Derivation and external validation of a simple prediction rule for the development of respiratory failure in hospitalized patients with influenza.

BACKGROUND: Influenza viruses cause seasonal epidemics worldwide with a significant morbimortality burden. Clinical spectrum of Influenza is wide, being respiratory failure (RF) one of its most severe complications. This study aims to elaborate a clinical prediction rule of RF in hospitalized Influenza patients. METHODS: A prospective cohort study was conducted during two consecutive Influenza seasons (December 2016-March 2017 and December 2017-April 2018) including hospitalized adults with confirmed A or B Influenza infection. A prediction rule was derived using logistic regression and recursive partitioning, followed by internal cross-validation. External validation was performed on a retrospective cohort in a different hospital between December 2018 and May 2019. RESULTS: Overall, 707 patients were included in the derivation cohort and 285 in the validation cohort. RF rate was 6.8% and 11.6%, respectively. Chronic obstructive pulmonary disease, immunosuppression, radiological abnormalities, respiratory rate, lymphopenia, lactate dehydrogenase and C-reactive protein at admission were associated with RF. A four category-grouped seven point-score was derived including radiological abnormalities, lymphopenia, respiratory rate and lactate dehydrogenase. Final model area under the curve was 0.796 (0.714-0.877) in the derivation cohort and 0.773 (0.687-0.859) in the validation cohort (p < 0.001 in both cases). The predicted model showed an adequate fit with the observed results (Fisher's test p > 0.43). CONCLUSION: we present a simple, discriminating, well-calibrated rule for an early prediction of the development of RF in hospitalized Influenza patients, with proper performance in an external validation cohort. This tool can be helpful in patient's stratification during seasonal Influenza epidemics.

Implementación generalizada del diagnóstico en un solo paso (DUSP) de la hepatitis C en Galicia: resultados para la reflexión / Generalized implementation of reflex testing of hepatitis C in Galicia: Results for reflection

Introducción: La implementación del diagnóstico en un solo paso (DUSP) de la infección activa por virus de la hepatitis C (VHC) conjuntamente con la incorporación de alertas informativas ha demostrado que reduce de forma significativa, respecto al diagnóstico tradicional, la cifra de pacientes que no eran remitidos para valoración terapéutica. Métodos: A partir de la implementación en 2018 del DUSP en los servicios de microbiología de los hospitales del Servicio Gallego de Salud (SERGAS), se identifican y caracterizan de manera retrospectiva los nuevos diagnósticos de infección activa por VHC. Resultados: En 2018 se identificaron mediante DUSP un total de 258 pacientes con infección activa por VHC desconocida (70,2% hombres, mediana de edad de 52 años) procedentes de consultas de unidades de atención primaria y especializada en un 54,8% y 39,8%, respectivamente, así como de otras localizaciones en un 5,4%. De los 258 pacientes, el 81,0% fueron derivados para valoración terapéutica, con una mediana de 54 días desde su diagnóstico. En el 58,3% de los casos se determinó el DUSP mediante carga viral, el genotipo predominante fue el 1a (30,7%), un 52,1% fue tratado y se observó una respuesta viral sostenida en el 93,7% de estos. Conclusión: La implementación en toda Galicia del DUSP de la hepatitis C conjuntamente con alertas informativas ha permitido obtener, en conjunto, tasas de derivación para tratamiento similares a las obtenidas en otros estudios. Sin embargo, existe una amplia variabilidad entre los distintos centros, que exigen la incorporación de mejoras, como la formación o la utilización de medidas de rescate para su optimización.(AU)

Introduction: The implementation of reflex testing of active hepatitis C virus (HCV) infection, together with the incorporation of informative alerts in the reports, has shown that it significantly reduces the number of patients who were not referred for therapeutic evaluation. Methods: Since the implementation in 2018 of the DUSP in the microbiology services of the Galician Health Service hospitals (SERGAS), new diagnoses of active HCV infection have been retrospectively identified and characterized. Results: In 2018, a total of 258 patients with unknown active HCV infection (70,2% men, middle age 52 years) were identified through by reflex testing from consultations of primary and specialized care units in 54.8% and 39.8% respectively, as well as from other locations by 5.4%. Of the 258 patients, 81.0% were referred for therapeutic evaluation, with a median of 54 days from their diagnosis. In 58.3% of the cases the reflex testing was determined by viral load, the predominant genotype was 1a (30,7%) and 52,1% were treated, observing sustained viral response in 93.7% of these. Conclusion: The generalized implementation of the HCV reflex testing together with informative alerts in Galicia has allowed us to obtain referral rates for treatment similar to those obtained in other studies. However, there is a wide variability between the different centers that require the incorporation of improvements, such as training or the use of rescue measures for optimization.(AU)

