Diagnostic tool for surveillance, detection and monitoring of the high-risk clone K. pneumoniae ST15.

BACKGROUND: The global spread of Klebsiella pneumoniae ST15, causing multi-continental outbreaks, contributes to the movement of resistance genes between clones increasing the antimicrobial resistance crisis. The genomic traits providing it with the ability to outcompete other bacteria and cause epidemics remain unclear. AIM: To identify the specific genomic traits of K. pneumoniae ST15 to develop a diagnostic test. METHODS: An outbreak caused by K. pneumoniae occurred in Hospital A Coruña, Spain. Antimicrobial susceptibility analysis and molecular typing (PGFE and MLST) were performed. One isolate of each sequence type was selected for whole-genome sequencing analysis. Comparative analysis of genomes was performed using RAST. BLASTn was used to evaluate the presence of the fhaC and kpiD genes. Two hundred and ninety-four K. pneumoniae from a Spanish nationwide collection were analysed by PCR. FINDINGS: Genotyping showed that 87.5% of the isolates tested belonged to a clone with a unique PFGE pattern which corresponded to ST15. Comparative genomic analysis of the different STs enabled us to determine the specific genomic traits of K. pneumoniae ST15. Two adherence-related systems (Kpi and KpFhaB/FhaC) were specific markers of this clone. Multiplex-PCR analysis with kpiD and fhaC oligonucleotides revealed that K. pneumoniae ST15 is specifically detected with a sensitivity of 100% and a specificity of 97.76%. The PCR results showed 100% concordance with the MLST and whole-genome sequencing data. CONCLUSION: K. pneumoniae ST15 possesses specific genomic traits that could favour its dissemination. They could be used as targets to detect K. pneumoniae ST15 with high sensitivity and specificity.

Draft Genome Sequences of Two Epidemic OXA-48-Producing Klebsiella pneumoniae Clinical Strains Isolated during a Large Outbreak in Spain.

We report here the draft genome sequences of Klebsiella pneumoniae strains Kp1803 and Kp3380 isolated during a large outbreak at A Coruña Hospital in Spain. The final genome assemblies for Kp1803 and Kp3380 comprise approximately 6.6 and 6.1 Mb, respectively, and both strains have G+C contents of 57.2%.

Genomic Evolution of Two Acinetobacter baumannii Clinical Strains from ST-2 Clones Isolated in 2000 and 2010 (ST-2_clon_2000 and ST-2_clon_2010).

Acinetobacter baumannii is a successful nosocomial pathogen due to its ability to persist in hospital environments by acquiring mobile elements such as transposons, plasmids, and phages. In this study, we compared two genomes of A. baumannii clinical strains isolated in 2000 (ST-2_clon_2000) and 2010 (ST-2_clon_2010) from GenBank project PRJNA308422.

Genome Sequence of a Clinical Strain of Acinetobacter baumannii Belonging to the ST79/PFGE-HUI-1 Clone Lacking the AdeABC (Resistance-Nodulation-Cell Division-Type) Efflux Pump.

Increased expression of chromosomal genes for resistance-nodulation-cell division-type efflux systems plays a major role in the multidrug resistance of Acinetobacter baumannii Little is known about the genetic characteristics of clinical strains of Acinetobacter baumannii lacking the AdeABC pump. In this study, we sequenced the genome of clinical strain Ab421 GEIH-2010 (belonging to clone ST79/PFGE-HUI-1 from the GEIH-REIPI Ab. 2010 project) which lacks this efflux pump.

Contribution of efflux pumps, porins, and ß-lactamases to multidrug resistance in clinical isolates of Acinetobacter baumannii.

