RESUMEN
OBJECTIVES: To gain data on the current molecular epidemiology and resistance of MRSA in the Czech Republic. METHODS: Between September 2017 and January 2018, a total of 441 single-patient MRSA isolates were collected from 11 Czech hospitals and analysed by spa typing, SCCmec typing, antibiotic susceptibility testing, detection of the PVL toxin and the arcA gene. RESULTS: Of all MRSA isolates, 81.41% (n = 359) belonged to the CC5-MRSA clone represented by the spa types t003 (n = 136), t586 (n = 92), t014 (n = 81), t002 (n = 20) and other spa types (n = 30); a majority of the CC5 isolates (n = 348, 96.94%) carried SCCmec type II. The occurrence of CC5-MRSA was more likely in older inpatients and associated with a healthcare origin (P < 0.001). The CC5-MRSA isolates were resistant to more antimicrobial drugs compared with the other MRSAs (P < 0.001). Interestingly, t586 was detected in blood samples more often than the other spa types and, contrary to other spa types belonging to CC5-MRSA, t586 was not associated with patients of advanced age. Other frequently found lineages were CC8 (n = 17), CC398 (n = 11) and CC59 (n = 10). The presence of the PVL was detected in 8.62% (n = 38) of the MRSA isolates. CONCLUSIONS: The healthcare-associated CC5-MRSA-II lineage (t003, t586, t014) was found to be predominant in the Czech Republic. t586 is a newly emerging spa type in the Czech Republic, yet reported rarely in other countries. Our observations stress the need for MRSA surveillance in the Czech Republic in order to monitor changes in MRSA epidemiology.
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Staphylococcus aureus Resistente a Meticilina , Infecciones Estafilocócicas , Anciano , Antibacterianos/farmacología , República Checa/epidemiología , Genotipo , Humanos , Staphylococcus aureus Resistente a Meticilina/genética , Pruebas de Sensibilidad Microbiana , Epidemiología Molecular , Infecciones Estafilocócicas/epidemiologíaRESUMEN
BACKGROUND: Colistin is a last-resort antibiotic used for the treatment of infections caused by multidrug-resistant Gram-negative pathogens. The emergence of plasmid-borne colistin resistance mediated by the mcr genes poses a risk of its spread and its occurrence should be monitored. The aim of this study was to discuss possible detection methods and their reliability in screening for this type of resistance. MATERIAL AND METHODS: The reliability of the disk diffusion method, Rapid Polymyxin NP test and two types of gradient tests for the screening of mcr-mediated colistin resistance was evaluated on 16 human and two reference isolates of Escherichia coli (colistin-susceptible and colistin-resistant with chromosomally-mediated resistance or harboring the mcr-1 gene). Broth microdilution was the reference method used for the determination of colistin resistance. RESULTS: Targeted screening for colistin-resistant strains is best performed with a selective agar medium supplemented with colistin. In cultured isolates, suspected colistin resistance should always be confirmed by reliable methods. There was 100 percent agreement between both the gradient methods and the Rapid Polymyxin NP test and the reference broth microdilution method. When using the disk diffusion method with 10µg and 50µg disks, only 11 % and 33 % of results were correct, respectively. Therefore, disk diffu-sion is inappropriate for colistin resistance screening. CONCLUSION: Prospective prevalence studies of intestinal carriage of colistin-resistant Enterobacterales among Czech hospitalized patients and travelers do not yet indicate the spread of strains with mcr-mediated plasmid-borne colistin resistance. However, the high prevalence of strains carrying the mcr genes in raw meat products poses a risk of exposure for their consumers. The detection of mcr-harbouring colistin-resistant strains cannot be expected during routine microbiological testing, without using reliable but expensive methods. A suitable alternative method for active monitoring of colistin resistance is the use of selective agar media with colistin followed by verification of the resistance in the obtained isolates.
