Rapid Detection of Carbapenemase and Extended-Spectrum ß-Lactamase Producing Gram-Negative Bacteria Directly from Positive Blood Cultures Using a Novel Protocol.

BACKGROUND: Early and adequate antibiotic treatment is the cornerstone of improving clinical outcomes in patients with bloodstream infections (BSI). Delays in appropriate antimicrobial therapy have catastrophic consequences for patients with BSI. Microbiological characterization of multi-drug resistant pathogens (MDRP) allows clinicians to provide appropriate treatments. Current microbiologic techniques may take up to 96 h to identify causative pathogens and their resistant patterns. Therefore, there is an important need to develop rapid diagnostic strategies for MDRP. We tested a modified protocol to detect carbapenemase and extended-spectrum ß-lactamase (ESBL) producing Gram-negative bacteria (GNB) from positive blood cultures. METHODS: This is a prospective cohort study of consecutive patients with bacteremia. We developed a modified protocol using the HB&L® system to detect MDRP. The operational characteristics were analyzed for each test (HB&L-ESBL/AmpC® and HB&L-Carbapenemase® kits). The kappa coefficient, sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), likelihood ratios (LR) with 95% confidence intervals (CI), and reduction in identification time of this novel method were calculated. RESULTS: Ninety-six patients with BSI were included in the study. A total of 161 positive blood cultures were analyzed. Escherichia coli (50%, 81/161) was the most frequently identified pathogen, followed by Klebsiella pneumoniae (15%, 24/161) and Pseudomonas aeruginosa (8%, 13/161). Thirty-three percent of isolations had usual resistance patterns. However, 34/161 (21%) of identified pathogens were producers of carbapenemases and 21/161 (13%) of extended-spectrum ß-lactamases. Concordance between our HB&L® modified protocol and the traditional method was 99% (159/161). Finally, identification times were significantly shorter using our HB&L®-modified protocol than traditional methods: median (IQR) 19 h (18, 22) vs. 61 h (60, 64), p < 0.001. CONCLUSIONS: Here, we provide novel evidence that using our HB&L®-modified protocol is an effective strategy to reduce the time to detect MDRP producers of carbapenemases or extended-spectrum ß-lactamases, with an excellent concordance rate when compared to the gold standard. Further studies are needed to confirm these findings and to determine whether this method may improve clinical outcomes.

Cerebral phaeohyphomycosis caused by Alternaria spp.: A case report.

Phaeohyphomycosis is a group of infections caused by pigmented, black, dematiaceous fungi and is responsible for cutaneous, superficial and deep mycoses, disseminated infection and brain abscesses. The primary agents involved include Alternaria spp., Exophiala spp. and Cladophialophora spp. Invasive systemic presentation is rare and in most cases is associated with immunosuppression; for this reason, reported cases of Alternaria spp. infection are scarce. This report describes the case of a 66-year-old man with a history of renal transplantation from a cadaveric donor 1 year ago, which was considered as the primary risk factor. The characteristics of the infection, procedures performed, microbiological findings and treatment provided are described.

[Evaluation of three screening methods for detection of carbapenemase-producing Enterobacteriaceae in rectal swabs]. / Evaluación de tres métodos de tamizaje para detección de Enterobacteriaceae productoras de carbapenemasas en hisopados rectales.

BACKGROUND: Carbapenemase-producing Enterobacteriaceae (CPE) have taken great importance on public health at global scale, which makes it necessary to implement rapid test for its prompt detection. AIM: To evaluate three screening methods to detect CPE in rectal swabs. MATERIAL AND METHODS: Transverse study, prospective. Seventy three rectal swabs were evaluated by three methodologies. Microorganism identification and susceptibility testing were made using automated systems. Carbapenemase production was confirmed by modified Hodge test and synergy tests using boronic acid and EDTA. RESULTS: The method 1 (ChromID CARBA®) detected 20 positive samples (27.4%), 5 false positives (6.9 %), with concordance index of 93.2%, sensitivity 100% and specificity of 90%. Method 2 (HB&L Carbapenemase®) detected 17 positive samples (23.3%) and 3 false negatives (4.1%). The sensitivity and specificity of the assay were 85% and 100%, with concordance index of 95.9%. Method 3 (Xpert Carba-R®) detected 19 positive samples (57.5%) and 1 false negatives (3.1%), sensitivity 95%, specificity 100% and concordance index of 97%. DISCUSSION: There is a wide variety of methodologies for rapid detection of carbapenemase-producing microorganisms. Choosing the best method must have as requirement a good sensitivity, specificity, and cost-effectiveness.

