Centro Nacional de Microbiología (CNM)

Permanent URI for this collectionhttps://hdl.handle.net/20.500.12105/19609

El Centro Nacional de Microbiología (CNM) es uno de los centros pertenecientes al Instituto de Salud Carlos III que proporciona apoyo científico-técnico a la Administración General del Estado, a las Comunidades Autónomas y al Sistema Nacional de Salud (SNS), tal y como se recoge en La Ley General de Sanidad (Ley 14/1986, de 25 de abril) y el Estatuto del Instituto de Salud Carlos III (RD 375/2001, de 6 abril y su posterior reforma, RD 1672/2009, de 6 de noviembre). La función específica del CNM es el control de las enfermedades infecciosas para lo que ofrece servicios de diagnóstico y referencia, manteniendo además programas de investigación, tanto básica como orientada, relacionados con la prevención, el diagnóstico y el tratamiento de estas enfermedades. Dispone de un sistema de gestión de calidad conforme a la norma UNE-EN ISO 9001, certificado por AENOR, para la recepción de muestras biológicas, así como varias técnicas y servicios acreditados por ENAC según la norma UNE-EN ISO 15189.

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Heat-inactivated Mycobacterium bovis and P22PI protein immunocomplex: Two candidates for use as immunostimulants of innate immune response.
(Elsevier, 2025-06) Agulló-Ros, Irene; Burucúa, Mercedes M; Cheuquepán, Felipe A; Dominguez-Rodriguez, Mercedes; Sevilla, Iker A; Martínez, Remigio; Plá, Natalia; Risalde, María A; Marin, Maia S; University of Córdoba (España); Ministerio de Ciencia, Innovación y Universidades (España); Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF); Agencia Nacional de Promoción de la Investigación, el Desarrollo Tecnológico y la Innovación
Tuberculosis (TB), caused by members of the Mycobacterium tuberculosis complex, remains a critical global health challenge, affecting humans and a wide range of domestic and wild animals. Despite the availability of anti-TB drugs, cure rates remain suboptimal, exacerbated by the rise of multidrug-resistant TB strains. The Bacille Calmette-Guérin (BCG) vaccine, the only licensed vaccine against TB, has demonstrated efficacy in reducing lesion severity and bacterial burden in animals, as well as lowering TB-related and all-cause mortality in infants. However, BCG presents several safety concerns inherent to live vaccines. To overcome these limitations, exploring alternative vaccine candidates that do not incorporate live mycobacteria is crucial. This study aimed to evaluate and compare the immunostimulatory potential of two candidates based in mycobacteria inactivated or their derivatives, heat-inactivated Mycobacterium bovis (HIMB) and P22PI protein immunocomplex (P22PI), in bovine foetal lung cells. To assess the expression of innate immune components, including Toll-like receptors (TLRs), cathelicidins, and cytokines, bovine foetal lung were exposed to different concentrations of HIMB and P22PI immunostimulants, starting at 7.8 × 10⁶ CFU/ml and 10 µg/ml, respectively. These initial concentrations were subsequently diluted to 1/2 and 1/10 to evaluate dose-dependent effects. Our findings reveal that both HIMB and P22PI significantly stimulate innate immune mechanisms, as evidenced by the upregulation of TLR2 and TLR4, alongside the induction of BMAP28 cathelicidin, tumour necrosis factor alpha (TNFA) and interferons (IFNs). These results suggest their potential to orchestrate a robust innate immune response providing valuable insights into the immunological mechanisms underlying the protective effects of these immunostimulants. This underscores their potential role in in vivo studies as vaccine candidates. Furthermore, their ability to enhance antigen recognition via TLR and induce pro-inflammatory cytokines also indicates broader applications in immune modulation, potentially extending protection against heterologous pathogens through trained immunity.
Trends in Paediatric Viral Meningitis and Encephalitis With Unconfirmed Aetiology: A Spanish Population-Based Study, 2016-2020.
