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Investigation

Mechanisms of Antifungal Resistance in Aspergillus

Research Lines

Content with Investigacion Mecanismos de resistencia a antifúngicos en Aspergillus .

Resistance mechanisms of Aspergillus fumigatus to antifungals

Aspergillus fumigatus is a universally distributed opportunistic fungal pathogen with a significant global incidence and extremely high mortality. The widespread and extensive use of azole antifungals has led to the emergence of A. fumigatus azole resistant, resulting in serious consequences for patients infected with these isolates, who are left with limited therapeutic options.       
Initially, the emergence of resistant strains was very sporadic and showed point mutations in key areas of the Cyp51A enzyme (G54, G138, F219, M220, G448S) in strains isolated from patients undergoing long-term treatment with azoles. This clinical pathway is due to the selective pressure that azoles exert on A. fumigatus within the patient. However, since 2014, resistance has grown significantly, and almost all azole-resistant A. fumigatus strains have a combined mechanism of modifications in the promoter and the coding portion of cyp51A (TR34/L98H or TR46/Y121F/T289A). Both resistance mechanisms are frequently detected in strains from patients who have never been exposed to antifungal therapy. In these cases, the involvement of an environmental route is raised, in which the unintentional exposure of A. fumigatus to DMIs (imidazole and triazole) in the field would be favoring the resistance emergence.

Origin and Evolution of A. fumigatus Resistance to Antifungals

Nowadays, the isolation of A. fumigatus strains resistant to antifungals is an increasing global emergence. The continuous exposure of A. fumigatus to environmental fungicides, used for crop protection against other fungal species that cause agricultural damage, is believed to be selecting multi drug resistant strains. The main azole resistance mechanisms in A. fumigatus are strains with modifications of the azole target (cyp51A gene), mainly the TR34/L98H, followed by TR46/Y121F/A289T. Both types of mechanisms are responsible for panazole resistance and cross resistance to DMIs used for crop protection (imidazoles and triazoles). More recently, resistance to several fungicide classes such as, Bencimidazoles (MBC), Estrobilurinas (QoIs), sucinato deshidrogenase inhibitors (SDHIs) and  Dicarboximides, has also been acknowledged.

Genomic characterization (NGS) of strains from both clinical and environmental sources allows linking genomic differences with the acquisition of resistance to different fungicides. Adding data on susceptibility to non-azole antifungals provides a more precise picture of the phylogenetic relationships among strains, as distinct subclades are formed in which strains multi-resistant to non-azole antifungals grouped with azole-resistant strains with TRs resistance mechanisms. This formation of specific clades with strains that differ in geographic origin and year of isolation suggests the existence of a common link, an evolutionary origin according to which the strains have developed under similar circumstances that converge in a series of multi-resistance mechanisms to fungicides from different families. The resistance of A. fumigatus to non-azole fungicides, that are exclusively used in the environment, confirms that the strains with TRs resistance mechanisms are selected and developed in the environment where they are exposed to the selective pressure of multiple fungicides.

Tolerance and Persistence to Azole Antifungals in Aspergillus fumigatus

Tolerance and persistence are two phenomena by which pathogenic organisms can survive the microbicidal action of antimicrobials that should kill them over an extended period. In our laboratory, we investigate the ability of certain A. fumigatus isolates to exhibit tolerance and persistence to azoles, which are the first-line antifungal treatment for aspergillosis infections.

We are developing methodologies to detect and study tolerance and persistence, both in the laboratory and in clinical diagnosis. Using these methods, we are exploring the underlying molecular and genomic mechanisms that enable these phenomena. In addition, we are investigating the potential relevance of tolerance and persistence in the efficacy of antifungal treatment.

Differential Modulation of Persulfidation in the Fungus and Host as a Novel Antifungal Strategy

Persulfidation is a post-translational modification in which an activated sulfur group (S₂-), through the action of an enzyme, performs a specific nucleophilic attack on thiol (-SH) groups of cysteine residues in target proteins, forming a persulfide group (-SSH). This modification has been shown to modulate the intrinsic activity of proteins, playing a crucial role in various cellular mechanisms and physiological functions.

In our previous research, we demonstrated that correct levels of persulfidation are important both for A. fumigatus virulence and for orchestrating an adequate immune response in the host. Based on this, our research explores the hypothesis that differential modulation of persulfidation could constitute a novel antifungal treatment strategy.

