RESUMO
In this multisite study, Vitek 2 AST-Gram-Negative Ceftazidime/Avibactam test results for 1,073 isolates (866 Enterobacterales and 207 Pseudomonas aeruginosa) were compared to the Clinical and Laboratory Standards Institute (CLSI) broth microdilution (BMD) reference method. The results were analyzed for essential agreement (EA), category agreement (CA), major error rates, and very major error rates following FDA/ISO performance criteria using the FDA-recognized CLSI/EUCAST breakpoints (sensitive [S], ≤8/4 µg/ml; resistant [R], ≥16/4 µg/ml). The overall EA was 94.5% (1,014/1,073) and CA was 98.7% (1,059/1,073). No very major errors were reported. The major error rate was 1.4% (14/998). Out of 14 major errors, 9 were within EA. Based on the EA and lack of an intermediate category for ceftazidime-avibactam (CZA), the adjusted major error rate for FDA criteria was 0.5% (5/998). The performance for ISO criteria after error resolutions included EA of 94.5% (1,014/1,073), CA of 98.9% (1,061/1,073), major error of 1.2% (12/998), and no very major error. Vitek 2 met the ISO and FDA criteria of ≥95% reproducibility and ≥95% quality control (QC) results within acceptable ranges for QC organisms. Vitek 2 overall performance for Enterobacterales and P. aeruginosa met or exceeded the FDA and ISO performance criteria; thus, it is a reliable alternative to the BMD reference method for routine CZA susceptibility testing.
Assuntos
Ceftazidima , Pseudomonas aeruginosa , Antibacterianos/farmacologia , Compostos Azabicíclicos , Ceftazidima/farmacologia , Combinação de Medicamentos , Enterobacteriaceae , Humanos , Testes de Sensibilidade Microbiana , Reprodutibilidade dos TestesRESUMO
The diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has ramifications on both an individual level and a public health level. The use of appropriate testing mechanisms is paramount to preventing transmission, along with offering treatment to those who are infected and show appropriate symptomatology. The choice of employing a specific test often relies on laboratory capabilities, including the abilities of the medical technologists, the cost of testing platforms, and the individual quirks of each test. This chapter intends to discuss the relevant issues relating to diagnostic testing for SARS-CoV-2, including specimen types and collection methods, viral detection methods, and serological testing.
Assuntos
COVID-19 , Teste para COVID-19 , Técnicas de Laboratório Clínico , Testes Diagnósticos de Rotina , Humanos , SARS-CoV-2RESUMO
Nontoxigenic Clostridium difficile (NTCD) has been shown to prevent fatal C. difficile infection in the hamster model when hamsters are challenged with standard toxigenic C. difficile strains. The purpose of this study was to determine if NTCD can prevent C. difficile infection in the hamster model when hamsters are challenged with restriction endonuclease analysis group BI C. difficile strains. Groups of 10 hamsters were given oral clindamycin, followed on day 2 by 10(6) CFU of spores of NTCD strain M3 or T7, and were challenged on day 5 with 100 CFU of spores of BI1 or BI6. To conserve animals, results for control hamsters challenged with BI1 or BI6 from the present study and controls from previous identical experiments were combined for statistical comparisons. NTCD strains M3 and T7 achieved 100% colonization and were 100% protective against challenge with BI1 (P ≤ 0.001). M3 colonized 9/10 hamsters and protected against BI6 challenge in the colonized hamsters (P = 0.0003). T7 colonized 10/10 hamsters, but following BI6 challenge, cocolonization occurred in 5 hamsters, 4 of which died, for protection of 6/10 animals (P = 0.02). NTCD colonization provides protection against challenge with toxigenic BI group strains. M3 is more effective than T7 in preventing C. difficile infection caused by the BI6 epidemic strain. Prevention of C. difficile infection caused by the epidemic BI6 strain may be more challenging than that of infections caused by historic BI1 and non-BI C. difficile strains.
Assuntos
Clostridioides difficile/imunologia , Proteção Cruzada , Enterocolite Pseudomembranosa/prevenção & controle , Enterocolite Pseudomembranosa/veterinária , Imunidade Heteróloga , Esporos Bacterianos/imunologia , Imunidade Adaptativa , Animais , Antibacterianos/farmacologia , Clindamicina/farmacologia , Clostridioides difficile/efeitos dos fármacos , Clostridioides difficile/patogenicidade , Cricetinae , Enterocolite Pseudomembranosa/imunologia , Enterocolite Pseudomembranosa/microbiologia , Masculino , Mesocricetus , Esporos Bacterianos/efeitos dos fármacos , Esporos Bacterianos/patogenicidadeRESUMO
Toxin A has historically been regarded as the primary virulence determinant in Clostridium difficile infection, but naturally occurring toxin A-negative, toxin B-positive (A-/B+) C. difficile strains are known to be virulent. To determine the role of toxin B in these strains, we immunized hamsters with a toxoid prepared from purified toxin B to determine whether they would be protected from lethal challenge with an A-/B+ strain of C. difficile.
