacterial growth detection starting with any type of specimens and using the most common media in bacteriology. The growth prediction performance of the algorithm for automatic processing of sterile plates was evaluated not only at 18-24 h and 48 h but also at earlier timepoints toward the development of an early growth monitoring system. A total of 3,844 plates inoculated with representative clinical specimens were used. The plates were imaged 15 times, and two different microbiologists read the images randomly and independently, creating 99,944 human ground truths. The algorithm was able, at 48 h, to discriminate growth from no growth with a sensitivity of 99.80% (five false-negative [FN] plates out of 3,844) and a specificity of 91.97%. At 24 h, sensitivity and specificity reached 99.08% and 93.37%, respectively. Interestingly, during human truth reading, growth was reported as early as 4 h, while at 6 h, half of the positive plates were already showing some growth. In this context, automated early growth monitoring in case of normally sterile samples is envisioned to provide added value to the microbiologists, enabling them to prioritize reading and to communicate early detection of bacterial growth to the clinicians.
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Inteligencia Artificial , Bacterias , Sensibilidad y Especificidad , Humanos , Bacterias/crecimiento & desarrollo , Bacterias/aislamiento & purificación , Bacterias/clasificación , Algoritmos , Técnicas Bacteriológicas/métodos , Procesamiento de Imagen Asistido por Computador/métodos , Infecciones Bacterianas/diagnóstico , Infecciones Bacterianas/microbiología , Bacteriología , Automatización de Laboratorios/métodos , Medios de Cultivo/químicaRESUMEN
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific antibody responses to the spike (S) protein monomer, S protein native trimeric form, or the nucleocapsid (N) proteins were evaluated in cohorts of individuals with acute infection (n = 93) and in individuals enrolled in a postinfection seroprevalence population study (n = 578) in Switzerland. Commercial assays specific for the S1 monomer, for the N protein, or within a newly developed Luminex assay using the S protein trimer were found to be equally sensitive in antibody detection in the acute-infection-phase samples. Interestingly, compared to anti-S antibody responses, those against the N protein appear to wane in the postinfection cohort. Seroprevalence in a "positive patient contacts" group (n = 177) was underestimated by N protein assays by 10.9 to 32.2%, while the "randomly selected" general population group (n = 311) was reduced by up to 45% relative to the S protein assays. The overall reduction in seroprevalence targeting only anti-N antibodies for the total cohort ranged from 9.4 to 31%. Of note, the use of the S protein in its native trimer form was significantly more sensitive compared to monomeric S proteins. These results indicate that the assessment of anti-S IgG antibody responses against the native trimeric S protein should be implemented to estimate SARS-CoV-2 infections in population-based seroprevalence studies.IMPORTANCE In the present study, we have determined SARS-CoV-2-specific antibody responses in sera of acute and postinfection phase subjects. Our results indicate that antibody responses against viral S and N proteins were equally sensitive in the acute phase of infection, but that responses against N appear to wane in the postinfection phase where those against the S protein persist over time. The most sensitive serological assay in both acute and postinfection phases used the native S protein trimer as the binding antigen, which has significantly greater conformational epitopes for antibody binding compared to the S1 monomer protein used in other assays. We believe these results are extremely important in order to generate correct estimates of SARS-CoV-2 infections in the general population. Furthermore, the assessment of antibody responses against the trimeric S protein will be critical to evaluate the durability of the antibody response and for the characterization of a vaccine-induced antibody response.
