Interhospital Spread of blaVIM-1- and blaCTX-M-15-Producing K. pneumoniae ST15 on an IncR Plasmid in Southern Spain.

In 2014-2015, the main CTX-M-15- and OXA-48-producing clone in our region was ST15. Recently, K. pneumoniae ST15 isolates co-producing VIM-1 and CTX-M-15 were detected in several hospitals. The aim was to study the emergence and acquisition of this carbapenemase. Between 2017 and 2019, four hospitals submitted twenty-nine VIM-1- and CTX-M-15-producing K. pneumoniae ST15 isolates to our laboratory. Seven representatives of each XbaI PFGE pulsotype were sequenced using short- and long-read technologies. RAST, CGE databases, and Pathogenwatch were used for resistance determinants and capsule-type analysis. Plasmid comparison was performed with Easyfig2.1. Phylogenetic analysis included other contemporary ST15 isolates from Spain. The 29 isolates were clustered into seven different pulsotypes. The selected genomes, from three hospitals in two different provinces, were clustered together (fewer than 35 alleles) and differed by more than 100 alleles from other ST15 isolates obtained in the region. These seven isolates harbored one IncR plasmid (200-220 kb) with a common backbone and four regions flanked by IS26: one contained blaVIM-1, another contained blaCTX-M-15, the third contained blaOXA-1, and the fourth harbored heavy-metal-tolerance genes. The two initial plasmids, from two different centers, were identical, and rearrangement of four regions was observed in the five subsequent plasmids. Our findings showed the first intercenter dissemination of IncR plasmids carrying blaVIM-1, blaCTX-M-15, and metal-tolerance genes mediated by a new lineage of K. pneumoniae ST15. Two different capture events of the blaVIM-1 gene or different IS26-mediated plasmid rearrangements from a common ancestor may explain plasmid variations.

The CARBA-MAP study: national mapping of carbapenemases in Spain (2014-2018).

Introduction: Infections caused by carbapenem-resistant Enterobacterales (CRE) and carbapenem-resistant Pseudomonas aeruginosa, including isolates producing acquired carbapenemases, constitute a prevalent health problem worldwide. The primary objective of this study was to determine the distribution of the different carbapenemases among carbapenemase-producing Enterobacterales (CPE, specifically Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae complex, and Klebsiella aerogenes) and carbapenemase-producing P. aeruginosa (CPPA) in Spain from January 2014 to December 2018. Methods: A national, retrospective, cross-sectional multicenter study was performed. The study included the first isolate per patient and year obtained from clinical samples and obtained for diagnosis of infection in hospitalized patients. A structured questionnaire was completed by the participating centers using the REDCap platform, and results were analyzed using IBM SPSS Statistics 29.0.0. Results: A total of 2,704 carbapenemase-producing microorganisms were included, for which the type of carbapenemase was determined in 2692 cases: 2280 CPE (84.7%) and 412 CPPA (15.3%), most often using molecular methods and immunochromatographic assays. Globally, the most frequent types of carbapenemase in Enterobacterales and P. aeruginosa were OXA-48-like, alone or in combination with other enzymes (1,523 cases, 66.8%) and VIM (365 cases, 88.6%), respectively. Among Enterobacterales, carbapenemase-producing K. pneumoniae was reported in 1821 cases (79.9%), followed by E. cloacae complex in 334 cases (14.6%). In Enterobacterales, KPC is mainly present in the South and South-East regions of Spain and OXA-48-like in the rest of the country. Regarding P. aeruginosa, VIM is widely distributed all over the country. Globally, an increasing percentage of OXA-48-like enzymes was observed from 2014 to 2017. KPC enzymes were more frequent in 2017-2018 compared to 2014-2016. Discussion: Data from this study help to understand the situation and evolution of the main species of CPE and CPPA in Spain, with practical implications for control and optimal treatment of infections caused by these multi-drug resistant organisms.