(Wiley, 2025-09) Pons-Espinal, Marina; Lopez-Perea, Noemi; Masa-Calles, Josefa; Muñoz-Almagro, Carmen; Tarrago Asensio, David; Launes, Cristian; Centro de Investigación Biomédica en Red - CIBERESP (Epidemiología y Salud Pública)
Aim: The prevalence of meningitis and encephalitis of unknown aetiology in Spanish children has not been specifically documented before. The aim of this study is to describe the epidemiology and trends of these clinical conditions between 2016 and 2020. Methods: Retrospective study analysing hospitalised children < 15 years with meningitis and encephalitis/encephalomyelitis of unknown origin (ICD-10 codes) in Spain (2016-2020). Data from National Registry of Hospitalizations and National Institute for Statistics were used to calculate age-stratified hospitalisation rates (HR). Statistical analysis included Poisson regression to calculate hospitalisation rate ratios (HRR) by age groups and years and forecasting methods to predict 2020 HRs. Results: Four thousand six hundred childrens were hospitalised with viral meningitis and encephalitis-encephalomyelitis of unknown origin, resulting in a HR of 7.8/105 inhabitants. The highest HR was observed in children under 1 year (49.1/105) and those aged 5-9 (10.4/105). The global HR for viral meningitis (3.64/105) was lower than for encephalitis-encephalomyelitis (4.2/105). Hospitalisations decreased from 1475 (2016) to 452 (2020), attributed to enhanced pathogen detection methods and COVID-19 preventive measures. Conclusion: Undiagnosed central nervous system entities remain a significant cause of paediatric hospitalisations in Spain, despite a declining incidence. Enhanced diagnostic strategies, including expanded microbiological testing and molecular epidemiology surveillance, could prove beneficial.
Sustained circulation of enterovirus D68 in Europe in 2023 and the continued evolution of enterovirus D68 B3-lineages associated with distinct amino acid substitutions in VP1 protein.
(Elsevier, 2025-06) Hirvonen, Aurora; Johannesen, Caroline Klint; Simmonds, Peter; Fischer, Thea K; Harvala, Heli; Benschop, Kimberley S M; ENPEN study collaborators; Instituto de Salud Carlos III; Centro de Investigación Biomédica en Red - CIBERESP (Epidemiología y Salud Pública); Unión Europea. Comisión Europea. NextGenerationEU; Helsinki University Hospital; The Slovenian Research and Innovation Agency; Rijksinstituut voor Ziekte- en Invaliditeitsverzekering; Cabrerizo, Maria; Fernandez-Garcia, Maria Dolores
Background: Enterovirus D68 (EV-D68) causes respiratory disease ranging from mild to severe and in rare cases a paralytic syndrome, called acute flaccid myelitis (AFM). Since the global EV-D68 outbreak in 2014, the virus has mainly circulated in biennial epidemic cycles with peaks detected during even years. However, following the COVID-19 pandemic, the seasonal pattern of EV-D68 has been characterized by large yearly upsurges. Here, we describe the circulation of EV-D68 in Europe in 2023 and track its genetic evolution. Study design: Data was compiled from members of the European Non-Polio Network (ENPEN). This included monthly data on the total number of EV samples tested, EV positive samples, EV-D68 positive samples and cases, and other EV positive samples detected in 2023. Information on sample types and surveillance system was recorded. Sequence data from the VP1 gene was used for phylogenetic and amino acid sequence analysis. Results: EV was detected in 13,585 out of 203,622 diagnostic samples tested (6.7 %), of which 402 (3.0 %) were determined as EV-D68, representing 386 cases. EV-D68 infections peaked in October 2023 (136/386; 35.2 %). 267/386 (69.2 %) of EV-D68 cases were captured through clinical EV surveillance, almost all of which (202/204 of positive samples with sample type information) were detected in respiratory specimens. Phylogenetic analysis performed on 99 VP1 sequences revealed a distinct B3-derived lineage with a previously undescribed residue change, D554E, in Europe. Conclusions: The study documents sustained circulation of EV-D68 in Europe in 2023, the evolution of B3-derived lineages, and appearance of previously undescribed amino acid substitutions in Europe. This stresses the need for continuous EV-D68 surveillance and harmonization of EV-D68 detection practices towards better data comparability across countries.