We are investigating the ability of compounds to inhibit fungal enzymes responsible for persulfidation, aiming to reduce persulfidation levels and thereby decrease A. fumigatus virulence. Additionally, we are studying the use of sulfur donors as a potential means to enhance persulfidation in pulmonary host cells, with the goal of strengthening the immune response.

Evolution of Cross-Resistance to the New Antifungals Olorofim and Manogepix

Azole resistance is already present worldwide. Studies have shown that the most common resistance mechanisms—tandem repeats in the promoter of the gene encoding the azole target—have developed in agricultural settings due to the indiscriminate use of pesticides from the same family as clinical azoles.

Currently, two new clinical antifungals with novel molecular mechanisms of action have been introduced: olorofim and manogepix. However, analogous compounds with the same mechanism of action, ipflufenoquin and aminopyrifen, have also been developed for use as pesticides. This situation puts us at risk of repeating the same mistake made with azoles.

In this international collaborative project, we study the evolution of resistance and cross-resistance to these clinical and environmental antifungals. Our goal is to design strategies to minimize the emergence of resistance in the environment and develop early detection methods for antifungal resistance.

Research projects

Content with Investigacion Mecanismos de resistencia a antifúngicos en Aspergillus .

PROJECT TITLE: Consorcio Centro de Investigacion Biomedica en Red (CIBER).  Infectious Diseases Area. 
Funding Agency: CIF: G85296226.  Reference: CB21/13/00105
Dates: 2022-2026            Funding: 85.000 € (first year)
Principal Investigator: Emilia Mellado Terrado 


 

PROJECT TITLE: Modulación diferencial de la persulfidación en el hongo y el hospedador como nueva estrategia antifúngica. 
Funding Agency: Agencia Estatal de Investigación (Convocatoria Proyectos de Generación de Conocimiento"
Reference: Project PID2022-136343OA-I00 funded by MICIU/AEI /10.13039/501100011033 and by FEDER, UE
Principal Investigator: Jorge Amich. 
Dates: 2024-2026. 
Funding: 118.750 €


 

PROJECT TITLE: : Bridging the gap between environment and patient JPIAMR (AC23CIII_2/00002 (JPIAMR2023-DISTOMOS-103). 
DATES: 2024-2026            Funding: 178.000 €
Principal Investigator: Jorge Amich. 

PROJECT TITLE: : Buscando los rasgos geneticos de la resistencia de Aspergillus fumigatus a los azoles para preservar la eficacia de los azoles:un enfoque de salud global.
FUNDING AGENCY: Fondo de Investigación Sanitaria. PI21CIII/00028_ MPY443/2021
DATES: 2022-2025            Funding: 47.000 €
Principal Investigator: Emilia Mellado Terrado 

PROJECT TITLE: : Persistencia a antifúngicos azólicos en Aspergillus fumigatus: mecanismos, relevancia y diagnóstico. 
FUNDING AGENCY: AESI 2022 (PI22CIII/00053). 
DATES: 2023-2025            Funding: 55.000 €
Principal Investigator: Jorge Amich. 

PROJECT TITLE: : La medicina de precisión contra la resistencia a antimicrobianos:
CONSORCIO CENTRO DE INVESTIGACION BIOMEDICA EN RED (CIBER) CENTRO NACIONAL DE MICROBIOLOGIA
G85296226 PMP22/00092. Project MePRAM 28.107.46QF.749   Funding: 4.339.500,00€
Principal Investigator: Jesus Oteo 

Publications

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Multidisciplinary Group of viral coinfection HIV/Hepatitis (COVIHEP). HCV-coinfection is related to an increased HIV-1 reservoir size in cART-treated HIV patients: a cross-sectional study.

López-Huertas MR, Palladino C, Garrido-Arquero M, Esteban-Cartelle B, Sánchez-Carrillo M, Martínez-Román P, Martín-Carbonero L, Ryan P, Domínguez-Domínguez L, Santos ID, Moral SDLF, Benito JM, Rallón N, Alcamí J, Resino S, Fernández-Rodríguez A, Coiras M, Briz V*, on behalf of the Multidisciplinary Group of viral coinfection HIV/Hepatitis (COVIHEP). HCV-coinfection is related to an increased HIV-1 reservoir size in cART-treated HIV patients: a cross-sectional study. Sci Rep 2019: 9 (1); 5606.