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Anticuerpos Antivirales/sangre , COVID-19/inmunología , Proteínas de la Nucleocápside de Coronavirus/inmunología , SARS-CoV-2/inmunología , Glicoproteína de la Espiga del Coronavirus/inmunología , COVID-19/sangre , COVID-19/epidemiología , Femenino , Humanos , Inmunoensayo , Inmunoglobulina A/sangre , Inmunoglobulina G/sangre , Masculino , Fosfoproteínas/inmunología , Multimerización de Proteína , Sensibilidad y Especificidad , Estudios Seroepidemiológicos , Glicoproteína de la Espiga del Coronavirus/química , Suiza/epidemiología , Factores de TiempoRESUMEN
BACKGROUND: Although it has been 30 years since the first automation systems were introduced in the microbiology laboratory, total laboratory automation (TLA) has only recently been recognized as a valuable component of the laboratory. A growing number of publications illustrate the potential impact of automation. TLA can improve standardization, increase laboratory efficiency, increase workplace safety, and reduce long-term costs. CONTENT: This review provides a preview of the current state of automation in clinical microbiology and covers the main developments during the last years. We describe the available hardware systems (that range from single function devices to multifunction workstations) and the challenging alterations on workflow and organization of the laboratory that have to be implemented to optimize automation. SUMMARY: Despite the many advantages in efficiency, productivity, and timeliness that automation offers, it is not without new and unique challenges. For every advantage that laboratory automation provides, there are similar challenges that a laboratory must face. Change management strategies should be used to lead to a successful implementation. TLA represents, moreover, a substantial initial investment. Nevertheless, if properly approached, there are a number of important benefits that can be achieved through implementation of automation in the clinical microbiology laboratory. Future developments in the field of automation will likely focus on image analysis and artificial intelligence improvements. Patient care, however, should remain the epicenter of all future directions and there will always be a need for clinical microbiology expertise to interpret the complex clinical and laboratory information.
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Automatización de Laboratorios , Servicios de Laboratorio Clínico , Inteligencia Artificial , Automatización , Humanos , Laboratorios , Flujo de TrabajoRESUMEN
Campylobacter genus encompasses many species, among which C. jejuni, C. coli and C. fetus are the main human pathogens. C. jejuni/coli frequently cause self-limited enteritis in immunocompetent hosts and are seldomly associated with bacteriemia. C. fetus is less common as a human pathogen. It is rarely identified in fecal samples but can sometimes be isolated in blood samples from patients with comorbidities or immunosuppression. Campylobacter fetus bacteriemia is remarkable since it is associated with endovascular and deep-seated infections.
Le genre Campylobacter comprend plusieurs espèces pathogènes pour l'homme, en particulier C. jejuni, C. coli et C. fetus. C. jejuni et C. coli sont responsables d'entérites généralement spontanément résolutives chez l'individu sain, et peu fréquemment associées à des bactériémies. C. fetus est un pathogène méconnu, rarement identifié dans les échantillons fécaux mais parfois retrouvé dans des hémocultures, en particulier chez des patients présentant des comorbidités ou immunosupprimés. La bactériémie à C. fetus se distingue par son association avec des infections endovasculaires et des foyers infectieux profonds sans symptomatologie digestive.
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Infecciones por Campylobacter , Campylobacter , Enteritis , Infecciones por Campylobacter/diagnóstico , Infecciones por Campylobacter/epidemiología , Campylobacter fetus , Heces , HumanosRESUMEN
In clinical practice, fecal microbiota transplantation (FMT) has been established as an unparalleled therapy to date for multiple recurrent Clostridioides difficile infections (CDI). The implementation of the FMT in practice requires a significant investment to meet legal, security and financial requirements. Research on the microbiota is booming and multiple investigations on FMT in indications other than CDI are ongoing.
En pratique clinique, la transplantation de microbiote fécal (TMF) s'est établie comme une thérapie sans équivalent à ce jour pour les infections à Clostridioides difficile (C. difficile) multirécidivantes. La mise en place de la TMF en pratique demande un investissement important pour répondre aux exigences légales, sécuritaires et financières. La recherche sur le microbiote est en plein essor et de multiples recherches sur la TMF dans d'autres indications que pour l'infection à C. difficile sont en cours.