Evaluación de tres métodos de tamizaje para detección de Enterobacteriaceae productoras de carbapenemasas en hisopados rectales / Evaluation of three screening methods for detection of carbapenemase-producing Enterobacteriaceae in rectal swabs

Resumen Introducción: Las Enterobacteriaceae productoras de carbapenemasas (EPC) han tomado gran importancia en salud pública a una escala global, haciendo necesario implementar test rápidos para su detección oportuna. Objetivo: Evaluar tres metodologías para el tamizaje de EPC en hisopados rectales. Materiales y Métodos: Estudio prospectivo transversal. Se evaluaron 73 hisopados rectales por tres metodologías. Se realizó identificación y evaluación de susceptibilidad por sistemas automatizados y la producción de carbapenemasas se confirmó por test de Hodge modificado, sinergia con ácido borónico y EDTA. Resultados: Método 1 (ChromID CARBA®): detectó 20 muestras positivas (27,4%), 5 falsos positivos (6,9%), con índice de concordancia de 93,2%, sensibilidad 100% y especificidad de 90%. Método 2 (HB&L Carbapenemase®): detectó 17 muestras positivas (23,3%) y 3 falsos negativos (4,1%). La sensibilidad y especificidad fue 85 y 100% respectivamente, con concordancia de 95,9%. Método 3 (Xpert Carba-R®): detectó 19 muestras positivas (57,5 %) y 1 falso negativo (3,1%), sensibilidad 95%, especificidad 100% e índice de concordancia de 97%. Discusión: Existe amplia variedad de metodologías para búsqueda y detección rápida de microorganismos productores de carbapenemasas. La elección del método debe tener como requisito una buena sensibilidad y especificidad, rapidez y costo efectividad.

Background: Carbapenemase-producing Enterobacteriaceae (CPE) have taken great importance on public health at global scale, which makes it necessary to implement rapid test for its prompt detection. Aim: To evaluate three screening methods to detect CPE in rectal swabs. Material and Methods: Transverse study, prospective. Seventy three rectal swabs were evaluated by three methodologies. Microorganism identification and susceptibility testing were made using automated systems. Carbapenemase production was confirmed by modified Hodge test and synergy tests using boronic acid and EDTA. Results: The method 1 (ChromID CARBA®) detected 20 positive samples (27.4%), 5 false positives (6.9 %), with concordance index of 93.2%, sensitivity 100% and specificity of 90%. Method 2 (HB&L Carbapenemase®) detected 17 positive samples (23.3%) and 3 false negatives (4.1%). The sensitivity and specificity of the assay were 85% and 100%, with concordance index of 95.9%. Method 3 (Xpert Carba-R®) detected 19 positive samples (57.5%) and 1 false negatives (3.1%), sensitivity 95%, specificity 100% and concordance index of 97%. Discussion: There is a wide variety of methodologies for rapid detection of carbapenemase-producing microorganisms. Choosing the best method must have as requirement a good sensitivity, specificity, and cost-effectiveness.

Características de la infección por Clostridium difficile en una institución de alta complejidad y reporte de la circulación en Colombia de la cepa hipervirulenta NAP1/027 / Characteristics of Clostridium difficile infection in a high complexity hospital and report of the circulation of the NAP1/027 hypervirulent strain in Colombia