PUBMED DOI

Persistent Low-Level Viremia in treatment HIV infection: An unresolved status.

3. Crespo-Bermejo C, Ramírez de Arellano E, Lara-Aguilar V, Martínez-Román P, Arca Lafuente S, Gómez-Lus ML, Martín-Carbonero L, Briz V. Persistent Low-Level Viremia in treatment HIV infection: An unresolved status. Virulencia 2021, 12: 2919-2931.

PUBMED DOI

Full coding hepatitis E virus genotype 3 genome amplification method.

Muñoz-Chimeno M, Forero JE, Echevarría JM, Muñoz-Bellido JL, Vázquez-López L, Morago L, García-Galera MC, Avellón A. Full coding hepatitis E virus genotype 3 genome amplification method. J Virol Methods. 2016 Apr;230:18-23.

PUBMED DOI

Comparative sensitivity of commercial tests for hepatitis E genotype 3 virus antibody detection.

• Avellón A, Morago L, García-Galera del Carmen M, Muñoz M, Jose-Manuel Echevarría JM. Comparative sensitivity of commercial tests for hepatitis E genotype 3 virus antibody detection. J Med Virol. 2015 Nov;87(11):1934-9.

PUBMED DOI

Epidemiology and predictive factors for early and late mortality in Candida bloodstream infections: a population-based surveillance in Spain

11: Puig-Asensio M, Padilla B, Garnacho-Montero J, Zaragoza O, Aguado JM, Zaragoza R, Montejo M, Muñoz P, Ruiz-Camps I, Cuenca-Estrella M, Almirante B; CANDIPOP Project; GEIH-GEMICOMED (SEIMC); REIPI. Epidemiology and predictive factors for early and late mortality in Candida bloodstream infections: a population-based surveillance in Spain. Clin Microbiol Infect. 2014 Apr;20(4):O245-54.

PUBMED DOI

B Pérez de Val, B Romero, MT Tórtola, L Herrera-León, P Pozo, I Mercader, JL Sáez, M Domingo, E Vidal. Poly-resistant Mycobacterium bovis infection in a human and sympatric sheep, Spain, 2017-2018

B Pérez de Val, B Romero, MT Tórtola, L Herrera-León, P Pozo, I Mercader, JL Sáez, M Domingo, E Vidal. Poly-resistant Mycobacterium bovis infection in a human and sympatric sheep, Spain, 2017-2018. Emerg Infect Dis. 2021 Apr;27(4):1241-1243. doi: 10.3201/eid2704.204467. PMID: 33755008.

DOI

L Bernal-Martínez; L Herrera-Leon; C Valero; P de la Cruz; L Ghimpu; AC Mesa-Arango; G Santoni; L Goterris; R Millán; MJ Buitrago. Differential Diagnosis of Fungal Pneumonias vs.Tuberculosis in AIDS Patients by Using Two New Molecular Methods.

L Bernal-Martínez; L Herrera-Leon; C Valero; P de la Cruz; L Ghimpu; AC Mesa-Arango; G Santoni; L Goterris; R Millán; MJ Buitrago. Differential Diagnosis of Fungal Pneumonias vs.Tuberculosis in AIDS Patients by Using Two New Molecular Methods. J. Fungi 2021, 7, 336. doi.org: 10.3390/jof7050336. PMID: 33925404.

DOI

E Tagliani, RAnthony, TA Kohl, A de Neeling, V Nikolayevskyy, C Ködmön, FP Maurer, S Niemann, D van Soolingen, MJ van der Werf, D Cirillo, ECDC molecular surveillance project participants. Use of a whole genome sequencing-based approach for Mycobacterium tuberculosis surveillance in Europe in 2017-2019: an ECDC pilot study

E Tagliani, RAnthony, TA Kohl, A de Neeling, V Nikolayevskyy, C Ködmön, FP Maurer, S Niemann, D van Soolingen, MJ van der Werf, D Cirillo, ECDC molecular surveillance project participants. Use of a whole genome sequencing-based approach for Mycobacterium tuberculosis surveillance in Europe in 2017-2019: an ECDC pilot study. Eur Respir J. 2021 Jan 5;57(1):2002272. doi: 10.1183/13993003.02272-2020. Print 2021 Jan. PMID: 32732329.