Extracellular vesicles derived microRNAs as non-invasive markers of liver fibrosis in chronically infected HCV patients: a pilot study.
(Elsevier, 2025-06) Cairoli, Victoria; Valle-Millares, Daniel; Ryan, Pablo; Dominguez, Lourdes; Martín-Carbonero, Luz; De Los Santos, Ignacio; De Matteo, Elena; Ameigeiras, Beatriz; De Sousa, Marcela; Briz, Veronica; Preciado, María V; Fernandez-Rodriguez, Amanda; Valva, Pamela; Instituto de Salud Carlos III; RETICS-Sida (RIS-ISCIII) (España); Centro de Investigación Biomédica en Red - CIBERINFEC (Enfermedades Infecciosas); Agencia Nacional de Promoción Científica y Tecnológica (Argentina); National Scientific and Technical Research Council (Argentina)
Extracellular vesicles (EVs) are an increasingly promising tool for liquid biopsy in liver diseases. Hepatitis C Virus (HCV) infection, alone or together with Human Immunodeficiency Virus (HIV) infection significantly impacts on the microRNA (miRNA) EVs content resembling chronic hepatitis C (CHC) progression. The objective of the study was to delve into the intricate EVs-miRNA profiles in CHC patients with different liver fibrosis stages, aiming to pinpoint non-invasive markers capable of distinguishing significant fibrosis. Plasma EV-miRNAs from 50 CHC patients (HCV+ and HCV+/HIV+) stratified in no significant (F < 2) and significant (F ≥ 2) fibrosis, were massively sequenced. General linear models (GLM) were used to identify significantly differential expressed (SDE) miRNAs according to liver fibrosis stages (F ≥ 2 and F < 2). Dysregulated biological pathways were subsequently analyzed for the following groups: i) all patients; ii) HCV+; and iii) HCV+/HIV+. Multiple-ordered logistic regression analysis was performed to develop a score to identify F ≥ 2 cases. The diagnostic potential of both the SDE miRNAs and the developed score was assessed using ROC curve analysis. With respect to all CHC patients, two SDE miRNAs (hsa-miR-122-5p and hsa-miR-92a-3p) were identified which regulate genes related to cytoskeleton organization. Regarding their diagnostic performance to discriminate F ≥ 2, both miRNAs individually demonstrated acceptable diagnostic values. However, their combined use in a new score enhanced their diagnostic performance (AUROC = 0.833). In the HCV+ subgroup, 8 SDE miRNAs (hsa-miR-122-5p, hsa-miR-320c, hsa-miR-3615, hsa-miR-320a-3p, hsa-miR-374b-5p, hsa-let-7a-3p, hsa-miR-199a-5p, hsa-miR-142-5p), which regulate macrophage activity and cell growth/death regulation, were recognized. Among them, hsa-miR-3615 displayed the highest diagnostic performance to discriminate F ≥ 2 (AUROC = 0.936). With respect to HCV+/HIV+, 18 SDE miRNAs (hsa-miR-4508, hsa-miR-122-5p, hsa-miR-451a, hsa-miR-1290, hsa-miR-1246, hsa-miR-107, hsa-miR-15b-5p, hsa-miR-194-5p, hsa-miR-22-5p, hsa-miR-20b-5p, hsa-miR-142-5p, hsa-miR-328-3p, hsa-miR-335-3p, hsa-miR-125a-5p, hsa-miR-423-3p, hsa-let-7d-3p, hsa-miR-128-3p, hsa-miR-10a-5p) were recognized that regulate RNA silencing processes. In this case, hsa-miR-423-3p and hsa-miR-128-3p showed outstanding diagnostic performances (AUROC > 0.900). Distinct EVs-miRNA profiles were identified in patients with varying liver fibrosis stages, both in the overall CHC cohort and within HCV+ and HCV+/HIV+ subgroups. These specific miRNA signatures would allow the elucidation of potential mechanisms involved in clinical evolution and identification of specific biomarkers of unfavorable progression, plausible to be used in a diagnostic panel. Furthermore, the developed score demonstrates the ability to discriminate within the CHC group those individuals with significant fibrosis regardless of their HIV infection status.