Viruses Previously Identified in Brazil as Belonging to HIV-1 CRF72_BF1 Represent Two Closely Related Circulating Recombinant Forms, One of Which, Designated CRF122_BF1, Is Also Circulating in Spain.

Circulating recombinant forms (CRFs) are important components of the HIV-1 pandemic. Those derived from recombination between subtype B and subsubtype F1, with 18 reported, most of them of South American origin, are among the most diverse. In this study, we identified a HIV-1 BF1 recombinant cluster that is expanding in Spain, transmitted mainly via heterosexual contact, which, analyzed in near full-length genomes in four viruses, exhibited a coincident BF1 mosaic structure, with 12 breakpoints, that fully coincided with that of two viruses (10BR_MG003 and 10BR_MG005) from Brazil, previously classified as CRF72_BF1. The three remaining Brazilian viruses (10BR_MG002, 10BR_MG004, and 10BR_MG008) previously identified as CRF72_BF1 exhibited mosaic structures highly similar, but not identical, to that of the Spanish viruses and to 10BR_MG003 and 10BR_MG005, with discrepant subtypes in two short genome segments, located in pol and gp120env. Based on these results, we propose that the five viruses from Brazil previously identified as CRF72_BF1 actually belong to two closely related CRFs, one comprising 10BR_MG002, 10BR_MG004, and 10BR_MG008, which keep their CRF72_BF1 designation, and the other, designated CRF122_BF1, comprising 10BR_MG003, 10BR_MG005, and the viruses of the identified Spanish cluster. Three other BF1 recombinant genomes, two from Brazil and one from Italy, previously identified as unique recombinant forms, were classified as CRF72_BF1. CRF122_BF1, but not CRF72_BF1, was associated with protease L89M substitution, which was reported to contribute to antiretroviral drug resistance. Phylodynamic analyses estimate the emergence of CRF122_BF1 in Brazil around 1987. Given their close phylogenetic relationship and similar structures, the grouping of CRF72_BF1 and CRF122_BF1 in a CRF family is proposed.

Generalized implementation of reflex testing of hepatitis C in Galicia: Results for reflection.

INTRODUCTION: The implementation of reflex testing of active hepatitis C virus (HCV) infection, together with the incorporation of informative alerts in the reports, has shown that it significantly reduces the number of patients who were not referred for therapeutic evaluation. METHODS: Since the implementation in 2018 of the DUSP in the Microbiology Services of the Galician Health Service hospitals (SERGAS), new diagnoses of active HCV infection have been retrospectively identified and characterized. RESULTS: In 2018, a total of 258 patients with unknown active HCV infection (70,2% men, middle age 52 years) were identified through by reflex testing from consultations of primary and specialized care units in 54.8% and 39.8% respectively, as well as from other locations by 5.4%. Of the 258 patients, 81.0% were referred for therapeutic evaluation, with a median of 54 days from their diagnosis. In 58.3% of the cases the reflex testing was determined by viral load, the predominant genotype was 1a (30,7%) and 52,1% were treated, observing sustained viral response (SVR) in 93.7 % of these. CONCLUSION: The generalized implementation of the HCV reflex testing together with informative alerts in Galicia has allowed us to obtain referral rates for treatment similar to those obtained in other studies. However, there is a wide variability between the different centers that require the incorporation of improvements, such as training or the use of rescue measures for optimization.

Generalized implementation of reflex testing of hepatitis C in Galicia: Results for reflection. / Implementación generalizada del diagnóstico en un solo paso (DUSP) de la hepatitis C en Galicia: resultados para la reflexión.