DOI

MJ Iglesias, D Ibarz, A Cebollada, J Comín, MS Jiménez, MC Vázquez, S Samper, Spanish Working Group on MDRTB. The value of the continuous genotyping of multidrug resistant tuberculosis over 20 years in Spain.

MJ Iglesias, D Ibarz, A Cebollada, J Comín, MS Jiménez, MC Vázquez, S Samper, Spanish Working Group on MDRTB. The value of the continuous genotyping of multidrug resistant tuberculosis over 20 years in Spain. Sci Rep. 2020 Nov 24;10(1):20433. doi: 10.1038/s41598-020-77249-x. PMID: 33235225.

DOI

S Campos-Gutierrez, MJ Ramos-Real, R Abreu, MS Jimenez, M Lecuona. Pseudo-ourbreak of Mycobacterium fortuitum, in a hospital bronchoscopy unit.

S Campos-Gutierrez, MJ Ramos-Real, R Abreu, MS Jimenez, M Lecuona. Pseudo-ourbreak of Mycobacterium fortuitum, in a hospital bronchoscopy unit. Am J Infect Control. 2020 Jul;48(7):765-769. doi: 10.1016/j.ajic.2019.11.019. Epub 2019 Dec 24. PMID: 31882175.

DOI

Gascha , Y Meijeb, M Espasac, B Fonta, MS Jiménez, N Fernández-Hidalgo. Disseminated Infection Due to Mycobacterium chimaera After Aortic Valve Replacement.

Gascha , Y Meijeb, M Espasac, B Fonta, MS Jiménez, N Fernández-Hidalgo. Disseminated Infection Due to Mycobacterium chimaera After Aortic Valve Replacement. Revista Española de cardiología. 2019. Vol 72 (6):502-503. DOI: 10.1016/j.rec.2018.06.026. PMID: 30029979

DOI

PBMCs gene expression signature of advanced cirrhosis with high risk for clinically significant portal hypertension in HIV/HCV coinfected patients: A cross-control study

2. Salgüero S, Brochado-Kith O, Virseda Verdices A, Berenguer J, González-García J, Martínez I, Díez C, Hontañón V, Pérez-Latorre L, Fernández-Rodríguez A (‡), Jiménez-Sousa MA (‡,*), and Resino S (‡, *). PBMCs gene expression signature of advanced cirrhosis with high risk for clinically significant portal hypertension in HIV/HCV coinfected patients: A cross-control study. Biomed Pharmacother 2023, 159:114220. (A; FI= 7.42; D1, Pharmacology & Pharmacy; JCR 2021). PMID: 36628818. DOI: 10.1016/j.biopha.2023.114220.

PUBMED

Low anti-SARS-CoV-2 S antibody levels predict increased mortality and dissemination of viral components in the blood of critical COVID-19 patients

4. Martin-Vicente M, Almansa R, Martínez I, Tedim AP, Bustamante E, Tamayo L, Aldecoa C, Gómez JM, Renedo G, Berezo JA, Cedeño JA, Mamolar N, García Olivares P, Herrán-Monge R, Cicuendez R, Enríquez P, Ortega A, Jorge N, Doncel C, Fuente A, Bustamante-Munguira J, Muñoz-Gómez MJ, González-Rivera M, Puertas C, Más V, Vázquez M, Pérez-García F, Rico-Feijoo J, Martín S, Motos A, Fernandez-Barat L, Eiros JM, Domínguez-Gil M, Ferrer R, Barbé F, Trapiello W, Kelvin DJ (¥), Bermejo-Martin JF (* ¥), Resino S (¥), Torres A (* ¥). Low anti-SARS-CoV-2 S antibody levels predict increased mortality and dissemination of viral components in the blood of critical COVID-19 patients. J Intern Med. 2022, 291(2):232–240. (A; FI= 13.07; D1, Medicine, General & Internal; JCR 2021). PMID: 34611927. DOI: 10.1111/joim.13386.

PUBMED

Prevalence and factors associated with SARS-CoV-2 seropositivity in the Spanish HIV Research Network Cohort

5. Berenguer J (*), Díez C, Martín-Vicente M, Mican R, Pérez-Elías MJ, García-Fraile LJ, Vidal F, Suárez-García I, Podzamcer D, Del Romero J, Pulido F, Iribarren JA, Gutiérrez F, Poveda E, Galera C, Izquierdo R, Asensi V, Portilla J, López JC, Arribas JR, Moreno S, González-García J, Resino S (‡), Jarrín I (‡). Prevalence and factors associated with SARS-CoV-2 seropositivity in the Spanish HIV Research Network Cohort. Clin Microbiol Infect. 2021, 27(11):1678-1684. (A; FI= 13.31; D1, Microbiology; JCR 2021). PMID: 34186209 DOI: 10.1016/j.cmi.2021.06.023.