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Colistina , Farmacorresistencia Bacteriana , Antibacterianos/farmacología , Colistina/farmacología , Farmacorresistencia Bacteriana/genética , Humanos , Pruebas de Sensibilidad Microbiana , Plásmidos/genética , Estudios Prospectivos , Reproducibilidad de los ResultadosRESUMEN
Clostridioides (Clostridium) difficile infection (CDI) is the most common causative pathogen of health care-associated gastrointestinal infections; however, due to the overlap of clinical symptoms with those of other causes of acute gastroenteritis, the selection of the most appropriate laboratory test is difficult. From April to October 2018, 640 stool samples requested for CDI testing were examined using the mariPOC CDI and Gastro test (ArcDia), which allows the detection of C. difficile glutamate dehydrogenase (GDH) and toxin A/B, norovirus genogroups GI and GII.4, rotavirus, adenovirus, and Campylobacter spp. In parallel, the C. Diff Quik Chek Complete test (Alere) was used as a routine diagnostic assay, and C. difficile toxigenic culture was used as a reference method. The sensitivity of the mariPOC CDI and Gastro test was comparable to that of C. Diff Quik Chek Complete for the detection of GDH (96.40% [95% confidence interval {CI}, 91.81% to 98.82%] versus 95.68% [95% CI, 90.84 to 98.40%]; P = 1.00) and was higher for the detection of toxin A/B (66.67% [95% CI, 57.36 to 75.11%] versus 55.56% [95% CI, 46.08 to 64.74%]; P = 0.00). The specificity of the mariPOC CDI and Gastro test was lower than that of C. Diff Quik Chek Complete for GDH detection (95.21% [95% CI, 92.96% to 96.91%] versus 97.60% [95% CI, 95.85% to 98.76%]; P = 0.04) and comparable to that of C. Diff Quik Chek Complete for toxin A/B detection (99.24 [95% CI, 98.05% to 99.79%] versus 99.81% [95% CI, 98.94% to 100.0%]; P = 0.37). In 29 cases (4.53%), other causative agents of diarrhea were detected (Campylobacter spp. [n = 17], rotavirus [n = 7], and norovirus genogroup GII.4 [n = 5]).
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Clostridioides difficile , Infecciones por Clostridium/diagnóstico , Infecciones por Clostridium/microbiología , Heces/microbiología , Inmunoensayo , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Proteínas Bacterianas/genética , Toxinas Bacterianas/genética , Niño , Preescolar , Clostridioides difficile/enzimología , Clostridioides difficile/genética , Clostridioides difficile/inmunología , Infecciones por Clostridium/inmunología , Pruebas Diagnósticas de Rutina , Manejo de la Enfermedad , Enterocolitis Seudomembranosa/diagnóstico , Enterocolitis Seudomembranosa/inmunología , Enterocolitis Seudomembranosa/microbiología , Enterotoxinas/genética , Femenino , Glutamato Deshidrogenasa , Humanos , Inmunoensayo/métodos , Inmunoensayo/normas , Lactante , Masculino , Persona de Mediana Edad , Sensibilidad y Especificidad , Adulto JovenRESUMEN
Clostridium difficile is a major nosocomial pathogen in humans with an increasing incidence in the community. The "one-health" approach of research is needed to investigate possible reservoirs of C. difficile and route of its transmission. The objective of this study is to investigate the occurrence of C. difficile in pigs in the Czech Republic with characterisation of the isolates to determine their genetic relatedness to C. difficile isolates from European and Asian pigs. A total of 198 pig faeces samples from 23 farms were investigated and of those 57 samples (55 piglets, 2 sows) from 11 farms were confirmed as C. difficile positive. The majority of C. difficile isolates belonged to the sequence type 11 and clade 5. The predominant ribotypes were 078 (nâ¯=â¯23), 078-variant (nâ¯=â¯5), 033 (nâ¯=â¯10) followed by RTs 150 (nâ¯=â¯7), 011 (nâ¯=â¯5), 045 (nâ¯=â¯4), 126, 014, 002 (nâ¯=â¯1, each). All isolates were susceptible to metronidazole, vancomycin and tetracycline. Isolates of RTs 150 and 078-variant were moxifloxacin resistant (MIC≥32â¯mg/L) and carried the amino acid substitution Thr82Ile in the GyrA. A multi-locus variable number tandem-repeats analysis (MLVA) revealed a clonal relatedness of isolates within individual farms and in C. difficile RT078 isolates between two Czech farms. Czech C. difficile RT078 isolates clustered with German C. difficile RT078 isolates and Czech C. difficile 078-variant isolates clustered with C. difficile RT078 isolates from Japan and Taiwan. This study found an emergence of C. difficile RT078 in Czech piglets that was related genetically to C. difficile RT078 isolates from Germany, Japan and Taiwan.