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Clostridioides difficile , Infecciones por Clostridium , Microbiota , Infecciones por Clostridium/terapia , Trasplante de Microbiota Fecal , Humanos , Recurrencia , Resultado del TratamientoRESUMEN
The 1,3-beta-d-glucan (BDG) test is used for the diagnosis of invasive candidiasis (IC) in intensive care units (ICUs). However, its utility for patient management is unclear. This study assessed the impact of BDG test results on therapeutic decisions. This was a single-center observational study conducted in an ICU over two 6-month periods. All BDG test requests for the diagnosis of IC were analyzed. Before the second period, the ICU physicians received a pocket card instruction (algorithm) for targeted BDG testing in high-risk patients. The performance of the BDG test for IC diagnosis was assessed, as well as its impact on antifungal (AF) prescription. Overall, 72 patients had ≥1 BDG test, and 14 (19%) patients had an IC diagnosis. The BDG test results influenced therapeutic decisions in 41 (57%) cases. The impact of the BDG test was positive in 30 (73%) of them, as follows: AF abstention/interruption following a negative BDG result (n = 27), and AF initiation/continuation triggered by a positive BDG test result and subsequently confirmed IC (n = 3). In 10 (24%) cases, a positive BDG test result resulted in AF initiation/continuation with no further evidence of IC. A negative BDG result and AF abstention with subsequent IC diagnosis were observed in one case. The positive predictive value (PPV) of BDG was improved if testing was restricted to the algorithm's indications (80% versus 36%, respectively). However, adherence to the algorithm was low (26%), and no benefit of the intervention was observed. The BDG result had an impact on therapeutic decisions in more than half of the cases, which consisted mainly of safe AF interruption/abstention. Targeted BDG testing in high-risk patients improves PPV but is difficult to achieve in ICU.
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Candidiasis Invasiva , beta-Glucanos , Antifúngicos/uso terapéutico , Candidiasis Invasiva/diagnóstico , Candidiasis Invasiva/tratamiento farmacológico , Cuidados Críticos , Humanos , Unidades de Cuidados IntensivosRESUMEN
Since its emergence in December 2019, scientific knowledge about the SARS-CoV-2 virus has evolved rapidly but, due to the complexity and novelty of this infection and its political and economic stakes, much remains to be clarified. Thousands of studies have already been published and scientific research is constantly evolving. In this multitude of information, we offer an update of the knowledge currently available. A limitation of the propagation, the understanding of the functioning of the virus and its clinical manifestations, the administration of specific treatments, rapid and reliable diagnostic tools are the basis of the fight against this germ, which is still little known today.
Depuis son apparition en Décembre 2019, les connaissances scientifiques concernant le virus SARS-CoV-2 ont rapidement évolués mais, en raison de la complexité et nouveauté de cette infection et de ses enjeux politiques et économiques, encore beaucoup reste à clarifier. Des milliers d'études ont déjà été publiés et la recherche scientifique est en constante évolution. Dans cette multitude d'informations, nous proposons une mise à jour des connaissances actuellement disponibles. Une limitation de la propagation, la compréhension du fonctionnement du virus et de ses manifestations cliniques, l'administration de traitements spécifiques et des outils diagnostiques rapides et fiables, sont à la base de la lutte contre ce germe à présent encore méconnu.
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Betacoronavirus , Infecciones por Coronavirus , Pandemias , Neumonía Viral , COVID-19 , Infecciones por Coronavirus/epidemiología , Infecciones por Coronavirus/terapia , Infecciones por Coronavirus/transmisión , Humanos , Neumonía Viral/epidemiología , Neumonía Viral/terapia , Neumonía Viral/transmisión , SARS-CoV-2RESUMEN
OBJECTIVES: The best therapeutic approach for treating MRSA endocarditis remains unknown, particularly in cases of high vancomycin MICs. We report here a case of daptomycin-non-susceptible, ceftaroline-resistant and fosfomycin-resistant MRSA native left valve endocarditis that was successfully treated with valve repair and a combination of high-dose daptomycin and ceftaroline. METHODS: Antimicrobial testing of the clinical strain was performed using Etest and microdilution broth methods. Time-kill and chequerboard methodologies were used to test the activity of antibiotic combinations. RESULTS: By Etest, the MIC of vancomycin was 2 mg/L, the MIC of daptomycin was 2 mg/L, the MIC of fosfomycin was 1024 mg/L and the MIC of ceftaroline was 1.5 mg/L. At the standard inoculum (105 cfu/mL), the three combinations of daptomycin plus ceftaroline, cloxacillin or fosfomycin were synergistic and bactericidal. However, when these combinations were tested using a higher inoculum (108 cfu/mL), all combinations were synergistic, but only daptomycin plus ceftaroline had bactericidal activity. CONCLUSIONS: These results confirmed a synergistic effect between daptomycin plus ceftaroline and increased bactericidal activity against MRSA, suggesting that this combination may be effective for the treatment of invasive MRSA infection. Our experience highlights the potential clinical use of synergy testing to guide difficult treatment decisions in patients with MDR MRSA infection.