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Environmental factors linked to hospital admissions in young children due to acute viral lower respiratory infections: A bidirectional case-crossover study

6. Álvaro-Meca A, Goez MC, Resino R, Matías V, Sepúlveda-Crespo D; Martínez I, Resino S (*). Environmental factors linked to hospital admissions in young children due to acute viral lower respiratory infections: a bidirectional case-crossover study. Environ Res. 2022. 212(Pt B), 113319. (A; FI= 8.43; D1, Public, Environmental & Occupational Health; JCR 2021). PMID: 35447151. DOI: 10.1016/j.envres.2022.113319.

PUBMED

Detection of active hepatitis C in a single visit and linkage to care among marginalized people using a mobile unit in Madrid, Spain

7. Ryan P, Valencia J, Cuevas G, Torres-Macho J, Troya J, Pueyo A, Muñoz-Gómez MJ, Muñoz-Rivas N, Vázquez-Morón S, Martínez, I, Lazarus JV, Resino S (*). Detection of active hepatitis C in a single visit and link-to-care among excluded people using a mobile unit in Madrid, Spain. Int J Drug Policy. 2021, 96:103424. (A; FI= 5.93; Q1, Substance Abuse; JCR 2021). PMID: 34429222 DOI: 10.1016/j.drugpo.2021.103424.

PUBMED

María José Ruiz-Lopéz, Milagros Muñoz-Chimeno, Jordi Figuerola, Ana M. Gavilán, Sarai Varona, Isabel Cuesta, Josué Martínez-de la Puente, Angel Zaballos, Francisca Molero, Ramón C. Soriguer, Mª Paz Sánchez-Seco, Santiago Ruiz, Ana Vázquez. Genomic analysis of West Nile virus lineage 1 isolated from mosquitoes

María José Ruiz-Lopéz, Milagros Muñoz-Chimeno, Jordi Figuerola, Ana M. Gavilán, Sarai Varona, Isabel Cuesta, Josué Martínez-de la Puente, Angel Zaballos, Francisca Molero, Ramón C. Soriguer, Mª Paz Sánchez-Seco, Santiago Ruiz, Ana Vázquez. Genomic analysis of West Nile virus lineage 1 isolated from mosquitoes from 2020-2021 outbreak occurred in Andalusia, Spain. Viruses 2023, 15, 266. https://doi.org/10.3390/v15020266

DOI

Jordi Figuerola, Miguel Ángel Jiménez-Clavero, María José Ruíz-López, Francisco Llorente, Santiago Ruiz, Andreas Hoefer, Pilar Aguilera-Sepúlveda, Jéssica Jiménez Peñuela1, Olaya García-Ruiz, Laura Herrero, Ramón C. Soriguer, Raúl Fernández Delgado, Mari Paz Sánchez-Seco, Josué Martínez-de la Puente, Ana Vázquez. A One Health view of the West Nile virus outbreak in Andalusía (Spain) in 2020

Jordi Figuerola, Miguel Ángel Jiménez-Clavero, María José Ruíz-López, Francisco Llorente, Santiago Ruiz, Andreas Hoefer, Pilar Aguilera-Sepúlveda, Jéssica Jiménez Peñuela1, Olaya García-Ruiz, Laura Herrero, Ramón C. Soriguer, Raúl Fernández Delgado, Mari Paz Sánchez-Seco, Josué Martínez-de la Puente, Ana Vázquez. A One Health view of the West Nile virus outbreak in Andalusía (Spain) in 2020. Emerg Microbes Infect. 2022 Dec;11(1):2570-2578. doi: 10.1080/22221751.2022.2134055.

DOI

HCV eradication with IFN-based therapy does not completely restore gene expression in PBMCs from HIV/HCV-coinfected patients.

