Método y kit para la detección molecular de Tropheryma whipplei

La presente invención se refiere a un método para la detección de la bacteria Tropheryma whipplei en una muestra biológica aislada mediante la amplificación con pares de cebadores específicos de uno o más fragmentos de las secuencias nucleotídicas SEQ ID NO: 1, SEQ ID NO: 2 y SEQ ID NO: 3, donde dichas secuencias corresponden, respectivamente, a secuencias parciales de la secuencia nucleotídica del gen que codifica para la proteína HSP (Heat shock protein) de 65 kDa, de la secuencia nucleotídica de NCDRC (Non coding degenerated repeat cluster) y de la secuencia nucleotídica del gen que codifica para una proteína de la familia WISP (WNT1 inducible signaling pathway) de Tropheryma whipplei. Preferiblemente la amplificación se lleva a cabo por medio de PCR multiplex.; La detección de los productos de la amplificación se lleva a cabo por medio de sondas específicas complementarias a dichos productos. Asimismo, la presente invención se refiere a un kit que comprende los cebadores y sondas específicas para la detección de Tropheryma whipplei.REIVINDICACIONES: 1. Método para la detección de Tropher y ma whipplei que comprende: a. Obtener una muestra biológica aislada, b. poner la muestra del paso (a) , o parte de la misma, en contacto con una mezcla de reacción que contiene cebadores capaces de amplificar uno o más fragmentos de las secuencias nucleotídicas SEQ ID NO: 1, SEQ ID NO: 2 y SEQ ID NO: 3, c. amplificar los fragmentos de las secuencias nucleotídicas según el paso (b) mediante la reacción en cadena de la polimerasa, y d. detectar y/o cuantificar los productos de la amplificación del paso (c) . 2. Método según la reivindicación 1, donde los cebadores del paso (b) son: a. SEQ ID NO: 4 y SEQ ID NO: 5 para amplificar el fragmento SEQ ID NO: 1, b. SEQ ID NO: 6 y SEQ ID NO: 7 para amplificar el fragmento SEQ ID NO: 2, y c. SEQ ID NO: 8 y SEQ ID NO: 9 para amplificar el fragmento SEQ ID NO: 3. 3. Método según cualquiera de las reivindicaciones 1 ó 2, donde la amplificación se lleva a cabo por medio de PCR multiplex. 4. Método según cualquiera de las reivindicaciones 1 a 3, donde la detección y/o cuantificación se lleva a cabo por medio de una o más sondas moleculares capaces de hibridar con cualquiera de los fragmentos de las secuencias nucleotídicas. 5. Método según la reivindicación 4, donde las sondas moleculares para llevar a cabo la detección y/o cuantificación son: a. SEQ ID NO: 10 para detectar y/o cuantificar SEQ ID NO: 1, b. SEQ ID NO: 11 para detectar y/o cuantificar SEQ ID NO: 2, y c. SEQ ID NO: 12 para detectar y/o cuantificar SEQ ID NO: 3. 6. Método según cualquiera de las reivindicaciones 4 ó 5, donde además comprende sondas control capaces de hibridar con las secuencias modificadas de uno o más fragmentos de las secuencias nucleotídicas SEQ ID NO: 1, SEQ ID NO: 2 y SEQ ID NO: 3. 7. Método según la reivindicación 6, donde las sondas control, capaces de hibridar con las secuencias modificadas de dichos fragmentos, son SEQ ID NO: 13, SEQ ID NO: 14 y SEQ ID NO: 15. 8. Método según cualquiera de las reivindicaciones 4 a 7, donde la detección y/o cuantificación se lleva a cabo por medio de la hibridación de las sondas moleculares con las secuencias nucleotídicas amplificadas, en fase sólida. 9. Método según cualquiera de las reivindicaciones 1 a 8, donde la muestra biológica aislada es un fluido biológico o una biopsia de tejido biológico. 10. Método según cualquiera de las reivindicaciones 1 a 9, para la monitorización de la respuesta a un tratamiento de la enfermedad de Whipple, provocada por Tropher y ma whipplei. 11. Kit para la detección de Tropher y ma whipplei que comprende cebadores capaces de amplificar uno o más fragmentos de las secuencias nucleotídicas SEQ ID NO: 1, SEQ ID NO: 2 y SEQ ID NO: 3. 12. Kit según la reivindicación 11, donde los cebadores son: a. SEQ ID NO: 4 y SEQ ID NO: 5 para amplificar la secuencia nucleotídica SEQ ID NO: 1, b. SEQ ID NO: 6 y SEQ ID NO: 7 para amplificar la secuencia nucleotídica SEQ ID NO: 2, y c. SEQ ID NO: 8 y SEQ ID NO: 9 para amplificar la secuencia nucleotídica SEQ ID NO: 3. 13. Kit según cualquiera de las reivindicaciones 11 ó 12 que además comprende sondas moleculares capaces de hibridar con cualquiera de los fragmentos de las secuencias nucleotídicas. 14. Kit según la reivindicación 13, donde las sondas moleculares son SEQ ID NO: 10, SEQ ID NO: 11 y SEQ ID NO: 12. 15. Kit según cualquiera de las reivindicaciones 13 ó 14 que además comprende las sondas control SEQ ID NO: 13, SEQ ID NO: 14 y SEQ ID NO: 15, capaces de hibridar con secuencias modificadas de los fragmentos de dichas secuencias nucleotídicasCuando una patente se hace internacional, se puede encontrar en el idioma de cada país en que se ha solicitado. En Espacenet se tiene acceso a los documentos en cada idioma.Instituto de Salud Carlos IIISolicitud de patent