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Antibacterianos/uso terapéutico , Cefalosporinas/uso terapéutico , Daptomicina/uso terapéutico , Endocarditis/tratamiento farmacológico , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Infecciones Estafilocócicas/tratamiento farmacológico , Anciano de 80 o más Años , Infecciones Comunitarias Adquiridas/diagnóstico , Infecciones Comunitarias Adquiridas/tratamiento farmacológico , Infecciones Comunitarias Adquiridas/microbiología , Sinergismo Farmacológico , Endocarditis/diagnóstico , Endocarditis/microbiología , Fosfomicina/farmacología , Humanos , Masculino , Pruebas de Sensibilidad Microbiana , Infecciones Estafilocócicas/diagnóstico , Infecciones Estafilocócicas/microbiología , CeftarolinaRESUMEN
PURPOSE OF REVIEW: This review details the management of Pseudomonas aeruginosa infections covering both current and future treatment options that are and may be available for the clinicians. RECENT FINDINGS: Pseudomonas aeruginosa infections are a great concern in hospital-acquired infections with very limited therapeutic options. The increasing antibiotic resistance has led to a need for different treatment choices that range from the use of new antibiotics to new nonantibiotic alternative agents to kill or disarm the pathogen. SUMMARY: New molecules such as ceftolozane-tazobactam, ceftazidime-avibactam, and imipenem-relebactam have shown an adequate activity against P. aeruginosa, especially against multidrug resistance strains. Other nonantibiotic alternative treatments, such as antibodies, bacteriocins or phage therapy, have shown promising results, but future clinical studies are needed.
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Antibacterianos , Farmacorresistencia Bacteriana Múltiple , Infecciones por Pseudomonas , Pseudomonas aeruginosa/efectos de los fármacos , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Bacteriemia/tratamiento farmacológico , Bacteriemia/microbiología , Humanos , Neumonía Asociada al Ventilador/tratamiento farmacológico , Neumonía Asociada al Ventilador/microbiología , Infecciones por Pseudomonas/tratamiento farmacológico , Infecciones por Pseudomonas/microbiologíaRESUMEN
Probiotic yogurt and milk supplemented with probiotics have been investigated for their role in 'low-grade' inflammation but evidence for their efficacy is inconclusive. This study explores the impact of probiotic yogurt on metabolic and inflammatory biomarkers, with a parallel study of gut microbiota dynamics. The randomised cross-over study was conducted in fourteen healthy, young men to test probiotic yogurt compared with milk acidified with 2 % d-(+)-glucono-δ-lactone during a 2-week intervention (400 g/d). Fasting assessments, a high-fat meal test (HFM) and microbiota analyses were used to assess the intervention effects. Baseline assessments for the HFM were carried out after a run-in during which normal milk was provided. No significant differences in the inflammatory response to the HFM were observed after probiotic yogurt compared with acidified milk intake; however, both products were associated with significant reductions in the inflammatory response to the HFM compared with the baseline tests (assessed by IL6, TNFα and chemokine ligand 5) (P<0·001). These observations were accompanied by significant changes in microbiota taxa, including decreased abundance of Bilophila wadsworthia after acidified milk (log 2-fold-change (FC)=-1·5, P adj=0·05) and probiotic yogurt intake (FC=-1·3, P adj=0·03), increased abundance of Bifidobacterium species after acidified milk intake (FC=1·4, P adj=0·04) and detection of Lactobacillus delbrueckii spp. bulgaricus (FC=7·0, P adj<0·01) and Streptococcus salivarius spp. thermophilus (FC=6·0, P adj<0·01) after probiotic yogurt intake. Probiotic yogurt and acidified milk similarly reduce postprandial inflammation that is associated with a HFM while inducing distinct changes in the gut microbiota of healthy men. These observations could be relevant for dietary treatments that target 'low-grade' inflammation.