10.1186/s40249-021-00894-5. 9. Brochado O, Martínez I (*), Berenguer J, Medrano L, González-García J, Jiménez-Sousa MA, Carrero A, Hontañón V, Navarro J, Guardiola JM, Pérez-Latorre L, Micán R, Fernández-Rodríguez A (‡), Resino S (* ‡). HCV eradication with IFN-based therapy does not completely restore gene expression in PBMCs from HIV/HCV-coinfected patients. J Biomed Sci 2021; 28:23 (A; FI= 12.77; D1, Medicine, Research & Experimental; JCR 2021). PMID: 33785040. DOI: 10.1186/s12929-021-00718-6.

PUBMED

Relationship of vitamin D status with advanced liver fibrosis and response to hepatitis C virus therapy: A meta-analysis.

12. García-Álvarez M, Pineda-Tenor D, Jiménez-Sousa Ma, Fernández-Rodríguez A, Guzmán-Fulgencio M; Resino S (*). Relationship of vitamin D status with advanced liver fibrosis and response to hepatitis C virus therapy: A meta-analysis. Hepatology. 2014, 6(5):1541-1550. (A; FI= 11.05; D1, Gastroenterology & Hepatology; JCR 2014). PMID: 24975775. DOI: 10.1002/hep.27281.

PUBMED

List of staff

Additional Information

The current director of CNM is Dr. José Miguel Rubio Muñoz.

Dr. José Miguel Rubio has a degree in Biological Sciences from the Universidad Autónoma de Madrid (1986) and a PhD in Biological Sciences from the same university (1992). He carried out his doctoral thesis at the Department of Genetics of the Universidad Autónoma de Madrid, as Associate Professor (1988-1989), and at the School of Biology of the University of East Anglia in Norwich, UK, as Senior Research Assistant (1989-1992).

During his postdoctoral period he obtained a grant from the European Commission within the Human Capital and Mobility Program to be carried out at the University of “La Sapienza” in Rome, Italy and the Institute of Molecular Biology and Biotechnology in Crete, Greece (1993-1994). Subsequently, he made a further stay funded by the WHO and the university itself at the Department of Entomology, Wageningen University, The Netherlands (1994-1996).

Since 1997 he has been a member of the Instituto de Salud Carlos III (ISCIII), where he joined the Department of Parasitology of the National Center of Microbiology, as an EU-INCO postdoctoral fellow and later with a grant from the Autonomous Community of Madrid (CAM). She was part of the founding group of the National Center for Tropical Medicine (2003-2006) and of the 24/7 Alerts and Emergencies Unit (2006-2018) and is currently Head of the Malaria and Emerging Parasitosis Unit of the National Microbiology Center and is part, as research staff, of the Center for Biomedical Research Network on Infectious Diseases (CIBERINFEC/ISCIII).

During his scientific career he has been Visiting Scientist at the Leonidas e Marie Dean Center (FIOCRUZ-AMAZONAS, Manaus, Brazil) and is an External Consultant of the Parasitology Departments of Cairo University (Egypt) and the Medical Research Center (MRC) of Kuala Lumpur (Malaysia).  He also belongs or has belonged to different national and international committees:  Member of the expert group for malaria control of the European Centre for Disease Control (ECDC) since 2011; Expert-Evaluator for health programs of the European Commission since 2004; Spanish Representative (commissioned by ISCIII and MSC) in the Technical Scientific Committee of the TDR (WHO) 2007-2008; Spanish Deputy Focal Point for microbiology at the European Centre for Disease Control (ECDC) from 2012 to 2020; and, member of the Research Ethics Committee of ISCIII until 2019.

In this period he has published more than 100 articles in international indexed journals, 10 book chapters and has been co-editor of two books in the area of malaria, tropical medicine and neglected diseases. He has participated in 58 competitively funded research projects, 20 of them international, having been the principal investigator in 8 national and 11 international projects as PI of the project or WP leader. In addition, he has led five agreements with companies. Currently he has been awarded four sexenios of research, being presented this year 2025 to the fifth. In the teaching field, he participates in different postgraduate programs in the areas of microbiology and parasitology, having directed seven doctoral theses and more than 20 Master's or Degree final projects, both nationally and internationally. ​​​​​

El laboratorio de Referencia e Investigación en Resistencia a Antibióticos ofrece una amplia cartera de servicios al Sistema Nacional de Salud, las cuales pueden solicitarse en cnm-laboratorios.isciii.es. Jefe del Laboratorio: Jesús Oteo Iglesias (Punto focal Nacional de Resistencia antibiótica).