Bones pràctiques en atenció compartida: recomanacions per a una gestió òptima dels PIIC

Bones pràctiques; Presa decisions; Plans d’intervenció individualitzats i compartitsBuenas prácticas; Toma de decisiones; Planes de intervención individualizados i compartidosGood practices; Decision making; Individualized and shared intervention plansAquest document és un instrument per a professionals de l'atenció sanitària per posar al dia els criteris de bon ús dels PIIC respecte a la revisió que es va publicar l’abril de 2015

Molecular characterization of invasive serogroup B Neisseria meningitidis isolates from Spain during 2015-2018: Evolution of the vaccine antigen factor H binding protein (FHbp).

Studies of meningococcal genetic population structure, including the potential associations between surface proteins variants and clonal complexes, are important to understand how new protein MenB vaccines might impact in specific scenarios. With the aim to analyze the diversity of Spanish invasive MenB strains, and genetic variability of the fHbp vaccine antigen, all MenB isolates received at National Reference Laboratory (NRL) from 2015 to 2018 were molecularly characterized. 108, 103, 87 and 112 invasive MenB strains isolated during 2015-2018, respectively, were received at NRL. The strains were whole genome sequenced, and porA, fetA, MLST and fHbp variability was analyzed. Potential impact on MenB vaccines coverage was also assessed. A total of 42, 38 and 3 different FHbp subfamily A, B and A/B hybrid peptides, respectively, were found. FHbp subfamily A peptides were harboured by most of the strains (65.9%), being the most prevalent peptide 45 which was associated with genosubtype 22,14 and cc213. FHbp subfamily B peptides were harboured by 32.4% of the strains, and 6 strains harbouring subfamily A/B hybrid peptides were also found. The 64.15% of the strains showed FHbp variants "exact-match" or "cross-reactive" to the FHbp variants included in rLP2086 vaccine according to hSBA assays in the rLP2086 clinical development, and 15.85% showed FHbp peptides defined as predictors of FHbp-coverage for 4CMenB vaccine by gMATS. Due to invasive meningococcal strains temporal variability (eg prevalence of the cc213 increased from 3.6% in 2007 to 33% in 2018) affecting to the presence and distribution of the vaccine antigens, continuous detailed meningococcal surveillance and monitoring of the vaccine antigens is needed to determine the degree and durability of coverage provided by these protein vaccine.This study was partially supported by grants PI16CIII/00023 (MPY-1349/16) and PI19CIII/00030 (MPY-507/19) from Instituto de Salud Carlos III, and by Pfizer S.L.U. through an institutional agreement with the Instituto de Salud Carlos III (MVP-1273/16). This publication made use of the Neisseria Multi Locus Sequence Typing website (https://pubmlst.org/ neisseria/) sited at the University of Oxford (Jolley et al. Wellcome Open Res 2018, 3:124 [version 1; referees: 2 approved]). The development of this site has been funded by the Wellcome Trust and European Union. This study made use of Genomic and Bioinformatics ISCIII central Units for genomes sequencing and assembly. We thank all hospitals for sending the strains to the National Reference Laboratory for an IMD laboratory surveillance on a National level.S

Distribution of Bartonella henselae variants in patients, reservoir hosts and vectors in Spain

We have studied the diversity of B. henselae circulating in patients, reservoir hosts and vectors in Spain. In total, we have fully characterized 53 clinical samples from 46 patients, as well as 78 B. henselae isolates obtained from 35 cats from La Rioja and Catalonia (northeastern Spain), four positive cat blood samples from which no isolates were obtained, and three positive fleas by Multiple Locus Sequence Typing and Multiple Locus Variable Number Tandem Repeats Analysis. This study represents the largest series of human cases characterized with these methods, with 10 different sequence types and 41 MLVA profiles. Two of the sequence types and 35 of the profiles were not described previously. Most of the B. henselae variants belonged to ST5. Also, we have identified a common profile (72) which is well distributed in Spain and was found to persist over time. Indeed, this profile seems to be the origin from which most of the variants identified in this study have been generated. In addition, ST5, ST6 and ST9 were found associated with felines, whereas ST1, ST5 and ST8 were the most frequent sequence types found infecting humans. Interestingly, some of the feline associated variants never found on patients were located in a separate clade, which could represent a group of strains less pathogenic for humans.This work was supported by the Spanish Fondo de Investigación Sanitaria (PI10/00051 and PI10/00165). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.S

Rapid and Highly Sensitive DNA Flow Technology Platform to Detect Tick-Borne Bacterial Pathogens in Clinical Samples