INTRODUCTION: The implementation of reflex testing of active hepatitis C virus (HCV) infection, together with the incorporation of informative alerts in the reports, has shown that it significantly reduces the number of patients who were not referred for therapeutic evaluation. METHODS: Since the implementation in 2018 of the DUSP in the microbiology services of the Galician Health Service hospitals (SERGAS), new diagnoses of active HCV infection have been retrospectively identified and characterized. RESULTS: In 2018, a total of 258 patients with unknown active HCV infection (70,2% men, middle age 52 years) were identified through by reflex testing from consultations of primary and specialized care units in 54.8% and 39.8% respectively, as well as from other locations by 5.4%. Of the 258 patients, 81.0% were referred for therapeutic evaluation, with a median of 54 days from their diagnosis. In 58.3% of the cases the reflex testing was determined by viral load, the predominant genotype was 1a (30,7%) and 52,1% were treated, observing sustained viral response in 93.7% of these. CONCLUSION: The generalized implementation of the HCV reflex testing together with informative alerts in Galicia has allowed us to obtain referral rates for treatment similar to those obtained in other studies. However, there is a wide variability between the different centers that require the incorporation of improvements, such as training or the use of rescue measures for optimization.

Sample pooling for SARS-CoV-2 RT-PCR screening.

OBJECTIVE: To evaluate the efficacy of sample pooling compared to the individual analysis for the diagnosis of coronavirus disease 2019 (COVID-19) by using different commercial platforms for nucleic acid extraction and amplification. METHODS: A total of 3519 nasopharyngeal samples received at nine Spanish clinical microbiology laboratories were processed individually and in pools (342 pools of ten samples and 11 pools of nine samples) according to the existing methodology in place at each centre. RESULTS: We found that 253 pools (2519 samples) were negative and 99 pools (990 samples) were positive; with 241 positive samples (6.85%), our pooling strategy would have saved 2167 PCR tests. For 29 pools (made out of 290 samples), we found discordant results when compared to their correspondent individual samples, as follows: in 22 of 29 pools (28 samples), minor discordances were found; for seven pools (7 samples), we found major discordances. Sensitivity, specificity and positive and negative predictive values for pooling were 97.10% (95% confidence interval (CI), 94.11-98.82), 100%, 100% and 99.79% (95% CI, 99.56-99.90) respectively; accuracy was 99.80% (95% CI, 99.59-99.92), and the kappa concordant coefficient was 0.984. The dilution of samples in our pooling strategy resulted in a median loss of 2.87 (95% CI, 2.46-3.28) cycle threshold (Ct) for E gene, 3.36 (95% CI, 2.89-3.85) Ct for the RdRP gene and 2.99 (95% CI, 2.56-3.43) Ct for the N gene. CONCLUSIONS: We found a high efficiency of pooling strategies for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA testing across different RNA extraction and amplification platforms, with excellent performance in terms of sensitivity, specificity and positive and negative predictive values.

High incidence of MDR and XDR Pseudomonas aeruginosa isolates obtained from patients with ventilator-associated pneumonia in Greece, Italy and Spain as part of the MagicBullet clinical trial.

OBJECTIVES: To characterize the antimicrobial susceptibility, molecular epidemiology and carbapenem resistance mechanisms in Pseudomonas aeruginosa isolates recovered from respiratory tract samples from patients with ventilator-associated pneumonia enrolled in the MagicBullet clinical trial. METHODS: Isolates were collected from 53 patients from 12 hospitals in Spain, Italy and Greece. Susceptibility was determined using broth microdilution and Etest. MALDI-TOF MS was used to detect carbapenemase activity and carbapenemases were identified by PCR and sequencing. Molecular epidemiology was investigated using PFGE and MLST. RESULTS: Of the 53 isolates, 2 (3.8%) were considered pandrug resistant (PDR), 19 (35.8%) were XDR and 16 (30.2%) were MDR. Most (88.9%) of the isolates from Greece were MDR, XDR or PDR, whereas fewer of the isolates from Spain (33.3%) and Italy (43.5%) showed antibiotic resistance. Three Greek isolates were resistant to colistin. Overall, the rates of resistance of P. aeruginosa isolates to imipenem, ciprofloxacin, ceftolozane/tazobactam and ceftazidime/avibactam were 64.1%, 54.7%, 22.6% and 24.5%, respectively. All isolates resistant to ceftolozane/tazobactam and ceftazidime/avibactam (Greece, n = 10; and Italy, n = 2) carried blaVIM-2. Spanish isolates were susceptible to the new drug combinations. Forty-eight restriction patterns and 27 STs were documented. Sixty percent of isolates belonged to six STs, including the high-risk clones ST-111, ST-175 and ST-235. CONCLUSIONS: MDR/XDR isolates were highly prevalent, particularly in Greece. The most effective antibiotic against P. aeruginosa was colistin, followed by ceftolozane/tazobactam and ceftazidime/avibactam. blaVIM-2 is associated with resistance to ceftolozane/tazobactam and ceftazidime/avibactam, and related to highly resistant phenotypes. ST-111 was the most frequent and disseminated clone and the clonal diversity was lower in XDR and PDR strains.