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Clostridioides difficile/clasificación , Clostridioides difficile/genética , Infecciones por Clostridium/transmisión , Infecciones por Clostridium/veterinaria , Sustitución de Aminoácidos/genética , Animales , Antibacterianos/farmacología , Clostridioides difficile/aislamiento & purificación , Infecciones por Clostridium/microbiología , República Checa , Girasa de ADN/genética , Alemania , Japón , Metronidazol/farmacología , Pruebas de Sensibilidad Microbiana , Moxifloxacino/farmacología , Tipificación de Secuencias Multilocus , Ribotipificación , Porcinos , Taiwán , Tetraciclina/farmacología , Vancomicina/farmacologíaRESUMEN
Bacteroides pyogenes can cause infections in humans. We describe a case of bloodstream infection caused by Bacteroides denticanum that probably originated from a dog bite. MALDI-TOF MS misidentified this new species as B. pyogenes. Subsequent analysis using the 16S rRNA sequencing approach identified the species as B. denticanum.
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Bacteriemia/microbiología , Infecciones por Bacteroides/microbiología , Bacteroides/aislamiento & purificación , Anciano , Animales , Bacteriemia/diagnóstico , Técnicas de Tipificación Bacteriana , Bacteroides/química , Bacteroides/clasificación , Bacteroides/genética , Infecciones por Bacteroides/diagnóstico , Perros , Femenino , Humanos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
Bacterial persistence in clinical microbiology is a phenomenon where the bacterial subpopulation of any bacterial strain, without having been exposed to an antibiotic, is already persistent to it. In clinical bacterial strains, persistence is not tested at all and the role of this phenomenon in the treatment of bacterial infections has not yet been evaluated. Therefore, the aim of the article is to highlight the significance of this probably global phenomenon in the treatment of bacterial infections with antibiotics. Also described are the mechanisms of its origin and some manner that could potentially reduce the frequency of these antibiotic-resistant bacterial cells in the bacterial population.
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Antibacterianos , Bacterias , Infecciones Bacterianas , Farmacorresistencia Bacteriana , Antibacterianos/farmacología , Bacterias/efectos de los fármacos , Infecciones Bacterianas/microbiología , HumanosRESUMEN
Bacillary angiomatosis (BA) is a disorder of neovascular proliferation involving skin and other organs of immunosuppressed patients caused by Bartonella species. BA has been recognized in both immunocompetent and immunodeficient patients, mostly in human immunodeficiency virus (HIV)-infected persons, much more rare in those with other immunodeficiencies, including organ transplantation. Diagnosis is based on serologic analysis, culture and molecular biology [detection of Bartonella species deoxyribonucleic acid (DNA) in tissue biopsy extracts by real-time polymerase chain reaction (PCR)]. All immunosuppressed patients with BA should be treated with antibiotics because of potentially life-threatening course of the disease. We report the first case of cutaneous bacillary angiomatosis due to Bartonella quintana in renal transplant recipient. This presentation demonstrates that BA should be considered a differential diagnosis in immunocompromised patients presenting with fever and cutaneous angioma-like lesions.
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Angiomatosis Bacilar/inmunología , Bartonella quintana , Trasplante de Riñón/efectos adversos , Adolescente , Adulto , Angiomatosis Bacilar/microbiología , Antibacterianos/uso terapéutico , Biopsia , Niño , ADN/química , Femenino , Humanos , Terapia de Inmunosupresión , Inmunosupresores/química , Fallo Renal Crónico/complicaciones , Fallo Renal Crónico/cirugía , Masculino , Persona de Mediana Edad , Complicaciones Posoperatorias , Reacción en Cadena en Tiempo Real de la Polimerasa , Adulto JovenRESUMEN
Antibiotic profiling of twenty Czech Clostridium difficile PCR-ribotype 176 isolates revealed a high level of resistance to erythromycin, ciprofloxacin and moxifloxacin (n = 20) and to rifampicin (n = 13). Accumulation of resistance mechanisms to multiple antibiotics highlight that PCR-ribotype 176 belong to problematic epidemic strains.