Comparación entre cinco técnicas de PCR para el diagnóstico del SARS-CoV-2 / Comparison between five PCR techniques for the diagnosis of SARS-CoV-2

Introducción. Desde que aparecieron los primeros casosde SARS-CoV-2 son numerosas las técnicas que se han desarrollado para el diagnóstico o seguimiento de la infección,tanto técnicas directas como serológicas. La elección de unabuena herramienta diagnóstica es fundamental para el controlepidemiológico. El objetivo ha sido comparar cinco técnicascomercializadas de RT-PCR a tiempo real, en sensibilidad, especificidad y concordancia para la detección del SARS-CoV-2.Material y métodos. Se compararon cinco kits comerciales de RT-PCR para la detección del SARS-CoV-2. Se tomaronocho muestras positivas conocidas que se sometieron a sietediluciones o concentraciones diferentes y otras 135 muestrasnegativas para determinar valores de sensibilidad, especificidad y concordancia.Resultados. La sensibilidad, especificidad, valor predictivopositivo (VPP) y valor predictivo negativo (VPN) para las técnicas de Palex, Roche y GeneXpert respecto a Seegene fueronidénticas, correspondientes a 98,21%, 100%, 100% y 99,26%respectivamente. Para Becton Dickinson la sensibilidad fue del89,28%, la especificidad del 100%, el VPP del 100% y el VPNdel 95,74%. La concordancia mediante el índice Kappa paraPalex, Roche y GeneXpert fue del 0,9892, mientras que la concordancia para Becton Dickinson fue con un índice Kappa de0,9215.Conclusión. Todos los kits de RT-PCR comerciales presentaron elevadas sensibilidades y especificidades, así como VPP,VPN y concordancia. (AU)

Introduction. Since the first cases of SARS-CoV-2appeared, there have been numerous techniques that havebeen developed for the diagnosis or monitoring of infection, both direct and serological techniques. Choosing agood diagnostic tool is essential for epidemiological control. The objective was to compare five commercializedRT-PCR techniques in real time, in sensitivity, specificityand agreement for the detection of SARS-CoV-2.Material and methods. Five commercial RT-PCR kitsfor the detection of SARS-CoV-2 were compared. Eightknown positive samples were taken and subjected to seven different dilutions or concentrations, and another 135negative samples were used to determine sensitivity, specificity, and agreement values.Results. The sensitivity, specificity, positive predictivevalue (PPV) and negative predictive value (NPV) for thePalex, Roche and GeneXpert techniques with respect toSeegene were identical, corresponding to 98.21%, 100%,100% and 99.26% respectively. For Becton Dickinson thesensitivity was 89.28%, the specificity of 100%, the PPVof 100% and the NPV of 95.74%. The agreement using theKappa index for Palex, Roche and GeneXpert was 0.9892,while the agreement for Becton Dickinson was with aKappa index of 0.9215.Conclusion. All commercial RT-PCR kits had highsensitivities and specificities, as well as PPV, NPV, andconcordance. (AU)

Antibody response in patients admitted to the hospital with suspected SARS-CoV-2 infection: results from a multicenter study across Spain.

AIM: To evaluate the serological response against SARS-CoV-2 in a multicenter study representative of the Spanish COVID pandemic. METHODS: IgG and IgM + IgA responses were measured on 1466 samples from 1236 Spanish COVID-19 patients admitted to the hospital, two commercial ELISA kits (Vircell SL, Spain) based on the detection of antibodies against the viral spike protein and nucleoprotein, were used. RESULTS: Approximately half of the patients presented antibodies (56.8% were IgM + IgA positive and 43.0% were IgG positive) as soon as 2 days after the first positive PCR result. Serological test positivity increased with time from the PCR test, and 10 days after the first PCR result, 91.5% and 88.0% of the patients presented IgM + IgA and IgG antibodies, respectively. CONCLUSION: The high values of sensitivity attained in the present study from a relatively early period of time after hospitalization support the use of the evaluated serological assays as supplementary diagnostic tests for the clinical management of COVID-19.