We investigated the mechanisms of resistance to carbapenems, aminoglycosides, glycylcyclines, tetracyclines, and quinolones in 90 multiresistant clinical strains of Acinetobacter baumannii isolated from two genetically unrelated A. baumannii clones: clone PFGE-ROC-1 (53 strains producing the OXA-58 ß-lactamase enzyme and 18 strains with the OXA-24 ß-lactamase) and clone PFGE-HUI-1 (19 strains susceptible to carbapenems). We used real-time reverse transcriptase PCR to correlate antimicrobial resistance (MICs) with expression of genes encoding chromosomal ß-lactamases (AmpC and OXA-51), porins (OmpA, CarO, Omp33, Dcap-like, OprB, Omp25, OprC, OprD, and OmpW), and proteins integral to six efflux systems (AdeABC, AdeIJK, AdeFGH, CraA, AbeM, and AmvA). Overexpression of the AdeABC system (level of expression relative to that by A. baumannii ATCC 17978, 30- to 45-fold) was significantly associated with resistance to tigecycline, minocycline, and gentamicin and other biological functions. However, hyperexpression of the AdeIJK efflux pump (level of expression relative to that by A. baumannii ATCC 17978, 8- to 10-fold) was significantly associated only with resistance to tigecycline and minocycline (to which the TetB efflux system also contributed). TetB and TetA(39) efflux pumps were detected in clinical strains and were associated with resistance to tetracyclines and doxycycline. The absence of the AdeABC system and the lack of expression of other mechanisms suggest that tigecycline-resistant strains of the PFGE-HUI-1 clone may be associated with a novel resistance-nodulation-cell efflux pump (decreased MICs in the presence of the inhibitor Phe-Arg ß-naphthylamide dihydrochloride) and the TetA(39) system.

First report of an OXA-23 carbapenemase-producing Acinetobacter baumannii clinical isolate related to Tn2006 in Spain.

A carbapenem-resistant Acinetobacter baumannii clinical isolate belonging to European clone II and sequence type 2 was recovered from a patient in the Son Espases hospital in Mallorca, Spain. Genetic analysis showed the presence of the bla(OXA-23) gene in association with the widely disseminated transposon Tn2006. This is the first reported identification of A. baumannii carrying bla(OXA-23) in Spain.

Estudio observacional, prospectivo y multicéntrico del tratamiento ambulatorio de la trombosis venosa en fase aguda en el sector femoropopliteotibial / A prospective observational and multicentre study of the treatment of acute-phase venous thrombosis in the femoral -popliteal-tibial region in outpatients

Introducción. El tratamiento de la trombosis venosa (TVP) mediante heparina fraccionada de bajo peso molecular (HBPM) ha facultado que diversos grupos hayan iniciado su tratamiento deforma extrahospitala-ria. Objetivos. Evaluar la eficacia y seguridad del tratamiento extrahospitalario de la TVP en fase aguda y la efectividad y tolerabilidad de la HBPM (tinzaparina). Pacientes y métodos. Estudio observacional, prospectivo y multicéntrico (15 hospitales) sobre 122 pacientes: 57, 4 por ciento mujeres y 42,6 por ciento varones, con una edad media de 60,9 años. Los criterios de inclusión fueron: TVP en el sector femoropopliteotibial, y los de exclusión: TVP primaria o recidivante en el sector iliocavo, edad superior a 75 años, patología sistémica grave y alergia documentada a la tinzaparina. Los criterios de valoración de la eficacia y la seguridad fueron: intercurrencia de complicaciones (embolismo pulmonar, episodios de sangrado), progresión de la trombosis, parámetros clínicos (edema) y de calidad de vida SF-12 (escala EVA). Para la evaluación de las reacciones adversas se empleó la clasificación WHOART. El período medio de tratamiento fue de 11,6 días. Resultados. a) Eficacia: intercurrencia trombótica, 0 por ciento; embolismo pulmonar no fatal: 1(0,82 por ciento; ingresos hospitalarios: 4 (3,2 por ciento); perímetro de la extremidad: regresión de 26,6 a 24,7 cm (supramaleolar) y de 39,2 a 37,1 cm (infracondíleo; p < 0,001); puntuación media de la escala de dolor: inicial, 5,3 y final: 3,3 (p < 0, 001). b) Seguridad: 11 pacientes (9 por ciento) presentaron signos de sangrado, motivo de ingreso en uno. Seis (4,9 por ciento) presentaron algún tipo de reacción adversa, ninguna calificada como grave. En el 33,3 por ciento su relación con el fármaco se consideró como posible, y en el 66,7 por ciento, como muy probable. c) Aceptabilidad: el 82 por ciento de los pacientes se mostraron satisfechos con el tratamiento. Conclusiones. El estudio demuestra la eficacia y seguridad de la estrategia terapéutica que se llevó a cabo (AU)