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1.
J Fungi (Basel) ; 9(2)2023 Jan 24.
Article in English | MEDLINE | ID: mdl-36836274

ABSTRACT

Several studies have evaluated the serum galactomannan (GM) antigen assay in pediatric patients, and there is convincing evidence for its usefulness as a diagnostic tool for invasive Aspergillus infections in patients with acute leukemias or post allogeneic hematopoietic cell transplantation (HCT). Less is known about the utility of the assay in monitoring responses to treatment in patients with established invasive aspergillosis (IA). Here, we present the long-term kinetics of serum galactomannan in two severely immunocompromised adolescents with invasive pulmonary aspergillosis (IPA) who were cured after complicated clinical courses. We also review the utility of the GM antigen assay in serum as a prognostic tool around the time of diagnosis of IA and as a biomarker to monitor disease activity in patients with established IA and assess responses to systemic antifungal therapy.

2.
Microbiol Spectr ; 10(3): e0014822, 2022 06 29.
Article in English | MEDLINE | ID: mdl-35435751

ABSTRACT

The ability of extensively drug-resistant (XDR) Klebsiella pneumoniae to rapidly acquire resistance to novel antibiotics is a global concern. Moreover, Klebsiella clonal lineages that successfully combine resistance and hypervirulence have increasingly occurred during the last years. However, the underlying mechanisms of counteracting fitness costs that accompany antibiotic resistance acquisition remain largely unexplored. Here, we investigated whether and how an XDR sequence type (ST)307 K. pneumoniae strain developed resistance against the novel drug combination ceftazidime-avibactam (CAZ-AVI) using experimental evolution. In addition, we performed in vitro and in vivo assays, molecular modeling, and bioinformatics to identify resistance-conferring processes and explore the resulting decrease in fitness and virulence. The subsequent amelioration of the initial costs was also addressed. We demonstrate that distinct mutations of the major nonselective porin OmpK36 caused CAZ-AVI resistance that persists even upon following a second experimental evolution without antibiotic selection pressure and that the Klebsiella strain compensates the resulting fitness and virulence costs. Furthermore, the genomic and transcriptomic analyses suggest the envelope stress response regulator rpoE and associated RpoE-regulated genes as drivers of this compensation. This study verifies the crucial role of OmpK36 in CAZ-AVI resistance and shows the rapid adaptation of a bacterial pathogen to compensate fitness- and virulence-associated resistance costs, which possibly contributes to the emergence of successful clonal lineages. IMPORTANCE Extensively drug-resistant Klebsiella pneumoniae causing major outbreaks and severe infections has become a significant challenge for health care systems worldwide. Rapid resistance development against last-resort therapeutics like ceftazidime-avibactam is a significant driver for the accelerated emergence of such pathogens. Therefore, it is crucial to understand what exactly mediates rapid resistance acquisition and how bacterial pathogens counteract accompanying fitness and virulence costs. By combining bioinformatics with in vitro and in vivo phenotypic approaches, this study revealed the critical role of mutations in a particular porin channel in ceftazidime-avibactam resistance development and a major metabolic regulator for ameliorating fitness and virulence costs. These results highlight underlying mechanisms and contribute to the understanding of factors important for the emergence of successful bacterial pathogens.


Subject(s)
Klebsiella Infections , Klebsiella pneumoniae , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Azabicyclo Compounds , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Ceftazidime , Drug Combinations , Humans , Klebsiella Infections/drug therapy , Klebsiella Infections/microbiology , Klebsiella pneumoniae/genetics , Klebsiella pneumoniae/metabolism , Microbial Sensitivity Tests , Porins , Virulence/genetics , beta-Lactamases/genetics
3.
Microorganisms ; 10(1)2022 Jan 08.
Article in English | MEDLINE | ID: mdl-35056579