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Tracto Gastrointestinal/microbiología , Leche/química , Probióticos , Yogur , Adulto , Animales , Grasas de la Dieta , Método Doble Ciego , Humanos , Masculino , Comidas , Microbiota/fisiología , Periodo Posprandial , Adulto JovenRESUMEN
Until recently, the search for enteropathogens causing travellers' diarrhea was based on stool culture (Campylobacter spp., Salmonella spp. and Shigella spp.), direct microscopy with (Cryptosporidium spp.) or without specific staining (Giardia lamblia, Entamoeba histolytica) or specific antigen detection (Giardia lamblia, Entamoeba histolytica). Molecular analyses are progressively replacing traditional diagnostic methods but their clinical usefulness remains to be better defined. This article attempts to describe the advantages and disadvantages of these new molecular methods and to illustrate situations where they could be useful using clinical cases frequently encountered in the practice of travel medicine.
Jusqu'à présent, la recherche d'entéropathogènes à l'origine de diarrhées au retour de voyage se basait essentiellement sur la culture bactérienne de selles (Campylobacter spp., Salmonella spp. et Shigella spp.), la microscopie directe sans(Giardia lamblia, Entamoeba histolytica) ou avec coloration spéciale (Cryptosporidium spp.) et la recherche d'antigènes spécifiques (Giardia lamblia, Entamoeba histolytica). Désormais, les analyses moléculaires tendent à supplanter les techniques traditionnelles mais l'utilité clinique de la PCR par rapport aux examens conventionnels doit être mieux définie. Cet article cherche à décrire les avantages et les limitations de ces nouvelles méthodes moléculaires et à illustrer des situations dans lesquelles leur utilisation pourrait être indiquée à la lumière de cas cliniques fréquemment rencontrés dans la pratique de la médecine des voyages.
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Diarrea/diagnóstico , Microbioma Gastrointestinal/genética , Reacción en Cadena de la Polimerasa Multiplex , Diarrea/microbiología , Diarrea/virología , Tracto Gastrointestinal/microbiología , Tracto Gastrointestinal/virología , Humanos , Reacción en Cadena de la Polimerasa Multiplex/normas , Valor Predictivo de las Pruebas , Sensibilidad y Especificidad , Viaje , Medicina del Viajero/métodosRESUMEN
The quality of sample inoculation is critical for achieving an optimal yield of discrete colonies in both monomicrobial and polymicrobial samples to perform identification and antibiotic susceptibility testing. Consequently, we compared the performance between the InoqulA (BD Kiestra), the WASP (Copan), and manual inoculation methods. Defined mono- and polymicrobial samples of 4 bacterial species and cloudy urine specimens were inoculated on chromogenic agar by the InoqulA, the WASP, and manual methods. Images taken with ImagA (BD Kiestra) were analyzed with the VisionLab version 3.43 image analysis software to assess the quality of growth and to prevent subjective interpretation of the data. A 3- to 10-fold higher yield of discrete colonies was observed following automated inoculation with both the InoqulA and WASP systems than that with manual inoculation. The difference in performance between automated and manual inoculation was mainly observed at concentrations of >10(6) bacteria/ml. Inoculation with the InoqulA system allowed us to obtain significantly more discrete colonies than the WASP system at concentrations of >10(7) bacteria/ml. However, the level of difference observed was bacterial species dependent. Discrete colonies of bacteria present in 100- to 1,000-fold lower concentrations than the most concentrated populations in defined polymicrobial samples were not reproducibly recovered, even with the automated systems. The analysis of cloudy urine specimens showed that InoqulA inoculation provided a statistically significantly higher number of discrete colonies than that with WASP and manual inoculation. Consequently, the automated InoqulA inoculation greatly decreased the requirement for bacterial subculture and thus resulted in a significant reduction in the time to results, laboratory workload, and laboratory costs.