Use of the cobas 4800 system for the rapid detection of toxigenic Clostridium difficile and methicillin-resistant Staphylococcus aureus.

The new cobas® Cdiff and cobas® MRSA/SA tests were compared with conventional methods for the rapid detection of toxigenic Clostridium difficile and methicillin-resistant Staphylococcus aureus. The final concordance between cobas Cdiff Test and GDH/toxin gene screening was 97.62% and between cobas MRSA/SA Test and chromogenic culture, 91.30%, respectively.

Characterization of plasmid-mediated ß-lactamases in fecal colonizing patients in the hospital and community setting in Spain.

AIM: Active surveillance of plasmid-mediated ß-lactamase-producing Enterobacteriaceae (PMBL-E) in fecal carriers in the hospital and in the community setting in a non-outbreak period of time. METHODS: Patients were screened for carriage of Enterobacteriaceae resistant to expanded-spectrum cephalosporins and PMBL-E were characterized (extended-spectrum-ß-lactamase [ESBL], plasmid-mediated AmpC ß-lactamase [pAmpC], and carbapenemases) by PCR and sequencing. RESULTS: The prevalence of ESBL and pAmpC carriers was 5.06% and 0.59%, respectively. Overall, CTX-M-like enzymes were the ESBL dominate enzymes (96.15%). The group CTX-M-9 was the most prevalent (81, 54%) [CTX-M-14 (74, 91.35%), CTX-M-9 (5, 6.17%), CTX-M-24 (1, 1.23%), and CTX-M-27 (1, 1.23%)] followed by the group CTX-M-1 (64, 42.67%) [CTX-M-15 (42, 65.63%), CTX-M-1 (13, 20.31%), CTX-M-32 (8, 12.5%), and CTX-M-3 (1, 1.56%)]. One CTX-M-10, one CTX-M-59, and three CTX-M-8 were also found. A very small representation of SHV or TEM ESBL enzymes was found (3.2% and 0.64%, respectively). pAmpC characterization revealed a predominance of CMY-2 (81.25%), followed by DHA-1 (18.75%). We did not detect the presence of carbapenemase producers. CONCLUSIONS: The prevalence of ESBL-producers from fecal carriers is stable in our area, but colonization by pAmpC producers has emerged recently as we have confirmed. Periodic active surveillance is useful to identify these human reservoirs and control the evolution of PMBL carriage in a community over time.

Epidemiological features, resistance genes, and clones among community-onset methicillin-resistant Staphylococcus aureus (CO-MRSA) isolates detected in northern Spain.

Twenty-nine community-onset methicillin-resistant Staphylococcus aureus (CO-MRSA) isolates were prospectively selected according to epidemiological criteria among 374 MRSA isolates collected in our laboratory during 2009-2010 in order to determine which community-associated MRSA (CA-MRSA) and healthcare-associated MRSA (HA-MRSA) clones are circulating in the community in northern Spain. PVL genes were detected in 5 strains (17.2%) that belonged to SCCmec type IV or V and to the agr group I (ST8 and ST2050), agr group II (ST121), and agr group III (ST30 and ST852). These strains were isolated from patients with different clinical manifestations such as urinary tract infection, abscess, or pneumonia, and most of them belonged to emergency department patients with no history of visits to General Practitioners (GPs) in the year before the isolation. We considered that the prevalence of CA-MRSA in community-onset isolates was low (17.2%). A high proportion of the CO-MRSA strains (58.6%) were ST125-MRSA-IVc (CC5), responsible for most of the infections caused by HA-MRSA strains in Spain. This endemic clone is also circulating in the community of northern Spain as we could demonstrate in this study. Antimicrobial resistance was found in spa type t067 isolates linked to the presence of ant(4')-Ia and msr(A). Most of the CO-MRSA isolates in this study corresponded to spa types more associated to the hospital environment, suggesting the interchange of genetic lineages of MRSA among community and hospital niches.