Resumen Introducción. Clostridium difficile es el principal responsable de la diarrea asociada al uso de antibióticos. En Colombia y en Latinoamérica, el conocimiento sobre el comportamiento epidemiológico de la infección por C. difficile todavía es limitado. Objetivo. Describir las características de una serie de pacientes con infección por C.difficile . Materiales y métodos. Se hizo un estudio descriptivo de una serie de casos de pacientes con infección por C. difficile atendidos en la Fundación Clínica Shaio, entre enero de 2012 y noviembre de 2015. Resultados. Se estudiaron 36 pacientes con una edad promedio de 65 años. Se determinaron los siguientes factores relacionados con la infección por C. difficile: uso previo de antimicrobianos (94,4 %), hospitalización en los últimos tres meses (66,7 %) y uso de inhibidores de la bomba de protones (50 %). Las comorbilidades más comunes fueron la enfermedad renal crónica (41,7 %) y la diabetes mellitus (30,6 %). Los síntomas más frecuentes fueron más de tres deposiciones diarreicas (97,1 %) y dolor abdominal (42,9 %). En cuanto a la gravedad de los casos, 44,4 % se clasificó como leve a moderado, 38,9 % como grave, y 11,1 % como complicado o grave. El método de diagnóstico más utilizado (63,8% de los pacientes) fue la identificación de la toxina mediante reacción en cadena de la polimerasa (PCR). La mortalidad global durante la hospitalización fue de 8 %. Se identificaron cuatro cepas del serotipo NAP1/027 y nueve muestras fueron positivas para la toxina binaria. Conclusión. La infección por C. difficile debe sospecharse en pacientes con deposiciones diarreicas y factores asociados tradicionalmente a esta enfermedad. Se reportó la circulación de cepas hipervirulentas del serotipo NAP1/027 en Colombia, lo cual debe enfrentarse con la vigilancia epidemiológica y el diagnóstico temprano.

Abstract Introduction: Clostridium difficile is the main pathogen related to healthcare-associated diarrhea and it is the cause of 20 to 30% of diarrhea cases caused by antibiotics. In Colombia and Latin America, the knowledge about the epidemiological behavior of this infection is limited. Objective: To describe the characteristics of a series of patients with C. difficile infection. Materials and methods: We performed a descriptive case series study of patients with C. difficile infection hospitalized in the Fundación Clínica Shaio from January, 2012, to November, 2015. Results: We analyzed 36 patients. The average age was 65 years. The risk factors associated with the infection were: previous use of antibiotics (94.4%), prior hospitalization in the last three months (66.7%) and use of proton pump inhibitors (50%). The most common comorbidities were chronic kidney disease (41.7%) and diabetes mellitus (30.6%). The most frequent symptoms were more than three loose stools per day (97.1%) and abdominal pain (42.9%). According to the severity of the disease, 44.4% of cases were classified as mild to moderate, 38.9% as severe, and 11.1% as complicated or severe. The detection of the toxin by PCR (GeneXpert) was the most common diagnostic procedure (63.8%). Global mortality during hospitalization was 8%. We identified four strains with serotype NAP1/027 and nine samples positive for binary toxin. Conclusion: Clostridium difficile infection should be suspected in patients with diarrhea and traditional risk factors associated with this disease. We report the circulation of the hypervirulent strain serotype NAP1/027 in Colombia, which should be countered with epidemiological surveillance and a prompt diagnosis.

[Characteristics of Clostridium difficile infection in a high complexity hospital and report of the circulation of the NAP1/027 hypervirulent strain in Colombia].

INTRODUCTION: Clostridium difficile is the main pathogen related to healthcare-associated diarrhea and it is the cause of 20 to 30% of diarrhea cases caused by antibiotics. In Colombia and Latin America, the knowledge about the epidemiological behavior of this infection is limited. OBJECTIVE: To describe the characteristics of a series of patients with C. difficile infection. MATERIALS AND METHODS: We performed a descriptive case series study of patients with C. difficile infection hospitalized in the Fundación Clínica Shaio from January, 2012, to November, 2015. RESULTS: We analyzed 36 patients. The average age was 65 years. The risk factors associated with the infection were: previous use of antibiotics (94.4%), prior hospitalization in the last three months (66.7%) and use of proton pump inhibitors (50%). The most common comorbidities were chronic kidney disease (41.7%) and diabetes mellitus (30.6%). The most frequent symptoms were more than three loose stools per day (97.1%) and abdominal pain (42.9%). According to the severity of the disease, 44.4% of cases were classified as mild to moderate, 38.9% as severe, and 11.1% as complicated or severe. The detection of the toxin by PCR (GeneXpert) was the most common diagnostic procedure (63.8%). Global mortality during hospitalization was 8%. We identified four strains with serotype NAP1/027 and nine samples positive for binary toxin. CONCLUSION: Clostridium difficile infection should be suspected in patients with diarrhea and traditional risk factors associated with this disease. We report the circulation of the hypervirulent strain serotype NAP1/027 in Colombia, which should be countered with epidemiological surveillance and a prompt diagnosis.