Zoonotic diseases represent a significant public health concern worldwide due to the emergence/re-emergence of vector-borne diseases in the last decade. Ticks are the most important vectors in the northern hemisphere and can transmit diseases such as Lyme disease, human granulocytic anaplasmosis, and spotted fever rickettsioses, among others. Therefore, there is a growing need to develop better and faster diagnostic tools that can detect zoonotic human pathogens in clinical samples. In this study, we present the results for a new kit tick-borne bacteria flow chip (TBFC), which allows the simultaneous screening of seven different bacterial pathogens in human samples using a DNA flow technology platform (hybriSpot system). The analytical sensitivity and specificity of the TBFC were calculated spiking bacterial DNA in human DNA samples, and the results were compared with an in-house single PCR-reverse line blot (RLB) routinely used for diagnosis at the National Center for Microbiology in Spain. The analytical sensitivity and specificity of the TBFC were almost identical to the PCR-RLBs used in diagnosis. In addition, samples from patients (n = 212) with a wide range of clinical signs/symptoms consistent with multisystem disorders suggestive of a tick-borne infection were tested using the TBFC, and the results were compared with those obtained by PCR-RLB. The concordance of both methods using patient samples was 97.2%. The TBFC kit is a rapid new and cost-efficient diagnostic molecular tool capable of detecting tick-borne pathogens in clinical samples.S

Monkeypox virus genomic accordion strategies

The 2023 monkeypox (mpox) epidemic was caused by a subclade IIb descendant of a monkeypox virus (MPXV) lineage traced back to Nigeria in 1971. Person-to-person transmission appears higher than for clade I or subclade IIa MPXV, possibly caused by genomic changes in subclade IIb MPXV. Key genomic changes could occur in the genome's low-complexity regions (LCRs), which are challenging to sequence and are often dismissed as uninformative. Here, using a combination of highly sensitive techniques, we determine a high-quality MPXV genome sequence of a representative of the current epidemic with LCRs resolved at unprecedented accuracy. This reveals significant variation in short tandem repeats within LCRs. We demonstrate that LCR entropy in the MPXV genome is significantly higher than that of single-nucleotide polymorphisms (SNPs) and that LCRs are not randomly distributed. In silico analyses indicate that expression, translation, stability, or function of MPXV orthologous poxvirus genes (OPGs), including OPG153, OPG204, and OPG208, could be affected in a manner consistent with the established "genomic accordion" evolutionary strategies of orthopoxviruses. We posit that genomic studies focusing on phenotypic MPXV differences should consider LCR variability.We would like to thank the work of the Rapid Response Unit of the National Center for Microbiology, especially MªJosé Buitrago, and Cristobal Belda, ISCIII General Director. We also thank Anya Crane (Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health) for critically editing the manuscript and Jiro Wada (Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health) for helping with figure preparation. The work for this study performed at Instituto de Salud Carlos III was partially funded by Acción Estratégica “Impacto clínico y microbiológico del brote por el virus de la viruela del mono en pacientes en España (2022): proyecto multicéntrico MONKPOX-ESP22” (CIBERINFEC) (M.P.S.S.). The work for this study done at the Icahn School of Medicine at Mount Sinai Department of Microbiology as part of Global Health Emerging Pathogen Institute activities was funded by institutional funds (G.P.) from the Icahn School of Medicine at Mount Sinai Department of Microbiology in support of Global Health Emerging Pathogen Institute activities. This work was also supported in part through Laulima Govern ment Solutions, LLC, prime contract with the U.S. National Institute of Allergy and Infectious Diseases (NIAID) under Contract No. HHSN272201800013C. J.H.K. performed this work as an employee of Tunnell Government Services (TGS), a subcontractor of Laulima Government Solutions, LLC, under Contract No. HHSN272201800013C. Opinions, interpretations, conclusions, and recommendations are those of the authors and are not necessarily endorsed by the U.S. Army. The views and conclusions contained in this document are those of the authors and should not be interpreted as necessarily representing the official policies, either expressed or implied, of the U.S. Department of Health and Human Services or of the institutions and companies affiliated with the authors, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government.S

Association of irisin with fat mass, resting energy expenditure, and daily activity in conditions of extreme body mass index

FNDC5/irisin has been recently postulated as beneficial in the treatment of obesity and diabetes because it is induced in muscle by exercise, increasing energy expenditure. However, recent reports have shown that WAT also secretes irisin and that circulating irisin is elevated in obese subjects. The aim of this study was to evaluate irisin levels in conditions of extreme BMI and its correlation with basal metabolism and daily activity. The study involved 145 female patients, including 96 with extreme BMIs (30 anorexic (AN) and 66 obese (OB)) and 49 healthy normal weight (NW). The plasma irisin levels were significantly elevated in the OB patients compared with the AN and NW patients. Irisin also correlated positively with body weight, BMI, and fat mass. The OB patients exhibited the highest REE and higher daily physical activity compared with the AN patients but lower activity compared with the NW patients. The irisin levels were inversely correlated with daily physical activity and directly correlated with REE. Fat mass contributed to most of the variability of the irisin plasma levels independently of the other studied parameters. Conclusion. Irisin levels are influenced by energy expenditure independently of daily physical activity but fat mass is the main contributing factor