ABSTRACT

Vancomycin-resistant enterococci (VRE) pose a public health challenge worldwide. While VRE bloodstream infections (VREBI) increase in Germany and Europe, population-based molecular data are scarce. We aimed to analyze the molecular epidemiology, demographic aspects, and geographical distribution of VREBI in the German Federal State of North-Rhine-Westphalia (NRW), located in the German-Dutch-Belgian border area, representing over 20% of Germany's population. VREBI isolates were collected from hospitals across NRW between 2016 and 2019. Demographic data were gathered and anonymized upon sample collection. Multilocus sequence typing (MLST) and identification of glycopeptide resistance were carried out. Epidemiological analysis and geographical mapping were performed. Single VREBI isolates from 755 patients were analyzed. In total, 38.9% were female, and 80.0% were aged ≥ 60 years. The VREBI incidence per 100,000 inhabitants nearly tripled, from 0.52 (2016) to 1.48 (2019), particularly in male patients aged ≥ 50 years. The proportion of vanB reached 83% (n = 202/243) in 2018, overtaking vanA as the predominant glycopeptide resistance determinant, detected in close relation with ST117 isolates. The proportion of MLST sequence type (ST) 117 peaked in 2018, at 78.2% (n = 190/243). The major role of these emerging strains in invasive infections in central Europe requires novel strategies for their diagnosis, treatment, and prevention.

4.
Pharmaceuticals (Basel) ; 14(11)2021 Oct 21.
Article in English | MEDLINE | ID: mdl-34832848

ABSTRACT

Vancomycin is frequently used for the treatment of C. difficile infections (CDI). There are concerns that this might increase the risk of selecting vancomycin resistant enterococci (VRE). Here, we evaluated whether there is an increased risk of VRE acquisition following vancomycin for CDI specific treatment. Patients with CDI, metronidazole, or oral vancomycin treatment and without preexisting VRE were monitored for VRE acquisition. VRE isolates from patients with acquired and preexisting colonization were collected and subjected to whole genome sequencing. In total, 281 patients (median age 56 years, 54% of the male sex) presented with toxin positive C. difficile. Of them, 170 patients met the inclusion criteria, comprising 37 patients treated with metronidazole and 133 treated with oral vancomycin. In total, 14 patients meeting the inclusion criteria acquired VRE (vancomycin: n = 11; metronidazole: n = 3). Statistical analysis revealed no significant differences between both VRE acquisition rates. Genetic comparison of detected VRE isolates resulted in eight clusters of closely related genotypes comprising acquired and preexisting strains. Our results suggest that vancomycin and metronidazole likewise increase the risk of VRE acquisition. Genetic comparison indicates that VRE acquisition is a result of both antibiotic selection and pathogen transmission.