Dispone de dos programas de Vigilancia oficiales y gratuitos que engloban los ensayos ofertados ya sea como aislamientos individuales o mediante estudio de brotes. El Laboratorio utiliza asimismo técnicas de PCR en tiempo real para la detección de genes de resistencia, estas técnicas se han adaptado a un formato multiplex que permite detectar varios genes en la misma reacción. En los últimos años se han incluido metodologías basadas en la secuenciación de genomas completos para el análisis de bacterias multiresistentes (WGS).

Programa de vigilancia de Haemophilus influenzae. Responsables: María Pérez Vázquez (Punto focal Nacional de Haemophilus influenzae) y Belén Aracil. Laboratorio encargado de la identificación, estudio de sensibilidad y análisis genotípico de aislados de Haemophilus influenzae, centrándose esencialmente en la patología invasiva debida este patógeno. 

Programa de vigilancia de Resistencia a Antibióticos. Responsables: María Pérez Vázquez  y Belén Aracil (Punto focal Nacional de Resistencia antibiótica). Laboratorio encargado de la identificación, el estudio de sensibilidad antibiótica, y el diagnóstico fenotípico y genotípico de los diferentes mecanismos de resistencia a antibióticos fundamentalmente en enterobacterias y gram-negativos no fermentadores y Enterococcus spp.

Estudio de brotes. Responsables: Belén Aracil y María Pérez Vázquez. El programa incluye la caracterización de brotes nosocomiales y clones emergentes de alto riesgo mediante diferentes técnicas moleculares (tabla resumen). Éstas, nos permiten realizar estudios filogenéticos con el fin de obtener una información detallada acerca la relación entre los diferentes aislados y su trazabilidad. El objetivo final es generar datos que se transfieren a los hospitales como ayuda para la prevención o control de la propagación del brote.

Acreditación y Calidad. Responsable: Belén Aracil. El laboratorio Referencia e Investigación en Resistencia a Antibióticos ha sido de los primeros en el ISCIII en la utilización de técnicas acreditadas por la Entidad Nacional de Acreditaciones (ENAC). Este laboratorio consiguió la primera acreditación homologada de técnicas diagnósticas en 2012, programa que ha sido ampliado, de manera que en la actualidad más de la mitad de las técnicas ofrecidas al Sistema Nacional de Salud están debidamente acreditadas por ENAC.

Técnicos responsables de las técnicas realizadas en el Laboratorio: Noelia Lara Fuella y Verónica Bautista Sánchez.

En la siguiente imagen se resumen las técnicas ofrecidas al Sistema Nacional de Salud.

PROGRAMAS NOMBRE CARTERA SERVICIO PATÓGENO DETERMINACIÓN, DETECCIÓN, ANÁLISIS MÉTODOS

Programa de vigilancia de Haemophilus

Programa de vigilancia de resistencia a antibióticos.

Identificación bacteriana

Haemophilus sp.

Enterobacterias, gram-negativos no fermentadores, Enterococcus spp

Identificación bacteriana

Bioquímicos

MALDI TOF

Secuenciación de RNAr

Identificación capsular

Haemophilus influenzae

 

Identificación capsular fenotípica y genotípica

Aglutinación serológica en latex

PCR ind/multiplex

Determinación de Sensibilidad

Haemophilus sp.

Enterobacterias, gram-negativos no fermentadores, Enterococcus

 

Determinación de Sensibilidad

Microdilución                

Tiras epsilon               

Kirby Bauer

Métodos fenotípicos de detección de mecanismos de resistencia

Enterobacterias, gram-negativos no fermentadores,

 

Métodos fenotípicos de detección de mecanismos de resistencia

Discos y tabletas combinados con inhibidores                

Tiras combinadas     

Test de Hodge modificado

CabaNP                               

Inmunocromatografía CBP

Métodos genotípicos de detección de mecanismos de resistencia

Haemophilus sp.

Enterobacterias, gram-negativos no fermentadores, Enterococcus

 

ADN, PCR y secuenciación

PCR ind/multiplex

Análisis comparativo de las secuencias

Tipificación molecular/análisis filogenéticos

Haemophilus sp.

Enterobacterias, gram-negativos no fermentadores, Enterococcus

 

Corte enzimas de restricción, electroforesis

ADN, PCR y secuenciación

Preparación de librerías y secuenciación y análisis de genomas completos

 

PFGE

 

MLST

 

WGS