RESUMEN
Carbapenem-resistance in Enterobacter spp due to acquisition of mobile carbapenemases is of concern. An Enterobacter spp grew on ChromID CARBA medium and was positive for the mCIM carbapenemase detection assay. Susceptibility testing showed resistance to aztreonam and reduced susceptibility to imipenem. Conventional PCR using FRI primers detected a blaFRI gene. Whole genome sequencing reveled a new variant; blaFRI-12 was closest in sequence to blaFRI-5 differing by 13 amino acids and was found on a unique 110Kb IncR plasmid. Given the intrinsic nature of Enterobacter spp. to be carbapenem non-susceptible, blaFRI-types may be under reported globally.
RESUMEN
The first confirmed case of COVID-19 in Quebec, Canada, occurred at Verdun Hospital on February 25, 2020. A month later, a localized outbreak was observed at this hospital. We performed tiled amplicon whole genome nanopore sequencing on nasopharyngeal swabs from all SARS-CoV-2 positive samples from 31 March to 17 April 2020 in 2 local hospitals to assess viral diversity (unknown at the time in Quebec) and potential associations with clinical outcomes. We report 264 viral genomes from 242 individuals-both staff and patients-with associated clinical features and outcomes, as well as longitudinal samples and technical replicates. Viral lineage assessment identified multiple subclades in both hospitals, with a predominant subclade in the Verdun outbreak, indicative of hospital-acquired transmission. Dimensionality reduction identified two subclades with mutations of clinical interest, namely in the Spike protein, that evaded supervised lineage assignment methods-including Pangolin and NextClade supervised lineage assignment tools. We also report that certain symptoms (headache, myalgia and sore throat) are significantly associated with favorable patient outcomes. Our findings demonstrate the strength of unsupervised, data-driven analyses whilst suggesting that caution should be used when employing supervised genomic workflows, particularly during the early stages of a pandemic.
Asunto(s)
COVID-19/virología , Infección Hospitalaria/virología , Brotes de Enfermedades , Genoma Viral/genética , SARS-CoV-2/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , COVID-19/epidemiología , COVID-19/mortalidad , Niño , Preescolar , Infección Hospitalaria/epidemiología , Brotes de Enfermedades/estadística & datos numéricos , Femenino , Haplotipos/genética , Humanos , Masculino , Persona de Mediana Edad , Filogenia , Quebec/epidemiología , SARS-CoV-2/patogenicidad , Análisis de Secuencia de ARN , Resultado del Tratamiento , Adulto JovenRESUMEN
The first confirmed case of COVID-19 in Quebec, Canada, occurred at Verdun Hospital on February 25, 2020. A month later, a localized outbreak was observed at this hospital. We performed tiled amplicon whole genome nanopore sequencing on nasopharyngeal swabs from all SARS-CoV-2 positive samples from 31 March to 17 April 2020 in 2 local hospitals to assess the viral diversity of the outbreak. We report 264 viral genomes from 242 individuals (both staff and patients) with associated clinical features and outcomes, as well as longitudinal samples, technical replicates and the first publicly disseminated SARS-CoV-2 genomes in Quebec. Viral lineage assessment identified multiple subclades in both hospitals, with a predominant subclade in the Verdun outbreak, indicative of hospital-acquired transmission. Dimensionality reduction identified two subclades that evaded supervised lineage assignment methods, including Pangolin, and identified certain symptoms (headache, myalgia and sore throat) that are significantly associated with favorable patient outcomes. We also address certain limitations of standard SARS-CoV-2 bioinformatics procedures, notably when presented with multiple viral haplotypes.
RESUMEN
The accurate laboratory detection of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a crucial element in the fight against coronavirus disease 2019 (COVID-19). Reverse transcription-polymerase chain reaction testing on combined oral and nasopharyngeal swab (ONPS) suffers from several limitations, including the need for qualified personnel, the discomfort caused by invasive nasopharyngeal sample collection, and the possibility of swab and transport media shortage. Testing on saliva would represent an advancement. The aim of this study was to compare the concordance between saliva samples and ONPS for the detection of SARS-CoV-2 on various commercial and laboratory-developed tests (LDT). Individuals were recruited from eight institutions in Quebec, Canada, if they had SARS-CoV-2 RNA detected on a recently collected ONPS, and accepted to provide another ONPS, paired with saliva. Assays available in the different laboratories (Abbott RealTime SARS-CoV-2, Cobas® SARS-CoV-2, Simplexa™ COVID-19 Direct, Allplex™ 2019-nCoV, RIDA®GENE SARS-CoV-2, and an LDT preceded by three different extraction methods) were used to determine the concordance between saliva and ONPS results. Overall, 320 tests were run from a total of 125 saliva and ONPS sample pairs. All assays yielded similar sensitivity when saliva was compared to ONPS, with the exception of one LDT (67% vs. 93%). The mean difference in cycle threshold (∆C t ) was generally (but not significantly) in favor of the ONPS for all nucleic acid amplification tests. The maximum mean ∆âââââC t was 2.0, while individual ∆C t varied importantly from -17.5 to 12.4. Saliva seems to be associated with sensitivity similar to ONPS for the detection of SARS-CoV-2 by various assays.
Asunto(s)
Prueba de Ácido Nucleico para COVID-19/normas , COVID-19/diagnóstico , Pruebas Diagnósticas de Rutina/normas , ARN Viral/genética , SARS-CoV-2/genética , COVID-19/epidemiología , COVID-19/virología , Prueba de Ácido Nucleico para COVID-19/instrumentación , Prueba de Ácido Nucleico para COVID-19/métodos , Pruebas Diagnósticas de Rutina/instrumentación , Pruebas Diagnósticas de Rutina/métodos , Humanos , Boca/virología , Nasofaringe/virología , Quebec/epidemiología , Saliva/virología , Sensibilidad y Especificidad , Manejo de Especímenes/normasRESUMEN
OBJECTIVE: To determine the frequency with which Clostridium difficile was detected in stool specimens from outpatients and patients hospitalized for less than 4 days to assess the usefulness of routine laboratory screening for detecting this enteric pathogen. METHODS: Seven hundred and forty-one specimens from 398 patients were cultured over a 6-month period for Salmonella, Shigella, Yersinia, Escherichia coli O157:H7, Campylobacter and Clostridium difficile. Clostridium difficile culture-positive samples were further tested for cytotoxin production. RESULTS: Campylobacter, Salmonella, Shigella and E. coli O157:H7 were isolated in 50 (6.7%) specimens from 35 (8.8%) patients. Clostridium difficile was cultured from 88 (11.9%) specimens from 35 (8.8%) patients and its cytotoxin detected in 35 (4.7%) specimens of 12 (3%) patients. Clostridium difficile was the second most frequent enteric pathogen after Campylobacter. Of 178 (24%) specimens submitted with a specific request for Clostridium difficile testing, 13 (7.3%) were cytotoxin positive (three patients); of 563 specimens for which Clostridium difficile was not requested, 22 (3.9%) were cytotoxin positive (nine patients). CONCLUSIONS: Nine of 12 patients with cytotoxin-positive specimens would have gone undiagnosed in the laboratory had all stool samples submitted not been tested. These results suggest that Clostridium difficile disease is under-recognized and that testing all stool samples for Clostridium difficile may be warranted in our community of patients.