Diagnóstico microbiológico de las infecciones asociadas a dispositivos biomédicos / Microbiological diagnosis of medical device-associated infections

El empleo de dispositivos biomédicos implantados quirúrgicamente se ha incrementado en los últimos años. A pesar de las mejoras en las técnicas quirúrgicas, en los materiales y el diseño de los dispositivos, la infección asociada continúa siendo una complicación relativamente frecuente y grave. La infección se produce generalmente durante la cirugía a partir de la microbiota cutánea del paciente. Cuando los microorganismos colonizan el dispositivo se desarrollan sobre su superficie formando una biocapa que es determinante en la patogenia de estas infecciones. El diagnóstico microbiológico es difícil y en muchas ocasiones solo se consigue tras la retirada del dispositivo. El cultivo tras sonicación puede ser una herramienta diagnóstica útil ya que consigue la desagregación de la biocapa. También, las técnicas moleculares, especialmente las basadas en PCR, aplicadas a tejidos y al material obtenido tras sonicación han demostrado alta sensibilidad y especificidad en el diagnóstico de infecciones asociadas a dispositivos intracardiacos

The use of surgically implanted medical devices has increased greatly over the last few years. Despite surgical advances and improvements in the materials and design of devices, infection continues to be a major complication of their use. Device-associated infections are produced mainly during their implantation and, are caused by microorganisms that are part of the skin flora. Biofilm development on device surfaces is the most important factor to explain the pathophysiological aspects of infection. Microbiological diagnosis is difficult and can often only be achieved after removal of the device. Sonication of the removed device may be a useful tool, since this procedure dislodges and disaggregates biofilm bacteria from the device. Molecular techniques, especially PCR, applied to the tissues and material obtained after sonication have shown to have a high sensitivity and specificity for the diagnosis of cardiovascular device infections

[Microbiological diagnosis of medical device-associated infections]. / Diagnóstico microbiológico de las infecciones asociadas a dispositivos biomédicos.

The use of surgically implanted medical devices has increased greatly over the last few years. Despite surgical advances and improvements in the materials and design of devices, infection continues to be a major complication of their use. Device-associated infections are produced mainly during their implantation and, are caused by microorganisms that are part of the skin flora. Biofilm development on device surfaces is the most important factor to explain the pathophysiological aspects of infection. Microbiological diagnosis is difficult and can often only be achieved after removal of the device. Sonication of the removed device may be a useful tool, since this procedure dislodges and disaggregates biofilm bacteria from the device. Molecular techniques, especially PCR, applied to the tissues and material obtained after sonication have shown to have a high sensitivity and specificity for the diagnosis of cardiovascular device infections.

Paciente con blefaritis crónica resistente a los tratamientos oftalmológicos habituales / Resistance to routine ophthalmological treatments in a patient with chronic blepharitis

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Factores de riesgo de neumonía nosocomial por Staphylococcus aureus resistente a meticilina / Risk factors of nosocomial pneumonia caused by methicillin-resistant Staphylococcus aureus

Fundamento y objetivo: Es necesario predecir una neumonía nosocomial (NN) por Staphylococcus aureus resistente a meticilina (SARM) para facilitar la inclusión de un antibiótico específico en la terapia empírica. En este estudio se desarrolla un modelo para predecir la probabilidad de NN por SARM cuando se desconoce el estado de portador y el diagnóstico microbiológico. Pacientes y método: Se diseñó un estudio de casos y controles, realizándose una regresión logística multivariable para identificar los factores de riesgo de NN por SARM. Se incluyeron factores demográficos, relacionados con la hospitalización, la inmunodepresión, neutropenia, la medicación y la gravedad.Resultados: Se estudiaron 363 pacientes (121 casos y 242 controles). Permanecieron en el modelo final la edad>14 años (odds ratio [OR] 7,4, intervalo de confianza del 95% [IC 95%] 1,5-37,4, p<0,015), la aparición de la NN>6 días después del ingreso (OR 4,1, IC 95% 2,4-7,1, p<0,001), el desarrollo de la NN fuera del verano (OR 2,5, IC 95% 1,2-5,2, p = 0,015), las enfermedades respiratorias (OR 4,9, IC 95% 1,5-15,8, p = 0,007) y la afectación multilobar (OR 4, IC 95% 2,3-7,2, p<0,001). Con estas variables se calculó la probabilidad de desarrollar neumonía por SARM para cada una de las posibles combinaciones, clasificándose en criterios mayores y menores.Conclusiones: Se debe incluir cobertura de SARM en el tratamiento empírico de la NN cuando: a) un paciente adulto (>14 años) tiene, al menos, 2 criterios mayores o un criterio mayor y 2 menores, y b) un paciente<14 años tiene los 2 criterios mayores y los 2 menores (AU)