Assuntos
Proteínas de Bactérias/metabolismo , Toxinas Bacterianas/metabolismo , Clostridioides difficile/imunologia , Infecções por Clostridium/imunologia , Enterotoxinas/metabolismo , Toxoides/imunologia , Vacinação , Fatores de Virulência/metabolismo , Animais , Clostridioides difficile/metabolismo , Clostridioides difficile/patogenicidade , Infecções por Clostridium/microbiologia , Cricetinae , Toxoides/administração & dosagemRESUMO
BACKGROUND: An epidemic strain of Clostridium difficile designated by restriction endonuclease analysis (REA) as group BI has caused multiple outbreaks of severe C. difficile infection (CDI). The treatment response of patients infected with this strain is uncertain. METHODS: Clostridium difficile isolates were collected from 2 phase 3 clinical trials comparing fidaxomicin to vancomycin and typed using REA. Clinical cure and recurrence outcomes were analyzed by strain type of the infecting organism, BI and non-BI, using both univariate and multivariate analyses. RESULTS: From 999 patients, 719 isolates were available for typing (356 fidaxomicin treated and 363 vancomycin treated). BI was the most common REA group (34% of isolates). Patients infected with BI had lower cure rates (86.6%; 214 of 247) than those infected with non-BI strains (94.3%; 445 of 472) (P < .001). The cure rate difference between the BI and non-BI patients was significant for both vancomycin (P = .02) and fidaxomicin (P = .007). BI patients had a recurrence rate of 27.4% (51 of 186), compared with a recurrence rate of 16.6% (66 of 397) in non-BI patients (P = .002). By multivariate analysis, BI infection was statistically significant as a risk factor for reduced cure (odds ratio [OR], 0.48; 95% confidence interval [CI], .27-.85; P = .030) and for increased recurrence (OR, 1.57; 95% CI, 1.01-2.45; P = .046). CONCLUSIONS: The clinical cure rate of patients infected with the epidemic BI C. difficile strain is lower than the cure rate of those infected with non-BI strains whether treated with fidaxomicin or vancomycin. Similarly, the CDI recurrence rate is increased in patients with the BI strain compared with patients with other C. difficile strains.
Assuntos
Clostridioides difficile/patogenicidade , Infecções por Clostridium/tratamento farmacológico , Infecções por Clostridium/microbiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Aminoglicosídeos/uso terapêutico , Antibacterianos , Clostridioides difficile/classificação , Clostridioides difficile/genética , Clostridioides difficile/isolamento & purificação , Infecções por Clostridium/epidemiologia , Método Duplo-Cego , Feminino , Fidaxomicina , Humanos , Masculino , Pessoa de Meia-Idade , Tipagem Molecular , Polimorfismo de Fragmento de Restrição , Proibitinas , Recidiva , Resultado do Tratamento , Vancomicina/uso terapêuticoRESUMO
BACKGROUND: The recent epidemic of Clostridium difficile infection (CDI) has been attributed largely to the REA strain group BI (aka NAP1/027). Current isolates of the BI group differ from historic non-epidemic BI strains in that they have developed resistance to the newer fluoroquinolone (FQ), moxifloxacin. The acquisition of moxifloxacin resistance has been associated with the rise in frequency of CDI caused by these isolates. In order to explore the effect of FQ resistance on CDI we compared colonization and mortality in hamsters challenged with a historic (BI1 - susceptible to moxifloxacin) and a recent epidemic (BI17 - resistant to moxifloxacin) BI strain of CD during continuous administration of 3 different FQs. METHODS: Groups of 6 hamsters were treated with a 5 day course of ciprofloxacin, levofloxacin, or moxifloxacin given orally once per day. Each hamster was then challenged with 1 × 10(4) CFU of either BI1 or BI17 on day 3 or day 5. Colonization and time to death were recorded. RESULTS: Colonization efficiency (CE) following moxifloxacin (92%) was significantly greater than following levofloxacin (50%, p < 0.01) or ciprofloxacin (42%, p < 0.01) for both strains combined. The CE of BI17 was higher than BI1 for ciprofloxacin (67% vs 17%, p = 0.04) and levofloxacin (83% vs 17%, p < 0.01), but not moxifloxacin (100% and 83%, p = 0.48) administration. BI17 also showed a shorter time from inoculation to death than BI1 following moxifloxacin administration (1.8 days vs 3.9 days, p < 0.01). Moxifloxacin shortened the time from inoculation to death compared to ciprofloxacin in hamsters challenged with BI17 (1.8 days vs 4.0 days, p < 0.01) but not levofloxacin (1.8 days vs 2.0 days, p = NS). CONCLUSIONS: For the epidemic BI17 strain, ciprofloxacin, levofloxacin and moxifloxacin have similar colonization rates, suggesting that the acquisition of high-level FQ resistance increases colonization rates in association with any FQ. Historic strain BI1 which does not carry high-level FQ resistance colonized efficiently only in the presence of moxifloxacin, possibly explaining lower rates of CDI historically prior to the widespread clinical use of moxifloxacin (and gatifloxacin). Current high rates and severity of CDI from 2000 to 2010 may in part be associated with the acquisition of high-level FQ resistance in BI strains and higher patient exposure rates of all FQs, especially moxifloxacin.