The sulfur-related metabolic status of during infection reveals cytosolic serine hydroxymethyltransferase as a promising antifungal target.
(Taylor & Francis, 2025-12) Alharthi, Reem; Sueiro-Olivares, Monica; Storer, Isabelle; Bin Shuraym, Hajer; Scott, Jennifer; Al-Shidhani, Reem; Fortune-Grant, Rachael; Bignell, Elaine; Tabernero, Lydia; Bromley, Michael; Zhao, Can; Amich, Jorge; Agencia Estatal de Investigación (España); Wellcome Trust
Sulfur metabolism is an essential aspect of fungal physiology and pathogenicity. Fungal sulfur metabolism comprises anabolic and catabolic routes that are not well conserved in mammals, therefore is considered a promising source of prospective novel antifungal targets. To gain insight into sulfur-related metabolism during infection, we used a NanoString custom nCounter-TagSet and compared the expression of 68 key metabolic genes in different murine models of invasive pulmonary aspergillosis, at 3 time-points, and under a variety of conditions. We identified a set of 15 genes that were consistently expressed at higher levels than , suggesting that they may be particularly relevant for intrapulmonary growth and thus constitute promising drug targets. Indeed, the role of 5 of the 15 genes has previously been empirically validated, supporting the likelihood that the remaining candidates are relevant. In addition, the analysis of gene expression dynamics at early (16 h), mid (24 h), and late (72 h) time-points uncovered potential disease initiation and progression factors. We further characterized one of the identified genes, encoding the cytosolic serine hydroxymethyltransferase ShmB, and demonstrated that it is an essential gene of , also required for virulence in a murine model of established pulmonary infection. We further showed that the structure of the ligand-binding pocket of the fungal enzyme differs significantly from its human counterpart, suggesting that specific inhibitors can be designed. Therefore, transcriptomics is a powerful tool for identifying genes crucial for fungal pathogenicity that may encode promising antifungal target candidates.
Sur8/Shoc2 como nuevo actor en envejecimiento y fragilidad (SENFRA-5). Data Management Plan V1.0.
(Instituto de Salud Carlos III (ISCIII). Centro Nacional de Microbiología (CNM), 2026-07-03) Fernandez-Rodriguez, Amanda; Instituto de Salud Carlos III
El presente Plan de Gestión de Datos (PGD) define las estrategias, procedimientos y responsabilidades relacionadas con la generación, almacenamiento, acceso, difusión y protección de los datos derivados del proyecto COOPERA-ISCIII. Se alinea con los criterios de evaluación del ISCIII (calidad, viabilidad e impacto) y con los principios FAIR (Findable, Accessible, Interoperable, Reusable), tal como requieren las convocatorias de la Acción Estratégica en Salud.
Bidirectional Interaction Between Liposomal Amphotericin B Pharmacokinetics and Parasite Dynamics in Patients With Post-Kala-Azar Dermal Leishmaniasis: Potential Implications for Optimal Dosing.