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Bacterias/aislamiento & purificación , Técnicas Bacteriológicas/métodos , Manejo de Especímenes/métodos , Automatización de Laboratorios/métodos , Infecciones Bacterianas/diagnóstico , Humanos , Procesamiento de Imagen Asistido por Computador , Factores de TiempoRESUMEN
Waddlia chondrophila is an intracellular bacterium suspected to cause human and bovine abortion. We confirmed an association between antibodies against W. chondrophila and human miscarriage and identified this organism in placenta or genital tract of women who had had miscarriages. These results suggest a possible role of W. chondrophila infection in miscarriage.
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Aborto Espontáneo/epidemiología , Aborto Espontáneo/etiología , Infecciones por Chlamydia/complicaciones , Chlamydia/clasificación , Placenta/microbiología , Adulto , Estudios de Casos y Controles , Chlamydia/genética , Chlamydia/aislamiento & purificación , Infecciones por Chlamydia/diagnóstico , Femenino , Humanos , Placenta/patología , Embarazo , Factores de Riesgo , Estudios SeroepidemiológicosRESUMEN
Amoebae are unicellular protozoan present worldwide in several environments mainly feeding on bacteria. Some of them, the amoebae-resistant bacteria (ARBs), have evolved mechanisms to survive and replicate inside amoebal species. These mainly include legionella, mycobacteria and Chlamydia-related bacteria. Amoebae can provide a replicative niche, can act as reservoir for bacteria whereas the cystic form can protect the internalized bacteria. Moreover, the amoebae represent a Trojan horse for ARBs to infect animals. The long interaction between amoebae and bacteria has likely selected for bacterial virulence traits leading to the adaptation towards an intracellular lifestyle, and some ARBs have acquired the ability to infect mammals. This review intends to highlight the important uses of amoebae in several fields in microbiology by describing the main tools developed using amoebal cells. First, amoebae such as Acanthamoeba are used to isolate and discover new intracellular bacterial species by two main techniques: the amoebal co-culture and the amoebal enrichment. In the second part, taking Waddlia chondrophila as example, we summarize some important recent applications of amoebae to discover new bacterial virulence factors, in particular thanks to the amoebal plaque assay. Finally, the genetically tractable Dictyostelium discoideum is used as a model organism to study host-pathogen interactions, in particular with the development of several approaches to manipulate its genome that allowed the creation of a wide range of mutated strains largely shared within the Dictyostelium community.
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Amoeba/microbiología , Bacterias/crecimiento & desarrollo , Interacciones Huésped-Patógeno , Técnicas Microbiológicas/métodos , Factores de Virulencia/análisis , Bacterias/aislamiento & purificaciónRESUMEN
Total laboratory automation (TLA) is a valuable component of microbiology laboratories and a growing number of publications suggest the potential impact of automation in terms of analysis standardization, streaking quality, and the turnaround time (TAT). The aim of this project was to perform a detailed investigation of the impact of TLA on the workflow of commonly treated specimens such as urine. This is a retrospective observational study comparing two time periods (pre TLA versus post TLA) for urine specimen culture processing. A total of 35,864 urine specimens were plated during the pre-TLA period and 47,283 were plated during the post-TLA period. The median time from streaking to identification decreased from 22.3 h pre TLA to 21.4 h post TLA (p < 0.001), and the median time from streaking to final validation of the report decreased from 24.3 h pre TLA to 23 h post TLA (p < 0.001). Further analysis revealed that the observed differences in TAT were mainly driven by the contaminated and positive samples. Our findings demonstrate that TLA has the potential to decrease turnaround times of samples in a laboratory. Nevertheless, changes in laboratory workflow (such as extended opening hours for plate reading and antibiotic susceptibility testing or decreased incubation times) might further maximize the efficiency of TLA and optimize TATs.