Assuntos
Antibacterianos/administração & dosagem , Clostridioides difficile/patogenicidade , Infecções por Clostridium/prevenção & controle , Fluoroquinolonas/administração & dosagem , Animais , Compostos Aza/administração & dosagem , Ciprofloxacina/administração & dosagem , Clostridioides difficile/isolamento & purificação , Infecções por Clostridium/microbiologia , Cricetinae , Farmacorresistência Bacteriana , Humanos , Levofloxacino , Masculino , Mesocricetus , Testes de Sensibilidade Microbiana/métodos , Moxifloxacina , Ofloxacino/administração & dosagem , Proibitinas , Quinolinas/administração & dosagemRESUMO
BACKGROUND: The timing of and risk factors for intestinal colonization with multidrug-resistant Enterobacteriaceae (MDRE) are still poorly understood in areas with high MDRE carriage. We determined the prevalence, timing, and risk factors associated with MDRE intestinal colonization among infants in southern Sri Lanka. METHODS: Women and their newborn children were enrolled within 48 h after delivery in southern Sri Lanka. Rectal swabs were collected from women and infants at enrollment and 4-6 weeks later. Enterobacteriaceae were isolated and identified as MDRE (positive for extended-spectrum ß-lactamases or carbapenem resistant) using standard microbiologic procedures. We used exact methods (Fisher's exact and Kruskal-Wallis tests) and multivariable logistic regression to identify sociodemographic and clinical features associated with MDRE intestinal colonization. Whole-genome sequencing was performed on selected MDRE isolates to identify phylogroups and antibiotic resistance-encoding genes were identified with NCBI's AMRfinder tool. RESULTS: Overall, 199 post-partum women and 199 infants were enrolled; 148/199 (74.4%) women and 151/199 (75.9%) infants were reassessed later in the community. Twenty-four/199 (12.1%) women and 3/199 (1.5%) infants displayed intestinal colonization with MDRE at enrollment, while 26/148 (17.6%) women and 24/151 (15.9%) infants displayed intestinal colonization with MDRE at the reassessment. While there were no risk factors associated with infant colonization at enrollment, multivariable analysis indicated that risk factors for infant colonization at reassessment included mother colonized at enrollment (aOR = 3.62) or reassessment (aOR = 4.44), delivery by Cesarean section (aOR = 2.91), and low birth weight (aOR = 5.39). Of the 20 MDRE isolates from infants that were sequenced, multilocus sequence typing revealed that 6/20 (30%) were clustered on the same branch as MDRE isolates found in the respective mothers. All sequenced isolates for mothers (47) and infants (20) had at least one ESBL-producing gene. Genes encoding fosfomycin resistance were found in 33/47 (70%) of mothers' isolates and 16/20 (80%) of infants' isolates and genes encoding resistance to colistin were found in one (2%) mother's isolate. CONCLUSIONS: Our results suggest that a substantial proportion of infants undergo MDRE intestinal colonization within 6 weeks of birth, potentially due to postnatal rather than intranatal transmission.
Assuntos
Farmacorresistência Bacteriana Múltipla , Infecções por Enterobacteriaceae/epidemiologia , Adulto , Enterobacteriaceae/efeitos dos fármacos , Feminino , Hospitais de Ensino , Humanos , Recém-Nascido , Masculino , Gravidez , Reto/microbiologia , Sri Lanka/epidemiologia , Sequenciamento Completo do GenomaRESUMO
Cutavirus is a new member of the Parvoviridae family. It was first discovered in 2016 through unbiased metagenomics performed on fecal samples collected from patients with diarrhea, and also in skin biopsies collected from patients with cutaneous T-cell lymphoma (CTCL, also known as mycosis fungoides). We have systematically reviewed the literature to describe the discovery, genomic organization, prevalence, and geographic distribution of cutavirus.