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Antibacterianos/farmacología , Clostridioides difficile/efectos de los fármacos , Farmacorresistencia Bacteriana Múltiple , Enterocolitis Seudomembranosa/microbiología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Secuencia de Bases , Clostridioides difficile/clasificación , Clostridioides difficile/genética , Clostridioides difficile/aislamiento & purificación , República Checa , Humanos , Datos de Secuencia Molecular , Mutación , RibotipificaciónRESUMEN
Following the COVID-19 infection, the sternum dislocation and wound dehiscence resulted in an infection complicating the recovery of an immunosuppressed patient after bilateral lung transplantation. Anaerobic culture (96 h) of milky cloudy wound secretion resulted in the growth of pinpoint haemolytic colonies identified as Metamycoplasma hominis (formerly Mycoplasma hominis). The search for the endogenous source of the infection found the bacterium exclusively in the patient's sputum, making a possible link to donor lung M. hominis colonization. Unfortunately, the donor samples were no longer available. The wound infection was successfully treated with 17 days of clindamycin despite the continuous PCR detection of M. hominis in the sputum after the end of the treatment.
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Trasplante de Pulmón , Infecciones por Mycoplasma , Mycoplasma hominis , Infección de la Herida Quirúrgica , Humanos , Trasplante de Pulmón/efectos adversos , Infección de la Herida Quirúrgica/microbiología , Infección de la Herida Quirúrgica/tratamiento farmacológico , Infección de la Herida Quirúrgica/diagnóstico , Mycoplasma hominis/genética , Mycoplasma hominis/aislamiento & purificación , Infecciones por Mycoplasma/microbiología , Infecciones por Mycoplasma/diagnóstico , Infecciones por Mycoplasma/tratamiento farmacológico , Masculino , COVID-19/diagnóstico , Antibacterianos/uso terapéutico , Esputo/microbiología , Persona de Mediana Edad , SARS-CoV-2/genética , SARS-CoV-2/aislamiento & purificación , Huésped Inmunocomprometido , Clindamicina/uso terapéuticoRESUMEN
Staphylococcus aureus is one of the most common bacterial pathogens, often asymptomatically colonizing healthy people, but capable of causing fatal disease. The ability to treat S. aureus infections is limited by the rapid spread of multidrug-resistant strains. This study aimed to determine the prevalence of S. aureus carriage among students from Okada, Edo State, Nigeria, to analyze the antibiotic resistance patterns and molecular characteristics of S. aureus isolates. One hundred healthy students from Okada, Nigeria, were tested for nasal colonization by S. aureus. Isolates were identified using standard microbiological methods. The susceptibilities of the isolates to a panel of 22 antimicrobials were tested. spa and staphylococcal cassette chromosome mec typing were performed. The prevalence of S. aureus and methicillin-resistant S. aureus (MRSA) among the students was 23% and 6%, respectively. Of the six (26.1%; 6/23) MRSA isolates detected, CC88-MRSA-IVa (n = 2) and CC7-MRSA-V (n = 2) were the most frequent clones. The CC7-MRSA-V isolates were resistant to multiple antimicrobials. Overall, resistance to beta-lactams, tetracyclines, fluoroquinolones, and aminoglycosides was detected among the S. aureus and MRSA isolates. The high prevalence of MRSA and methicillin-susceptible isolates with resistance to multiple antimicrobial classes observed among the students is an alarming finding. This study indicated the circulation of resistant clones of S. aureus in Nigerian educational institutions and the community.