Background and objective: To include a specific antibiotic in the empiric therapy, it is necessary to predict when a nosocomial pneumonia (NP) is caused by methicillin-resistant Staphylococcus aureus (MRSA). We have developed a model for the prediction of the probability of a NP being caused by MRSA, when the carrier status and the microbiological diagnosis are unknown. Patients and methods: A retrospective case-control study (1999-2005) was designed. A univariate and multivariate logistic regression was performed to identify the risk factors for suffering a NP due to MRSA. Demographic factors, related to hospitalization, immunosuppression or neutropenia, to medication and severity were included. Results:Three hundred and sixty three patients (121 cases and 242 controls) were studied. The final model of multivariate logistic regression included an age>14 years (OR 7.4, CI 95% 1.5-37.4, P<.015), NP appearance>6 days after admittance (OR 4.1, CI 95% 2.4-7,1, P<.001), NP development excluding summers (OR 2.5, CI 95% 1.2-5.2, P<.015), respiratory diseases (OR 4.9, CI 95% 1.5-15.8, P<.007) and multilobar involvement (OR 4, CI 95% 2.3-7.2, P<.001).The probability of developing a pneumonia due to MRSA was studied for each of the possible combinations and subsequently classified in minor and major criteria. Conclusions: MRSA coverage should be included in the empirical treatment of NP when: a) an adult patient (>14 years old) presents, at least, 2 major criteria or 1 major criterion together with 2 minor criteria, and b) a patient <14 years-old has 2 major criteria as well as 2 minor criteria Ç(AU)

[Risk factors of nosocomial pneumonia caused by methicillin-resistant Staphylococcus aureus]. / Factores de riesgo de neumonía nosocomial por Staphylococcus aureus resistente a meticilina.

BACKGROUND AND OBJECTIVE: To include a specific antibiotic in the empiric therapy, it is necessary to predict when a nosocomial pneumonia (NP) is caused by methicillin-resistant Staphylococcus aureus (MRSA). We have developed a model for the prediction of the probability of a NP being caused by MRSA, when the carrier status and the microbiological diagnosis are unknown. PATIENTS AND METHODS: A retrospective case-control study (1999-2005) was designed. A univariate and multivariate logistic regression was performed to identify the risk factors for suffering a NP due to MRSA. Demographic factors, related to hospitalization, immunosuppression or neutropenia, to medication and severity were included. RESULTS: Three hundred and sixty three patients (121 cases and 242 controls) were studied. The final model of multivariate logistic regression included an age>14 years (OR 7.4, CI 95% 1.5-37.4, P<.015), NP appearance>6 days after admittance (OR 4.1, CI 95% 2.4-7,1, P<.001), NP development excluding summers (OR 2.5, CI 95% 1.2-5.2, P<.015), respiratory diseases (OR 4.9, CI 95% 1.5-15.8, P<.007) and multilobar involvement (OR 4, CI 95% 2.3-7.2, P<.001).The probability of developing a pneumonia due to MRSA was studied for each of the possible combinations and subsequently classified in minor and major criteria. CONCLUSIONS: MRSA coverage should be included in the empirical treatment of NP when: a) an adult patient (>14 years old) presents, at least, 2 major criteria or 1 major criterion together with 2 minor criteria, and b) a patient <14 years-old has 2 major criteria as well as 2 minor criteria.

Paciente procedente de la República del Congo con dolor ocular intermitente / Patient from the Republic of Congo with intermittent ocular pain

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