(Wiley, 2026-02) Chu, Wan-Yu; Singh, Om Prakash; Sundar, Shyam; Mondal, Dinesh; Pandey, Krishna; Das, Pradeep; Roseboom, Ignace C; Raja, Sheeraz; Torres Garcia, Ana Maria; Carrillo, Eugenia; Huitema, Alwin D R; Alves, Fabiana; Dorlo, Thomas P C; Dutch Research Council (Holanda); Swedish Research Council
Post-kala-azar dermal leishmaniasis (PKDL) involves a high macrophage burden in which the Leishmania parasites reside. Liposomal amphotericin B (LAmB) plays a key role in the treatment of PKDL. The mononuclear phagocyte system (MPS) is crucial in the distribution of liposomal drugs as well as the leishmaniasis pathophysiology. This study focused on characterizing the interaction between LAmB pharmacokinetics, the MPS, and parasite dynamics for optimal dosing of LAmB in PKDL. Clinical trial data from the Indian subcontinent, involving short-course LAmB administered alone or with miltefosine, were analyzed using nonlinear mixed-effects modeling. The pharmacokinetics of LAmB were best described by a two-compartment model with a saturable LAmB uptake by the MPS. The maximum MPS uptake capacity was modeled with a baseline component and an additional disease-related component relative to the parasite burden. As treatment progressed, MPS capacity decreased with declining parasite load, resulting in a median 54% increase in the systemic LAmB exposure (AUC) by the end of treatment. Simulations suggested that a similar parasite clearance could be achieved with a 50% lower total LAmB dose, supporting the potential efficacy of reduced dosing regimens. Combining LAmB and miltefosine further accelerated parasite clearance compared to LAmB alone. This study highlights the importance of understanding the bidirectional interactions between LAmB pharmacokinetics and parasite infection for interpreting systemic exposure and optimizing treatment approaches. If confirmed in clinical trials, reduced LAmB dosing strategies could enable more rational and cost-effective management of PKDL and other dermal leishmaniases.
Decoding the antiviral potential of eugenol, thymol and vanillin against human cytomegalovirus infection.
(Microbiology Society, 2026-03) Martín-Martín, Clara; Ruiz-Rico, María; Barat, José Manuel; García-Ríos, Estéfani; Pérez-Romero, Pilar; Instituto de Salud Carlos III; Agencia Estatal de Investigación (España); Generalitat Valenciana (España); University of Notre Dame
Human cytomegalovirus (HCMV) poses serious health risks, particularly for immunocompromised individuals. However, the current FDA-approved anti-HCMV drugs face challenges such as drug resistance and significant side effects, underscoring the need for alternative treatment options. Essential oil components (EOCs), including eugenol, thymol and vanillin, are recognized for their therapeutic potential. This study evaluates their antiviral effects against HCMV in epithelial (ARPE-19) and fibroblast (MRC-5) cell lines. Among the EOCs, vanillin demonstrated the highest efficacy, characterized by low toxicity and a high selectivity index in both cell types. Mechanistic differences were noted between the cell lines. In ARPE-19 cells, eugenol showed virucidal activity, inhibited viral entry and suppressed early gene expression (IE-1). Conversely, in MRC-5 cells, eugenol mainly blocked viral entry and exhibited virucidal effects. Thymol was most effective in ARPE-19 cells, where it completely suppressed IE-1 expression as a result of both inhibition of viral entry and a direct disruptive effect on IE-1 expression. In addition, thymol showed an effect on viral replication. In MRC-5 cells, thymol primarily inhibited viral entry and attachment. Vanillin exhibited dual inhibitory activity in both cell lines, blocking viral attachment and entry. In MRC-5, vanillin also appears to affect intermediate processes. Notably, combining EOCs with ganciclovir resulted in synergistic effects. The eugenol/ganciclovir combination was particularly effective in ARPE-19 cells, while thymol/ganciclovir showed enhanced efficacy in MRC-5 cells. These findings suggest that EOCs have significant potential as adjunct therapies to improve antiviral outcomes and address drug-resistant HCMV strains.
Detection and genetic characterization of Crimean-Congo hemorrhagic fever virus in ticks from western Spain (2017, 2020-2024).