5.
Microbiol Spectr ; 9(2): e0035621, 2021 10 31.
Article in English | MEDLINE | ID: mdl-34523992

ABSTRACT

Vancomycin-resistant enterococci (VRE) are nosocomial pathogens with increasing prevalence worldwide. Extensive hygiene measures have been established to prevent infection transmission in hospitals. Here, we developed a predictive score system (the predictive vancomycin-resistant enterococci [PREVENT] score) to identify the clearance or persistence in patients with a history of VRE carrier status at readmission. Over a cumulative 3-year period, patients with a positive VRE carrier status were included. The study population was recruited in two successive time periods and separated into training data for predictive score development and validation data for evaluation of the predictive power. The risk factors for persistent VRE colonization were analyzed in a univariable analysis before development of a logistic regression model based on the potential risk factors. The score points were determined proportionally to the beta coefficients of the logistic regression model. The data from 448 (79%) patients were used as the training data, and those from 119 (21%) as the validation data. Multivariable analysis revealed the following variables as independent risk factors: age of ≥60 years, hemato-oncological disease, cumulative antibiotic treatment for >4 weeks, and a VRE infection. The resulting logistic regression model exhibited an acceptable area under the curve (AUC) of 0.81 (95% confidence interval [CI], 0.72 to 0.91). The predictive score system had a sensitivity of 82% (95% CI, 65 to 93%) and a specificity of 77% (95% CI, 66 to 85%). The developed predictive score system is a useful tool to assess the VRE carrier status of patients with a history of VRE colonization. On the basis of this risk assessment, more focused and cost-effective infection control measures can be implemented. IMPORTANCE Given the increasing relevance of VRE as nosocomial pathogens worldwide, infection prevention and control measures, including patient isolation and contact precautions, are indispensable to avoid their spread in the hospital setting. In this study, we developed and validated the PREVENT score, a tool for rapid risk assessment of VRE persistence in patients with a history of previous VRE colonization. The score is designed to be easily performed, employing clinical information available in a regular admission setting and immediately providing information to inform the decision of whether to adopt patient isolation and contact precautions during the hospital stay. After validation, the score was shown to accurately identify patients with persistent VRE colonization upon admission, representing a suitable option as (i) a complementary method yielding preliminary results significantly more quickly than culture-based VRE detection techniques and (ii) an alternative strategy for VRE detection in settings in which microbiological VRE screening is not routinely performed due to limited resources.


Subject(s)
Cross Infection/prevention & control , Gram-Positive Bacterial Infections/epidemiology , Infection Control/methods , Primary Prevention/methods , Vancomycin-Resistant Enterococci/isolation & purification , Cross Infection/microbiology , Female , Gram-Positive Bacterial Infections/diagnosis , Gram-Positive Bacterial Infections/microbiology , Humans , Logistic Models , Male , Middle Aged , Persistent Infection/diagnosis , Persistent Infection/epidemiology , Persistent Infection/microbiology , Risk Assessment/methods , Risk Factors
6.
Biol Sex Differ ; 12(1): 36, 2021 05 17.
Article in English | MEDLINE | ID: mdl-34001270

ABSTRACT

BACKGROUND: Vancomycin-resistant enterococci (VRE) have emerged in the healthcare setting worldwide. Infections with these pathogens, i.e., bloodstream infections (BSI), are accompanied with an impaired patient outcome. Diverse factors comprising patient characteristics, therapeutic strategies, and infection control measures are positively or negatively associated with VRE BSI occurrence. However, whether sex-specific differences influence the frequency of VRE BSI is yet unknown. The aim of this systematic review was to comprehensively summarize and analyze sex prevalence in VRE BSI patients. MAIN TEXT: A systematic search for relevant articles was conducted in PubMed and Web of Science. After screening for eligibility, data extraction from included articles and risk of bias assessment were processed. The prevalence of male/female sex in VRE BSI patients and 95% CI were calculated for each study and summarized as pooled estimated effect. In total, nine articles met the inclusion criteria. Risk of bias assessment resulted in low (six studies) to moderate bias (three studies). The pooled prevalence of male patients suffering from VRE BSI was 59% resulting in a 1.4 male/female prevalence ratio. CONCLUSIONS: Current literature suggests sex differences with male preference (59%) in the distribution of VRE BSI cases. Further primary studies should address the question of male-specific factors favoring the enhanced frequency of VRE BSI.


Subject(s)
Bacteremia , Gram-Positive Bacterial Infections , Vancomycin-Resistant Enterococci , Anti-Bacterial Agents , Bacteremia/drug therapy , Bacteremia/epidemiology , Female , Gram-Positive Bacterial Infections/drug therapy , Gram-Positive Bacterial Infections/epidemiology , Humans , Male , Sex Characteristics
7.
Antimicrob Resist Infect Control ; 9(1): 180, 2020 11 07.
Article in English | MEDLINE | ID: mdl-33160394