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Staphylococcus aureus Resistente a Meticilina , Infecciones Estafilocócicas , Humanos , Staphylococcus aureus , Resistencia a la Meticilina , Antibacterianos/farmacología , Nigeria/epidemiología , Pruebas de Sensibilidad Microbiana , Infecciones Estafilocócicas/tratamiento farmacológico , Infecciones Estafilocócicas/epidemiología , Infecciones Estafilocócicas/microbiología , EstudiantesRESUMEN
Methicillin-resistant Staphylococcus aureus (MRSA) is a leading cause of health care-associated infections. Additionally, over the decades, the spread of community-associated (CA-MRSA) clones has become a serious problem. The aim of this study was to gain data on the current epidemiology of MRSA in Slovakia. Between January 2020 and March 2020, single-patient MRSA isolates (invasive and/or colonizing) were collected in Slovakia from hospitalized inpatients (16 hospitals) or outpatients (77 cities). Isolates were characterized via antimicrobial susceptibility testing, spa typing, SCCmec typing, the detection of mecA/mecC, genes coding for Panton-Valentine leukocidin (PVL), and the arcA gene (part of the arginine catabolic mobile element [ACME]). Out of 412 isolates, 167 and 245 originated from hospitalized patients and outpatients, respectively. Inpatients were most likely older (P < 0.001) and carried a strain exhibiting multiple resistance (P = 0.015). Isolates were frequently resistant to erythromycin (n = 320), clindamycin (n = 268), and ciprofloxacin/norfloxacin (n = 261). 55 isolates were resistant to oxacillin/cefoxitin only. By clonal structure, CC5-MRSA-II (n = 106; spa types t003, t014), CC22-MRSA-IV (n = 75; t032), and CC8-MRSA-IV (n = 65; t008) were the most frequent. We identified PVL in 72 isolates (17.48%; 17/412), with the majority belonging to CC8-MRSA-IV (n = 55; arcA+; t008, t622; the USA300 CA-MRSA clone) and CC5-MRSA-IV (n = 13; t311, t323). To the best of our knowledge, this is the first study on the epidemiology of MRSA in Slovakia. The presence of the epidemic HA-MRSA clones CC5-MRSA-II and CC22-MRSA-IV was found, as was, importantly, the emergence of the global epidemic USA300 CA-MRSA clone. The extensive spread of USA300 among inpatients and outpatients across the Slovakian regions warrants further investigation. IMPORTANCE The epidemiology of MRSA is characterized by the rise and fall of epidemic clones. Understanding the spread, as well as the evolution of successful MRSA clones, depends on the knowledge of global MRSA epidemiology. However, basic knowledge about MRSA epidemiology is still fragmented or completely missing in some parts of the world. This is the first study of MRSA epidemiology in Slovakia to identify the presence of the epidemic HA-MRSA clones CC5-MRSA-II and CC22-MRSA-IV and, importantly and unexpectedly, the emergence of the global epidemic USA300 CA-MRSA clone in the Slovakian community and hospitals. So far, USA300 has failed to spread in Europe, and this study documents an extensive spread of this epidemic clone in a European country for the first time.
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Infección Hospitalaria , Staphylococcus aureus Resistente a Meticilina , Infecciones Estafilocócicas , Humanos , Staphylococcus aureus Resistente a Meticilina/genética , Infecciones Estafilocócicas/epidemiología , Eslovaquia/epidemiología , Hospitales , Infección Hospitalaria/epidemiología , Pruebas de Sensibilidad MicrobianaRESUMEN
The Two Weeks in the World research project has resulted in a dataset of 3087 clinically relevant bacterial genomes with pertaining metadata, collected from 59 diagnostic units in 35 countries around the world during 2020. A relational database is available with metadata and summary data from selected bioinformatic analysis, such as species prediction and identification of acquired resistance genes.
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Bacterias , Genoma Bacteriano , Bacterias/genética , Biología Computacional , Bases de Datos Factuales , MetadatosRESUMEN
BACKGROUND: Colistin belongs to the last-resort antibiotics. The discovery of plasmid-bound colistin resistance mediated by the mcr-gene(s) is of great concern because, given its biological potential, there is a risk of its rapid spread. OBJECTIVES: To discuss the current literature on the methods for the screening for mcr-mediated resistance to colistin. SOURCES: Literature was drawn from a search of PubMed from 1 January 2016 to 26 April 2021. CONTENT: The selective culture-based or culture-independent approach can be used for the screening of mcr-mediated resistance to colistin in clinical samples. Rapid Polymyxin NP, Colistin Drop or Colistin Agar Spot tests are applicable for the selection of isolates with a suspected resistance to colistin that has to be confirmed by broth microdilution. The mcr-mediated resistance to colistin can be confirmed by the detection of the causal gene(s) or by phenotype using EDTA-colistin broth disc elution; production of the MCR-1 enzyme can be confirmed with lateral flow immunoassay, using matrix-assisted laser desorption/ionization time-of flight or liquid chromatography-based mass spectrometry. Whole-genome sequencing (WGS) is the ultimate typing method. When a WGS platform is not available at a healthcare facility, a WGS-outsourced service, in combination with freely available bioinformatics tools, allows for the characterization of the mcr-gene(s) carrying isolates. IMPLICATIONS: mcr-mediated colistin resistance should be monitored through active targeted screening. The broth microdilution method is required for colistin susceptibility testing but as only a selected number of clinical isolates are tested, colistin resistance, including mcr-mediated, may remain undetected. In mcr-1-positive Escherichia coli isolates, the MIC to colistin can range from 2 to 8 mg/L, so it is proposed that Enterobacterales with a colistin MIC of 2 mg/L should also be included in the mcr-mediated colistin resistance screening and those with a confirmed mcr-genotype and/or MCR-phenotype should be considered to be colistin-resistant.