(Frontiers Media, 2026-04-01) Sánchez-Mora, Patricia; Habela, Miguel A; Del Peso, Teresa; Grande Ávila, Ana Candela; García López, Ana María; Mata García Soldado, Jennifer; Tapia, María M; Molero-Sanz, Francisca; Herrero-Romero, Laura; Olmeda, A Sonia; Valcárcel, Félix; Estrada-Peña, Agustín; Negredo, Anabel; Sánchez-Seco, María Paz; Instituto de Salud Carlos III; Centro de Investigación Biomédica en Red - CIBERINFEC (Enfermedades Infecciosas); Unión Europea. Comisión Europea. NextGenerationEU
Introduction: Crimean-Congo hemorrhagic fever virus (CCHFV) was first detected in Spain in ticks collected from red deer in southwestern Cáceres. Since then, this region, established as endemic, has been the focus of several surveillance studies. However, updated data on viral circulation in this area remain limited. Materials and methods: We conducted a retrospective surveillance study to assess the presence and genetic diversity of CCHFV in ticks collected in central and southern Cáceres over multiple years (2017 and 2020-2024). A total of 3,183 ticks, grouped into 1,569 pools, were collected from wild ungulates, livestock, domestic animals and vegetation, and analyzed by two PCR methods. Positive pools were characterized by Sanger sequencing. Results: CCHFV was exclusively detected in Hyalomma lusitanicum ticks, with an overall infection rate of 1.54% (95% CI: 1.14-2.03). Most positive pools originated from wild ungulates, particularly red deer. Genetic analysis revealed the circulation of two CCHFV genotypes, predominantly genotype III. Discussion: The detection of CCHFV in ticks collected over multiple years supports the sustained circulation of the virus in southwestern Cáceres. Our findings also reinforce the key role of H. lusitanicum as the main vector maintaining the virus in wild ungulates and underscore the genetic diversity of circulating strains and the importance of using multiple molecular methods. These results emphasize the need for continuous surveillance in endemic areas to monitor viral circulation and assess animal and public health risks.
Molecular epidemiology and surveillance of imported dengue in travellers returning to Spain, 2022-2024.
(Elsevier, 2026) Sánchez-Mora, Patricia; Molero-Sanz, Francisca; Navascués, Ana; Zurita, Nelly Daniela; Zamarrón, Pilar; Lagarejos, Eduardo; Gómez, Sara; Cardona, Pere Joan; Carrasco, Blanca; López-Lomba, Marta; Escribano, Isabel; Martínez-Expósito, Óscar; Del Peso, Teresa; Herrero-Romero, Laura; Negredo, Anabel; Guillén-Calvo, Laura; Peña Gallego, Ana; Gutiérrez-López, Rafael; Labiod, Nuria; Vazquez, Ana; Sánchez-Seco, María Paz; Gutiérrez-López, Rafael; Instituto de Salud Carlos III; Centro de Investigación Biomédica en Red - CIBERINFEC (Enfermedades Infecciosas)
Introduction: Dengue is the most significant vector-borne viral disease in global morbidity and mortality, and it is considered a re-emerging disease spreading into new regions. The risk of autochthonous dengue virus transmission in Spain remains high due to the increasing number of travellers returning from endemic areas and the presence of Aedes albopictus within our territory. Conducting epidemiological and molecular studies on returning travellers from endemic areas may be crucial to discern transmission patterns and track the global spread of the virus. This study focuses on the molecular characterization of suspected imported dengue cases from 2022 to 2024. Methods: We analysed 600 samples from 539 suspected dengue-infected travellers between 2022 and 2024. All samples were tested by a quantitative RT-PCR, and PCR-positive cases were confirmed by performing a non-overlapping nested RT-PCR, going under subsequent sequencing to identify viral diversity. Results: Of the 539 suspected cases, 183 were confirmed as DENV-positive, with Cuba as the most common travel origin associated with infections. Molecular analysis of positive samples identified all four DENV serotypes and ten genotype groups, with DENV-3 genotype III as the most predominant. Additionally, among confirmed cases reported from regions with Aedes albopictus presence, 73% (94/129) occurred during the vector's active season (May-November), increasing the risk of local transmission. Conclusion: Our results highlight the importance of returning travellers as sentinels for ongoing dengue outbreaks and epidemiology in endemic regions. Additionally, these emphasize the importance of early case identification through sentinel surveillance to prevent potential autochthonous transmission.