ABSTRACT

BACKGROUND: Transplant recipients are at high risk for infections. However, donor-recipient transmission of multidrug-resistant organisms (MDROs) remains mostly unaddressed in the protocols of pre-transplant infection and colonization screening. Vancomycin-resistant enterococci (VRE) are MDROs that colonize the gastrointestinal tract and are associated with a significant burden of disease. Besides the high mortality of invasive VRE infections, chronic colonization leads to costly isolation measures in the hospital setting. Whereas most post-transplantation VRE infections are endogenous and thus preceded by colonization of the recipient, conclusive evidence of VRE transmission via allograft in the context of intestinal transplantation is lacking. CASE PRESENTATION: We describe a donor-derived VRE infection after intestinal transplantation including small bowel and right hemicolon. The recipient, a 54-year old male with history of mesenteric ischemia and small bowel perforation due to generalized atherosclerosis and chronic stenosis of the celiac trunk and the superior mesenteric artery, developed an intra-abdominal infection and bloodstream infection after transplantation. VRE isolates recovered from the patient as well as from the allograft prior to transplantation were analyzed via whole genome sequencing. Isolates showed to be genetically identical, thus confirming the transmission from donor to recipient. CONCLUSIONS: This case underlines the relevance of donor-recipient VRE transmission and invasive infection in the context of intestinal transplantation, highlighting the need for preoperative MDRO screening that facilitates the prompt and effective treatment of possible infections as well as the timely establishment of contact precautions to prevent further spread.


Subject(s)
Bacteremia/etiology , Gram-Positive Bacterial Infections/transmission , Intestines/transplantation , Postoperative Complications/etiology , Tissue Donors , Vancomycin-Resistant Enterococci , Drug Resistance, Multiple, Bacterial , Humans , Male , Middle Aged
8.
Antimicrob Resist Infect Control ; 9(1): 154, 2020 09 22.
Article in English | MEDLINE | ID: mdl-32962759

ABSTRACT

BACKGROUND: Currently, hospitals have been forced to divert substantial resources to cope with the ongoing coronavirus disease 2019 (COVID-19) pandemic. It is unclear if this situation will affect long-standing infection prevention practices and impact on healthcare associated infections. Here, we report a nosocomial cluster of vancomycin-resistant enterococci (VRE) that occurred on a COVID-19 dedicated intensive care unit (ICU) despite intensified contact precautions during the current pandemic. Whole genome sequence-based typing (WGS) was used to investigate genetic relatedness of VRE isolates collected from COVID-19 and non-COVID-19 patients during the outbreak and to compare them to environmental VRE samples. METHODS: Five VRE isolated from patients (three clinical and two screening samples) as well as 11 VRE and six vancomycin susceptible Enterococcus faecium (E. faecium) samples from environmental sites underwent WGS during the outbreak investigation. Isolate relatedness was determined using core genome multilocus sequence typing (cgMLST). RESULTS: WGS revealed two genotypic distinct VRE clusters with genetically closely related patient and environmental isolates. The cluster was terminated by enhanced infection control bundle strategies. CONCLUSIONS: Our results illustrate the importance of continued adherence to infection prevention and control measures during the COVID-19 pandemic to prevent VRE transmission and healthcare associated infections.


Subject(s)
Coinfection/drug therapy , Cross Infection/epidemiology , Gram-Positive Bacterial Infections/drug therapy , Gram-Positive Bacterial Infections/epidemiology , Vancomycin-Resistant Enterococci/drug effects , Betacoronavirus , COVID-19 , Coinfection/microbiology , Coronavirus Infections/pathology , Cross Infection/drug therapy , Cross Infection/microbiology , Disease Outbreaks , Genome, Bacterial/genetics , Gram-Positive Bacterial Infections/microbiology , Humans , Infection Control , Intensive Care Units , Multilocus Sequence Typing , Pandemics , Pneumonia, Viral/pathology , Primary Prevention , SARS-CoV-2 , Vancomycin-Resistant Enterococci/genetics , Vancomycin-Resistant Enterococci/isolation & purification , Whole Genome Sequencing
9.
Microorganisms ; 8(9)2020 Sep 08.
Article in English | MEDLINE | ID: mdl-32911751