RESUMEN
Members of the Chlamydiales order all share a biphasic lifecycle alternating between small infectious particles, the elementary bodies (EBs) and larger intracellular forms able to replicate, the reticulate bodies. Whereas the classical Chlamydia usually harbours round-shaped EBs, some members of the Chlamydia-related families display crescent and star-shaped morphologies by electron microscopy. To determine the impact of fixative methods on the shape of the bacterial cells, different buffer and fixative combinations were tested on purified EBs of Criblamydia sequanensis, Estrella lausannensis, Parachlamydia acanthamoebae, and Waddlia chondrophila. A linear discriminant analysis was performed on particle metrics extracted from electron microscopy images to recognize crescent, round, star and intermediary forms. Depending on the buffer and fixatives used, a mixture of alternative shapes were observed in varying proportions with stars and crescents being more frequent in C. sequanensis and P. acanthamoebae, respectively. No tested buffer and chemical fixative preserved ideally the round shape of a majority of bacteria and other methods such as deep-freezing and cryofixation should be applied. Although crescent and star shapes could represent a fixation artifact, they certainly point towards a diverse composition and organization of membrane proteins or intracellular structures rather than being a distinct developmental stage.
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Chlamydiales/clasificación , Chlamydiales/ultraestructura , Coloración y Etiquetado/métodosRESUMEN
Vaccine-induced protection against tick-borne encephalitis virus (TBEV) is mediated by antibodies to the viral particle/envelope protein. The detection of non-structural protein 1 (NS1) specific antibodies has been suggested as a marker indicative of natural infections. However, recent work has shown that TBEV vaccines contain traces of NS1, and immunization of mice induced low amounts of NS1-specific antibodies. In this study, we investigated if vaccination induces TBEV NS1-specific antibodies in humans. Healthy army members (n = 898) were asked to fill in a questionnaire relating to flavivirus vaccination or infection, and blood samples were collected. In addition, samples of 71 suspected acute TBE cases were included. All samples were screened for the presence of TBEV NS1-specific IgG antibodies using an in-house developed ELISA. Antibodies were quantified as percent positivity in reference to a positive control. For qualitative evaluation, cut-off for positivity was defined based on the mean OD of the lower 95% of the vaccinated individuals + 3 SD. We found significantly higher NS1-specific IgG antibody titers (i.e., quantitative evaluation) in individuals having received 2, 3, or 4 or more vaccine doses than in non-vaccinated individuals. Similarly, the percentage of individuals with a positive test result (i.e., qualitative evaluation) was higher in individuals vaccinated against tick-borne encephalitis than in unvaccinated study participants. Although NS1-specific IgG titers remained at a relatively low level when compared to TBE patients, a clear distinction was not always possible. Establishing a clear cut-off point in detection systems is critical for NS1-specific antibodies to serve as a marker for distinguishing the immune response after vaccination and infection.
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Virus de la Encefalitis Transmitidos por Garrapatas , Encefalitis Transmitida por Garrapatas , Infecciones por Flavivirus , Vacunas Virales , Humanos , Anticuerpos Antivirales , Formación de Anticuerpos , Encefalitis Transmitida por Garrapatas/prevención & control , Inmunoglobulina G , VacunaciónRESUMEN
OBJECTIVES: Rapid antibiotic susceptibility testing (AST) for positive blood cultures can improve patient clinical outcomes if the time to an effective antimicrobial therapy is shortened. In this study, we tested the Quantamatrix dRAST system (QMAC-dRAST), a rapid AST system based on time-lapse microscopic imagery of bacterial colony formation in agarose. METHODS: Evaluation of the QMAC-dRAST was performed from 250 monobacterial blood cultures including 130 Enterobacterales, 20 non-fermentative Gram-negative bacteria, 69 staphylococci and 31 enterococci. Blood cultures were recovered from anonymous patients or from spiking experiments to enrich our study with bacterial species and resistant strains. Categorical agreement (CA), minor errors (me), major errors (ME) and very major errors (VME) were calculated in comparison to the results obtained from the BD Phoenix™ M50. Discrepancies between the Phoenix™ M50 and QMAC-dRAST results were investigated using the gradient strip method. The repeatability and reproducibility performance of the QMAC-dRAST was assessed for 16 strains, each strain being tested five times from a spiked blood culture. RESULTS: The overall CAs for Enterobacterales, non-fermentative Gram-negative bacteria, staphylococci and enterococci were 95.1%, 91.2%, 93.4% and 94.5%, respectively. The VME percentage was below 4% for all the groups except for staphylococci, which showed a VME rate of 7%. The median time to result was 6.7 h (range: 4.7-7.9). Repeatability and reproducibility assays showed a high reliability of AST results with best and worst ratios of 98.8% and 99.6% and 95.0% and 98.3%, respectively. CONCLUSIONS: The QMAC-dRAST is a fast and reliable system to determine AST directly from monobacterial blood cultures with a major TAT reduction compared to conventional AST testing.