HIV/HCV prevalence and the retention paradox in marginalized populations in Madrid (2019-2023): A large-scale cross-sectional study.
(Elsevier, 2026-06) Valencia, Jorge; Ryan, Pablo; Codina Márquez, Helena; Cuevas, Guillermo; Manzano, Samuel; Estévez, Samuel; Martinez, Isidoro; Sepulveda-Crespo, Daniel; Resino, Salvador; Gilead Sciences (Spain); Instituto de Salud Carlos III; AbbVie; Centro de Investigación Biomédica en Red - CIBERINFEC (Enfermedades Infecciosas); Unión Europea. Comisión Europea. NextGenerationEU
Objectives: Marginalized urban populations experience a high burden of HIV and HCV. We assessed the prevalence and risk factors for both infections and characterized the care cascades in a cohort of vulnerable individuals in Madrid, Spain. Methods: We conducted a cross-sectional study (2019-2023) of 4582 individuals via mobile units in high-risk hotspots, offering integrated rapid HIV/HCV testing with point-of-care HCV-RNA confirmation. Multivariable logistic regression identified independent HIV risk factors. Results: HIV prevalence was 6.3% (95% CI: 5.6-7.0). Among people with HIV (PWH), 17.0% of known cases were not receiving antiretroviral therapy; PrEP uptake was zero among HIV-negative individuals. A history of injecting drug use was the primary HIV risk factor (adjusted odds ratio[aOR] 6.6; 95% CI: 4.6-9.5), followed by age >50 years, Spanish origin, and alcohol/benzodiazepine misuse (all P < 0.05). Active HCV prevalence was 5.5% (95% CI: 4.9-6.2); 15.7% in PWH vs 4.8% in people without HIV (P < 0.001). Among confirmed cases, HCV linkage (≥95.2%) and treatment (≥88.6%) were high; however, 16.8% of all antibody-positive individuals (23.2% among PWH) missed confirmatory RNA testing. Conclusions: HIV and HCV remain prevalent, characterized by discontinuities in retention rather than diagnosis. Integrated, low-threshold strategies combining harm reduction with social support are required to address this syndemic.
Dengue nanosensors: a roadmap for clinical viability.
(Elsevier, 2026-02-26) Codina Márquez, Helena; Quero-Delgado, Marta; Gómez, Rafael; Resino, Salvador; Martinez, Isidoro; Sepulveda-Crespo, Daniel; Instituto de Salud Carlos III; Centro de Investigación Biomédica en Red - CIBERINFEC (Enfermedades Infecciosas); Unión Europea. Comisión Europea. NextGenerationEU; Centro de Investigación Biomédica en Red - CIBERBBN (Bioingeniería, Biomateriales y Nanomedicina)
Dengue nanosensors promise much but have failed in clinical application. This paradox stems from a focus on record sensitivity over real-world robustness. We propose a paradigm shift from materials discovery to product engineering, providing a translational roadmap to guide viable diagnostics from the lab bench to the patient.
Challenges in accurate HDV RNA quantification: inter-assay variability and the impact of thermal shock.