ABSTRACT

During the current severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, healthcare systems worldwide have to prevent nosocomial SARS-CoV-2 transmission while maintaining duty of care. In our study, we characterize the transmission dynamic of SARS-CoV-2 in inpatients and healthcare workers (HCWs) at the University Hospital Münster (UHM) in northwest Germany. We identified 27 cases of healthcare-associated SARS-CoV-2 infections (4 inpatients and 23 HCWs) who had contact with patients and/or HCWs without the use of adequate PPE. The contacts of these index cases were followed up for SARS-CoV-2 infection after unprotected exposure and a quantitative measure of probability of becoming infected, the attack rate, was calculated. In addition, transmission was evaluated in the context of infection control measures established during the pandemic and we compared the epidemiological data of all index cases, including symptoms and Ct values of virology test results. The overall attack rate in the hospital setting was 1.3% (inpatients 0.9%, HCWs 1.6%). However, during an outbreak, the attack rate was 25.5% (inpatients 20.0%, HCWs 29.6%). For both scenarios, HCWs had a higher attack rate illustrating their role in healthcare-associated SARS-CoV-2 transmission. Taken together, our experiences demonstrate how infection control measures can minimize the transmission of SARS-CoV-2 in the healthcare setting.

10.
Travel Med Infect Dis ; 37: 101696, 2020.
Article in English | MEDLINE | ID: mdl-32344112

ABSTRACT

BACKGROUND: Traveller's diarrhea (TD) is the leading cause of morbidity among travelers. Recently, molecular methods have been developed to detect the causative pathogens of TD. The aim of this study was to test a multiplex-PCR approach in patients with TD and asymptomatic controls. METHODS: In total, 91 travelers (61 TD cases, 30 asymptomatic controls) prospectively collected stool samples during travel and documented gastrointestinal symptoms. Samples were analyzed using the BioFire® FilmArray® Gastrointestinal Panel, which covers 13 enteric bacteria (e.g. diarrheagenic Escherichia coli), 4 protozoan parasites and 5 viruses. RESULTS: TD cases had more likely a positive test (46/61, 75%) compared to controls (17/30, 57%, p = 0.07). The most common enteric pathogens were enteroaggregative E. coli (36/91, 40%), followed by enterotoxigenic E. coli (31/91, 34%) and enteropathogenic E. coli (EPEC, 29/91, 32%). The only pathogens significantly associated with TD were EPEC (p = 0.01) and ETEC (p = 0.047). Protozoan parasites were not detected. CONCLUSION: Our results challenge the widespread use of multiplex-approaches covering numerous enteric pathogens in the diagnostic workup of TD because only few (i.e. diarrheagenic E. coli) were related to symptoms of TD.


Subject(s)
Diarrhea/etiology , Enteropathogenic Escherichia coli , Escherichia coli Infections , Travel-Related Illness , Case-Control Studies , Diarrhea/epidemiology , Feces , Humans , Multiplex Polymerase Chain Reaction
12.
Sci Rep ; 10(1): 4988, 2020 03 19.
Article in English | MEDLINE | ID: mdl-32193431

ABSTRACT

Carbapenemase-producing bacteria are a growing issue worldwide. Most phenotypic detection methods are culture-based, requiring long incubation times. We present a phenotypic screening panel for detection of carbapenem non-susceptibility and differentiation of carbapenemase classes and AmpC, the MALDI-TOF MS-based direct-on-target microdroplet growth assay (DOT-MGA). It was validated on 7 reference strains and 20 challenge Enterobacterales isolates. Broth microdilution (BMD) and combination disk test (CDT) were also performed, as well as PCR as reference method. The panel based on the synergy between meropenem and carbapenemase inhibitors, determined by incubating these substances with bacterial suspension on a MALDI-TOF MS target and subsequently assessing bacterial growth on the target's spots by MS. After 4 hours of incubation, DOT-MGA correctly identified KPC, MBL and OXA (100% agreement with PCR). Detection of AmpC coincided with BMD and CDT but agreement with PCR was low, not ruling out false negative PCR results. DOT-MGA delivered more accurate results than BMD and CDT in a significantly shorter time, allowing for detection of carbapenem non-susceptibility, MIC determination and carbapenemase differentiation in one step.