RESUMEN
BACKGROUND: Campylobacter spp. are a frequent cause of gastroenteritis, presenting in some patients as an acute abdominal emergency. Here we describe the distinctive clinical characteristics of these patients. METHODS: We designed a retrospective, single-centre, observational study. Children and adolescents under 18 years of age who had positive stool cultures for Campylobacter spp. during the period between June 1, 2008 and May 31, 2016 were identified from our database. Hospitalised patients with Campylobacter spp. were then matched for age and gender with patients hospitalised for gastroenteritis of other or unknown aetiology. Patients who had undergone abdominal radiographic investigation or had received a surgery consultation were included as "acute abdomen" (AA) cases. Demographics, clinical characteristics and management were compared between AA and non-AA cases. RESULTS: One hundred and forty-one patients with cultures positive for Campylobacter spp. were included in the analysis. Nineteen patients were identified as AA cases. Fewer of these had diarrhoea (14/19, 74% vs 117/121, 97%; p = 0.02) and more reported a lower sense of general wellbeing (8/18, 44% vs 8/108, 7%; p <0.001). Localised pain (9/18, 50% vs 20/115, 17%; p = 0.002) and abdominal tenderness (2/18, 11% vs 0/111; p = 0.02) were also more common among AA cases. Forty-four patients with Campylobacter spp. infections were hospitalised and matched with 44 patients with gastroenteritis of other or unknown aetiology. Campylobacter spp. infection (risk ratio 3.6, 95% CI 1.3-9.7; p = 0.01) was positively correlated with being seen by a surgeon and/or a prescription for radiological examination. CONCLUSIONS: We identified a subset of patients with Campylobacter spp. gastroenteritis who present as an acute abdominal emergency. The presentation of these patients was characterised mainly by the nature of the associated abdominal pain.
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Infecciones por Campylobacter , Campylobacter , Enteritis , Gastroenteritis , Adolescente , Infecciones por Campylobacter/complicaciones , Niño , Diarrea , Enteritis/complicaciones , Gastroenteritis/complicaciones , Humanos , Estudios RetrospectivosRESUMEN
Objectives: Clostridioides difficile infection (CDI) is the leading cause of healthcare-associated diarrhea, often complicated by severe infection and recurrence with increased morbidity and mortality. Data from large cohorts in Switzerland are scarce. We aimed to describe diagnostic assays, treatment, outcomes, and risk factors for CDI in a large cohort of patients in Switzerland. Methods: We conducted a retrospective cohort study of CDI episodes diagnosed in patients from two tertiary care hospitals in Switzerland. During a 3-month follow-up, we used a composite outcome combining clinical cure at day 10, recurrence at week 8, or death, to evaluate a patient's response. Unfavorable outcomes consisted in the occurrence of any of these events. Results: From January 2014 to December 2018, we included 826 hospitalized patients with documented CDI. Overall, 299 patients (36.2%) had a severe infection. Metronidazole was used in 566 patients (83.7%), compared to 82 patients (12.1%) treated with vancomycin and 28 patients (4.1%) treated with fidaxomicin. Overall mortality at week 8 was at 15.3% (112/733). Eighty-six patients (12.7%) presented with clinical failure at day 10, and 78 (14.9%) presented with recurrence within 8 weeks; 269 (39.8%) met the composite outcome of death, clinical failure, or recurrence. The Charlson Comorbidity Index score (p < 0.001), leukocytes > 15 G/L (p = 0.008), and the use of metronidazole (p = 0.012) or vancomycin (p = 0.049) were factors associated with the composite outcome. Conclusions: Our study provides valuable insights on CDI treatment and outcomes in Switzerland, highlights the heterogeneity in practices among centers, and underlines the need for the active monitoring of clinical practices and their impact on clinical outcomes through large multicentric cohorts.