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Antibacterianos , Bacterias , Colistina , Farmacorresistencia Bacteriana , Pruebas de Sensibilidad Microbiana , Antibacterianos/farmacología , Bacterias/efectos de los fármacos , Bacterias/genética , Colistina/farmacología , Farmacorresistencia Bacteriana/genética , Pruebas de Sensibilidad Microbiana/métodos , Plásmidos/genética , PolimixinasRESUMEN
Background: In order to estimate the prevalence of plasmid borne colistin resistance and to characterize in detail the mcr-positive isolates, we carried out a sentinel testing survey on the intestinal carriage of plasmid-mediated colistin-resistant Enterobacteriaceae in hospitalized patients. Methods: Between June 2018 and September 2019, 1922 faecal samples from hospitalised patients were analysed by selective culture in presence of colistin (3.5 mg/L), and in parallel by direct detection of the mcr-1 to mcr-8 genes by qPCR. The mcr-positive isolates were characterised by whole-genome sequencing. Results: The prevalence of the mcr-1 gene was 0.21% (n = 4/1922); the mcr-2 to 8 genes were not detected. The mcr-1 gene was found to be localised in the IncX4 (n = 3) and IncHI2 (n = 1) plasmid type. One Escherichia coli isolate was susceptible to colistin due to the inactivation of the mcr-1 gene through the insertion of the IS2 element; however, the colistin resistance was inducible by culture in low concentrations of colistin. One human mcr-1 positive E. coli isolate was related genetically to the mcr-1 E. coli isolate derived from turkey meat of Czech origin. Conclusions:mcr-mediated colistin resistance currently poses little threat to patients hospitalised in Czech healthcare settings. The presence of the mcr-1 gene in the human population has a possible link to domestically produced, retail meat.
RESUMEN
BACKGROUND: Travellers were recognized as a risk cohort that can be colonized by mcr-1-mediated colistin-resistant Enterobacteriaceae. We aimed to investigate the carriage of mcr-mediated colistin resistance in Enterobacteriaceae in Czech travellers or expatriates residing temporarily in the Czech Republic. METHODS: Between August 2018 and September 2019, the stool samples were cultured in enrichment broth. The enriched cultures were tested for the presence of the mcr-1-8 genes and inoculated onto selective agar with colistin. Colistin-resistant Enterobacteriaceae were tested for the presence of the mcr-1-8 genes; the mcr-positive isolates were characterised by whole genome sequencing. RESULTS: From the 177 stool samples, 15 colistin-resistant Enterobacteriaceae isolates were cultured (7.9%); two of the E. coli isolates carried the mcr-1 gene (1.1%). In the E. coli multilocus sequence type (ST) 156, the mcr-1 gene was located in an ISApl1-mcr-1-orf-ISApl1 (Tn6330) and incorporated into the chromosome; in the E. coli ST23 isolate, the mcr-1 gene was harboured by the plasmid IncX4. Both of the mcr-1 positive E. coli isolates were multidrug-resistant and one isolate was an extended-spectrum ß-lactamase producer (blaCTX-M-27). CONCLUSION: Patients with an international travel history should be monitored for the carriage of the mcr-1 gene in order to prevent its dissemination into healthcare settings.