Subject(s)
Bacterial Proteins/analysis , Enterobacteriaceae/enzymology , Microbial Sensitivity Tests , beta-Lactamases/analysis , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/antagonists & inhibitors , Drug Resistance, Bacterial , Enterobacteriaceae/drug effects , Enzyme Inhibitors/pharmacology , Meropenem/pharmacology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods
13.
Microorganisms ; 8(2)2020 Jan 31.
Article in English | MEDLINE | ID: mdl-32024001

ABSTRACT

Vancomycin-resistant enterococci (VRE) are relevant nosocomial pathogens with an increasing incidence in the last decades. Their transmission is optimal in the hospital setting, as it offers two potential, large reservoirs that are closely related: susceptible patients and their environment. Here we investigate the role of the hospital environment in the nosocomial transmission of VRE by establishing concrete links between contaminated surfaces and colonized/infected patients in outbreak and non-outbreak settings. Environmental and patient VRE isolates were collected between 2013 and 2019 and analyzed by whole-genome sequencing (WGS), subsequent multilocus sequence typing (MLST), and core genome (cg) MLST. Pairs of isolates differing in <3 alleles were rated as closely related, making a transmission likely. Fifty-three environmental VRE isolates were analyzed. MLST sequence types (ST) ST203 (50.0%), ST192 (21.3%), ST117 (17.3%), ST721 (8.8%), ST80 (2%), and ST1489 (0.7%) were detected, carrying the resistance determinants vanA (72.7%), vanB (24%), or both (3.3%). Of the 53 environmental isolates, 51 were found to form five clusters with genetically related patient isolates (n = 97 isolates). WGS confirms the role of the environment in the transmission dynamics of VRE in both the outbreak and non-outbreak settings, highlighting the importance of prevention and control of VRE spread.

14.
BMC Infect Dis ; 19(1): 836, 2019 Oct 11.
Article in English | MEDLINE | ID: mdl-31601195

ABSTRACT

BACKGROUND: Moraxella nonliquefaciens is a usually non-pathogenic biofilm-producing Gram-negative coccobacillus which may colonize the upper respiratory tract, rarely causing invasive disease. Although very rare, bloodstream infections caused by this organism have been described, showing often a fatal outcome. Here, we report the case of a pediatric cancer patient with bloodstream infection and sepsis due to M. nonliquefaciens showing full recovery after appropriate antibiotic treatment. CASE PRESENTATION: A three-year-old boy with stage IV neuroblastoma was admitted for high-dose chemotherapy with autologous stem cell rescue after standard neuroblastoma treatment. Despite receiving antimicrobial prophylaxis with trimethoprim/sulfamethoxazole, acyclovir and amphothericin B, the patient presented with fever of up to 39.5 °C and neutropenia. Besides a chemotherapy-related mucositis and an indwelling Broviac catheter (removed), no infection focus was identified on physical examination. Moraxella nonliquafaciens was identified in blood cultures. After antibiotic treatment and neutrophil recovery, the patient was fit for discharge. CONCLUSIONS: The case described highlights the importance of an otherwise non-pathogenic microorganism, especially in immunosupressed cancer patients. It should be kept in mind that, although very infrequently, Moraxella nonliquefaciens may cause bloodstream infections that can be successfully treated with prompt focus identification and antibiotic therapy.