Assuntos
Parvovirus/classificação , Parvovirus/genética , Biópsia , Diarreia/epidemiologia , Diarreia/etiologia , Variação Genética , Genoma Viral , Humanos , Linfoma Cutâneo de Células T/epidemiologia , Linfoma Cutâneo de Células T/etiologia , Metagenoma , Metagenômica/métodos , Epidemiologia Molecular , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/virologia , Parvovirus/isolamento & purificação , Estudos SoroepidemiológicosRESUMO
Clostridium difficile is a widely distributed pathogen with multiple strain types as determined by restriction endonuclease analysis (REA) and by PCR ribotyping, two well-characterized typing systems. In this study, REA typing was performed on 894C. difficile isolates from patients enrolled from 16 countries on three continents in two large, recently conducted clinical treatment trials of C. difficile infection. REA group BI (Ribotype 027) isolates were the most common strains identified and were widely distributed throughout North America, but restricted to three of thirteen countries in Europe. REA group J (Ribotype 001) isolates were the most common strains identified in Europe and non-specific REA groups (historically less frequent) were the most common strains identified in Australia. REA groups BI, J, G and CF correlated with specific PCR ribotypes whereas more than one ribotype was found within REA groups Y, BK, and K. International surveillance of C. difficile strains is important to document the changing epidemiology of this enteric pathogen that continues to cause healthcare facility outbreaks and sporadic infections in other settings.
Assuntos
Clostridioides difficile/classificação , Clostridioides difficile/genética , Enzimas de Restrição do DNA , Enterocolite Pseudomembranosa/epidemiologia , Ribotipagem , Austrália/epidemiologia , Técnicas de Tipagem Bacteriana , Clostridioides difficile/isolamento & purificação , Enterocolite Pseudomembranosa/microbiologia , Europa (Continente)/epidemiologia , Humanos , América do Norte/epidemiologia , Reação em Cadeia da Polimerase , ProibitinasRESUMO
A point-prevalence study of antimicrobial use among inpatients at 5 public hospitals in Sri Lanka revealed that 54.6% were receiving antimicrobials: 43.1% in medical wards, 68.0% in surgical wards, and 97.6% in intensive care wards. Amoxicillin-clavulanate was most commonly used for major indications. Among patients receiving antimicrobials, 31.0% received potentially inappropriate therapy.
Assuntos
Anti-Infecciosos/uso terapêutico , Unidades Hospitalares/estatística & dados numéricos , Hospitais Públicos/estatística & dados numéricos , Prescrição Inadequada/estatística & dados numéricos , Adulto , Gestão de Antimicrobianos , Estudos Transversais , Resistência Microbiana a Medicamentos , Feminino , Fidelidade a Diretrizes , Pesquisas sobre Atenção à Saúde , Unidades Hospitalares/classificação , Hospitalização , Humanos , Masculino , Pessoa de Meia-Idade , Sri Lanka , Adulto JovemRESUMO
BACKGROUND: North American and European hospitals have reported outbreaks of Clostridium difficile-associated disease with unexpectedly high mortality caused by a newly recognized group of C. difficile strains, group BI. Our objective was to compare, in hamsters, the virulence of a historical nonepidemic BI type, BI1, with that of 2 recent epidemic BI types, BI6 and BI17, and with that of 2 standard toxigenic strains, K14 and 630. METHODS: For each strain, 10 hamsters were given 1 dose of clindamycin, followed 5 days later with 100 C. difficile spores administered by gastric inoculation. Outcomes were recorded. RESULTS: The hamster model demonstrated variations in mean times from inoculation to death (for BI6, 40 h; for BI1, 48 h; for K14, 49 h; for BI17, 69 h; for 630, 102 h; for BI6, BI1, and K14 vs. 630, P< .01; for BI17 vs. 630, P< .05) and from colonization to death (for BI1, 7 h; for BI17, 13 h; for BI6, 16 h; for K14, 17 h; for 630, 52 h; for BI1, BI17, BI6, and K14 vs. 630, P< .01). CONCLUSION: Group BI strains were not more rapidly fatal than the standard toxinotype 0 strain K14 but were more rapidly fatal than the standard toxinotype 0 strain 630. BI6, the most common BI type in our collection, was particularly virulent in hamsters, consistently causing death within 48 h of inoculation.