Caracterización de Shigella sonnei portadora de CTX-M-15 en un paciente español sin antecedentes de viaje al extranjero / Characterisation of a CTX-M-15-producing Shigella sonnei in a Spanish patient who had not travelled abroad

De las 107 Shigella spp. aisladas entre 2000-2010, en una cepa de Shigella sonnei se detectaron los genes codificantes de TEM-1 y CTX-M-15, identificándose la estructura ISEcp1+blaCTX-M-15+orf477. La cepa portaba un integrón de clase 2 con los genes cassettes dfrA1+sat+aadA1. Se detectó un plásmido del grupo IncI1 ST31 (CC-31) demostrando su transferencia por conjugación. Describimos la primera cepa de S. sonnei en nuestra comunidad productora de CTX-M-15 perteneciente a un paciente español que no había viajado al extranjero (AU)

One hundred and seven Shigella spp. strains were isolated in our laboratory during the years 2000 to2010. One Shigella sonnei harboured the genes that coded the -lactamases TEM-1 and CTX-M-15, identifying the structure, ISEcp1 + blaCTX-M-15 + orf477, in their genetic environment. The strain also carrieda class 2 integron with the gene cassettes dfrA1 + sat + aadA1. A plasmid group IncI1 ST31 (CC-31)was detected and its mobilization by conjugation was demonstrated. We describe for the (..) (AU)

[Emergence of plasmid mediated AmpC ß-lactamasas: Origin, importance, detection and therapeutical options]. / Emergencia de ß-lactamasas AmpC plasmídicas (pAmpC ó cefamicinasas): origen, importancia, detección y alternativas terapéuticas.

AmpC ß-lactamases can hydrolyze penicillins, oxyimino-, 7-α-methoxycephalosporins and monobactams. Susceptibility to cefepime or cefpirome is little affected and is unchanged for carbapenems. Originally such genes are thought to have been mobilized to mobile genetic elements from the chromosomal ampC genes from members of Enterobacteriaceae facilitating their spread and now they can appear in bacterial lacking or poorly expressing a chromosomal ampC gene. The prevalence of infection by plasmid mediated AmpC (pAmpC) varies depending on the type of enzyme and geographical location and blaCMY-2 is the most frequently detected worldwide. Typically, pAmpC producing isolates are associated with resistance to multiple antibiotics making the selection of an effective antibiotic difficult. Phenotypic and molecular methods to detect pAmpC are described and the role of different antibiotics in the treatment of these infections is examined. Surveillance studies about the evolution of this emerging resistant mechanism are important in clinical isolates. Evaluate the in vitro susceptibility of these isolates and the clinical efficacy of other therapeutic options is required.

[Characterisation of a CTX-M-15-producing Shigella sonnei in a Spanish patient who had not travelled abroad]. / Caracterización de Shigella sonnei portadora de CTX-M-15 en un paciente español sin antecedentes de viaje al extranjero.

One hundred and seven Shigella spp. strains were isolated in our laboratory during the years 2000 to 2010. One Shigella sonnei harboured the genes that coded the ß-lactamases TEM-1 and CTX-M-15, identifying the structure, ISEcp1+bla(CTX-M-15)+orf477, in their genetic environment. The strain also carried a class 2 integron with the gene cassettes dfrA1+sat+aadA1. A plasmid group IncI1 ST31 (CC-31) was detected and its mobilization by conjugation was demonstrated. We describe for the first time a S. sonnei strain producing a CTX-M-15 ß-lactamase recovered from a Spanish patient who had not travelled abroad.