Subject(s)
Bacteremia/diagnosis , Moraxella/isolation & purification , Neoplasms/pathology , Sepsis/diagnosis , Anti-Bacterial Agents/therapeutic use , Antineoplastic Agents/therapeutic use , Bacteremia/complications , Bacteremia/drug therapy , Bacteremia/microbiology , Child, Preschool , Humans , Male , Neoplasm Staging , Neoplasms/complications , Neoplasms/drug therapy , Neutropenia/diagnosis , Neutropenia/etiology , Sepsis/etiology , Sepsis/microbiology
15.
Microorganisms ; 7(10)2019 Sep 26.
Article in English | MEDLINE | ID: mdl-31561632

ABSTRACT

Vancomycin-resistant enterococci (VRE) are important nosocomial pathogens that require effective infection control measures, representing a challenge for healthcare systems. This study aimed at identifying risk factors associated with prolonged VRE carriage and determining the rate of clearance that allows the discontinuation of contact precautions. During a 2-year study, screening was performed in patients with a history of VRE or at risk of becoming colonized. After bacterial identification and antibiotic susceptibility testing, glycopeptide resistance was confirmed by PCR. Isolates were compared via whole genome sequence-based typing. Risk factors were recorded, and follow-up screening was performed upon readmission, defining patients as long-term carriers if still colonized ≥10 weeks after first detection. Of 1059 patients positive for VRE, carriage status was assessed upon readmission in 463 patients. VRE was cleared in 56.4% of the cases. Risk factors associated with long-term persistence were hospital stays (frequency, length), hemato-oncological disease, systemic treatment with steroids, and use of antibiotics. No specific genotypic clustering was observed in patients with VRE clearance or persistence. VRE clearance is possibly underestimated. The identification of risk factors favoring long-term carriage may contribute to a targeted implementation of infection control measures upon readmission of patients with history of VRE.

16.
Front Microbiol ; 10: 13, 2019.
Article in English | MEDLINE | ID: mdl-30733710

ABSTRACT

Introduction: Antibiotic resistant bacteria are a growing concern worldwide. Extended-spectrum ß-lactamases (ESBL) represent the most common resistance mechanism of Gram-negative bacteria against ß-lactams, underlining the need for adequate diagnostic methods that provide reliable information in the shortest time possible. AmpC, a less prevalent but increasingly relevant class of ß-lactamases, pose an additional challenge as their detection is complex. Here, we present an ESBL and AmpC screening panel employing the MALDI-TOF MS-based direct-on-target microdroplet growth assay (DOT-MGA). Materials and Methods: Four reference strains recommended by the European Committee on Antimicrobial Susceptibility Testing (EUCAST) were used to develop the panel, which was further validated on 50 clinical Enterobacterales isolates resistant to third generation cephalosporins. The panel relies on the synergistic effect between ESBL and/or AmpC ß-lactamase inhibitors and cephalosporins, which indicates ß-lactamase production. Microdroplets containing the tested microorganism, cephalosporins in different concentrations and inhibitors were pipetted onto an MBT Biotarget and incubated for 3 or 4 h at 35 ± 1°C. Afterward, the liquid medium was removed and the material adhered to the spots was analyzed by MALDI-TOF MS. Synergy was detected by determining and comparing the minimum inhibitory concentrations of the tested cephalosporins with and without ß-lactamase inhibitors. Data were interpreted following a diagnostic algorithm proposed by EUCAST in order to establish a final diagnosis. In comparison, PCR, broth microdilution (BMD) and combination disk tests (CDT) were performed. Results: Compared to the PCR results, the following positive and negative percent agreement values (PPA/NPA) were obtained for each resistance mechanism: ESBL, 94.44/100%; AmpC, 94.44/93.75% and ESBL+AmpC, 100/100%. These results, obtained after 4 h of incubation, were comparable with those of BMD and showed a higher accuracy than CDT. Discussion: We propose a novel phenotypic method for detection of ESBL and AmpC ß-lactamases in Enterobacterales that provides reliable results in a short time, representing a promising alternative to the diagnostic techniques currently available. This easy-to-perform approach has potential for being implemented in routine laboratories, contributing to the further diversification of mass spectrometry technology into other fields such as antibiotic resistance testing.