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Colistina , Proteínas de Escherichia coli , Antibacterianos/farmacología , Cromosomas , Colistina/farmacología , Estudios Transversales , República Checa , Farmacorresistencia Bacteriana/genética , Enterobacteriaceae/genética , Escherichia coli/genética , Proteínas de Escherichia coli/genética , Humanos , Pruebas de Sensibilidad Microbiana , Plásmidos/genéticaRESUMEN
In the microbiological diagnosis of bloodstream infections (BSI), blood culture (BC) is considered the gold standard test despite its limitations such as low sensitivity and slow turnaround time. A new FDA-cleared and CE-marked platform utilizing magnetic resonance to detect amplified DNA of the six most common and/or problematic BSI pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Escherichia coli; referred to as ESKAPEc) is available and may shorten the time to diagnosis and potentially improve antimicrobial utilization. Whole blood samples from hospitalized patients with clinical signs of sepsis were analyzed using the T2Bacteria Panel (T2Biosystems) and compared to simultaneously collected BC. Discrepant results were evaluated based on clinical infection criteria, combining supporting culture results and the opinion of treating physicians. A total of 55 samples from 53 patients were evaluated. The sensitivity and specificity of the T2Bacteria panel was 94% (16 out of 17 detections of T2Bacteria-targeted organisms) and 100%, respectively, with 36.4% (8 of 22) causes of BSI detected only by this method. The T2Bacteria Panel detected pathogens on average 55 hours faster than standard BC. In our study, 9 of 15 patients with positive T2Bacteria Panel results received early-targeted antibiotic therapy and/or modification of antimicrobial treatment based on T2Bacteria Panel findings. Given the high reliability, faster time to detection, and easy workflow, the technique qualifies as a point-of-care testing approach.
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Antibacterianos/farmacología , Programas de Optimización del Uso de los Antimicrobianos/métodos , Bacteriemia/microbiología , Sangre/microbiología , Acinetobacter baumannii/efectos de los fármacos , Acinetobacter baumannii/genética , Acinetobacter baumannii/aislamiento & purificación , Bacteriemia/sangre , Bacteriemia/tratamiento farmacológico , Cultivo de Sangre , Enterococcus faecium/efectos de los fármacos , Enterococcus faecium/genética , Enterococcus faecium/aislamiento & purificación , Escherichia coli/efectos de los fármacos , Escherichia coli/genética , Escherichia coli/aislamiento & purificación , Humanos , Klebsiella pneumoniae/efectos de los fármacos , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/aislamiento & purificación , Estudios Prospectivos , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/aislamiento & purificación , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/genética , Staphylococcus aureus/aislamiento & purificaciónRESUMEN
Fast and accurate detection of causative agents of bloodstream infections remains a challenge of today's microbiology. We compared the performance of cutting-edge technology based on polymerase chain reaction coupled with electrospray ionization-mass spectrometry (PCR/ESI-MS) with that of conventional broad-range 16S rRNA PCR and blood culture to address the current diagnostic possibilities for bloodstream infections. Of 160 blood samples tested, PCR/ESI-MS revealed clinically meaningful microbiological agents in 47 samples that were missed by conventional diagnostic approaches (29.4% of all analyzed samples). Notably, PCR/ESI-MS shortened the time to positivity of the blood culture-positive samples by an average of 34 hr. PCR/ESI-MS technology substantially improved current diagnostic tools and represented an opportunity to make bloodstream infections diagnostics sensitive, accurate, and timely with a broad spectrum of microorganisms covered.
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Bacteriemia/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Reacción en Cadena de la Polimerasa/métodos , Sepsis/diagnóstico , Espectrometría de Masa por Ionización de Electrospray/métodos , Adulto , Anciano , Anciano de 80 o más Años , Bacteriemia/microbiología , Femenino , Humanos , Masculino , Técnicas Microbiológicas , Persona de Mediana Edad , ARN Ribosómico 16S , Reproducibilidad de los Resultados , Sepsis/microbiología , Adulto JovenRESUMEN
Besides natural and acquired mechanisms of resistance, bacteria can cope with presence of antibiotics by using complex mechanisms such as persistence or tolerance. The main purpose of this study was to evaluate the suitability of newly developed Tolerance Disk Test (TDtest) (Gefen et al., 2017) to detect persistent or tolerant bacterial cells in clinical isolates of Staphylococcus aureus. The principle of the test is to resuscitate the subpopulation of persistent or tolerant bacterial cells following a disk diffusion test by glucose. Results of the TDtest were evaluated using time killing experiments for three pairs of consecutive S. aureus isolates from lower respiratory airway samples of three cystic fibrosis patients with chronic staphylococcal infections. TDtest enabled semi-quantitative detection of persistent or tolerant bacterial populations in all analyzed isolates for oxacillin, vancomycin, and ciprofloxacin to which isolates studied were susceptible. Therefore, TDtest is a promising method for rapidly determining persistence/tolerance in clinical isolates of S. aureus.