17.
Rev. med. Risaralda ; 18(1): 88-93, jun. 2012.
Article in Spanish | LILACS | ID: lil-649077

ABSTRACT

Introducción: La escabiosis o sarna humana es una dermatosis causada a partir de la infección cutánea por Sarcoptes scabiei. Una variante severa y rara de ella es la escabiosis noruega o costrosa, que ha repuntado en los últimos años por el incremento en patologías que causan inmunosupresión. Clínicamente se presenta con hiperqueratosis y placas costrosas predominantemente en tórax, cabeza, palmas y plantas. Su diagnóstico es sencillo, mediante la observación microscópica del artrópodo. Se ha descrito su tratamiento con permetrina, benzoato de bencilo e ivermectina. Caso: Paciente masculino de 38 años de edad, con antecedente de sarna noruega tratada hace 2 años y hábito alcohólico acentuado. Inicia enfermedad actual hace 2 meses, caracterizada por pápulas pruriginosas generalizadas que aumentaron progresivamente en cantidad, formando placas costrosas. Examen físico: xerosis cutis, placas hiperqueratósicas de variadas formas y tamaños, algunas coalescentes, fácilmente desprendibles, sobre base eritematosa, brillante, rezumante, localizadas en cuello, tronco y extremidades. Examen microscópico: huevos y adultos de Sarcoptes scabiei. Tratamiento: cefazolina, maleato de clorfeniramina, cetirizina, ivermectina. Discusión: La sarna noruega recurrente en un paciente adulto sin patología de base conocida, debe conllevar a la búsqueda de causas que comprometan la respuesta inmune del individuo. Más allá de las patologías médicas y medicamentos con efectos inmunosupresores conocidos, debe también indagarse el consumo habitual sustancias como alcohol, agente con demostradas propiedades supresoras sobre la respuesta inmune innata y adquirida. En este caso, el tratamiento dermatológico debe ser complementado con el manejo adecuado del hábito alcohólico subyacente a la patología del paciente.


Background: Scabies is a dermatosis caused by infection due to Sarcoptes scabiei. An uncommon and severe form of this condition, Norwegian or crusted scabies, has shown an increasing incidence in recent years, due to the growing number of pathologies causing immunosupression. Clinical presentation includes hyperkeratosis and crusted plaques mainly on the skin of the thorax, head, palms and soles. It is easily diagnosed by direct observation of the arhtropode. Permethrin, benzyl benzoate and ivermectin have been described as effective therapeutic resources. Case: A 38-year-old male patient, with previous history of Norwegian scabies treated 2 years ago, and heavy consumption of alcohol is described. Presented illness begins 2 months ago, with the appearance of generalized pruriginous papules which progressively grew into crusted plaques. Physical examination: xerosis cutis, hyperkeratosic plaques in different shapes and sizes, some of them converging, easily removable, on an erytematous, bright, humid base, located on the skin of the neck, torso and limbs. Microscopic examination: eggs and adult forms of Sarcoptes scabiei. Treatment: cefazolin, clorpheniramine, cetirizin, ivermectin. Discussion: Recurrent Norwegian scabies in an adult patient without history of a previous disease, must be a reason to find other causes that jeopardize the individual’s immune response. Besides medical conditions and drugs known to have immunosuppressing effects, habits such as alcohol consumption must be interrogated, for it has been demonstrated that this substance has suppressing effects on innate and acquired immunity. The dermatologic treatment must be complimented with an accurate management of the alcohol abuse which underlies the disease.


Subject(s)
Humans , Alcoholism , Mite Infestations , Sarcoptes scabiei , Dermatology , Venezuela
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