(American Society for Microbiology (ASM), 2026-02-11) Pérez-García, Felipe; Virseda-Berdices, Ana; Pita-Martínez, Carlos; Muñoz Monte, Mario; Sepulveda-Crespo, Daniel; Codina Márquez, Helena; Alonso, Roberto; Mesones, Lara; Rodrigo, Sandra; Macías, Juan; Real, Luis Miguel; Cuadros-González, Juan; Martinez, Isidoro; Resino, Salvador; Certest Biotech; Fundación para la Investigación Biomédica del Hospital Universitario Príncipe de Asturias; Instituto de Salud Carlos III; Centro de Investigación Biomédica en Red - CIBERINFEC (Enfermedades Infecciosas); Unión Europea. Comisión Europea. NextGenerationEU; European Society of Clinical Microbiology and Infectious Diseases
Quantitative RT-PCR (qRT-PCR) is essential for monitoring hepatitis delta virus (HDV) RNA, yet assays lack standardization. We aimed to evaluate the performance of three qRT-PCR assays and to assess the impact of a pre-analytical thermal shock procedure. We conducted a comparative study using 206 samples (106 with anti-HDV antibodies and 100 anti-HDV-negative as a control group), which were tested in parallel with three qRT-PCR assays: Vircell, Certest, and Altona. Performance was evaluated for inter-assay agreement (kappa index), quantitative correlation (R²), and bias (Bland-Altman). Altona detected 56 HDV-RNA positive samples, whereas Vircell and Certest detected 55 positive samples. Inter-assay agreement was perfect comparing Vircell vs Certest (agreement = 100%, к = 1.000) and almost perfect comparing Altona with both Vircell and Certest (agreement = 99.5%, к = 0.988). Quantitatively, Vircell and Certest assays showed relevant systematic biases compared to Altona, overestimating viral loads by approximately 0.24 log International Units (IU)/mL (Certest) and 0.33 log IU/mL (Vircell). The correlation with Altona was strong for Certest (R² = 0.864) and moderate for Vircell (R² = 0.793), while the correlation between Certest and Vircell was weaker (R² = 0.720). Thermal shock improved sensitivity in one case (Certest vs Vircell) but increased quantitative variability, worsening the inter-assay correlation (R² = 0.684). In conclusion, all three assays were highly concordant for the qualitative diagnosis of HDV infection, but their quantitative biases prevent their interchangeable use for treatment monitoring. Thermal shock is not recommended for routine monitoring, as a significant compromise in quantitative accuracy and precision outweighs any potential gains in sensitivity.IMPORTANCEThis study evaluates three hepatitis delta virus (HDV) RNA quantitative real-time PCR (qRT-PCR) assays, crucial for managing the HDV infection, particularly in the setting of new therapies like Bulevirtide, where assessing viral load reduction and accurate monitoring is paramount. We reveal significant quantitative biases among widely used assays, precluding their interchangeable use and risking misinterpretation of treatment response. Furthermore, our systematic assessment of the thermal shock pre-analytical procedure highlights its detrimental impact on quantitative precision, despite modest sensitivity gains. This work provides essential evidence for clinicians and laboratories, guiding assay selection and standardization efforts to optimize HDV diagnosis and patient monitoring.
Sex and age-related differences in humoral and cellular responses to the recombinant zoster vaccine in people living with HIV.
(Elsevier, 2025-11) Díez, Cristina; Pita-Martínez, Carlos; Quero-Delgado, Marta; Gómez-Tórtola, Aurora; Fanciulli, Chiara; Marchan, Ana; Aldámiz-Echevarría, Teresa; Sepulveda-Crespo, Daniel; Tejerina, Francisco; Pérez-Latorre, Leire; Gutiérrez, Isabel; López, Juan Carlos; Martinez, Isidoro; Resino, Salvador; Agencia Estatal de Investigación (España); Centro de Investigación Biomédica en Red - CIBERINFEC (Enfermedades Infecciosas); Instituto de Salud Carlos III; Unión Europea. Comisión Europea. NextGenerationEU
Background: Data on the immunogenicity of the recombinant zoster vaccine (RZV) in people living with HIV (PLWH) are limited, despite their high risk for herpes zoster. This study aimed to characterize the humoral (Varicella-zoster virus (VZV)-specific IgG) and cellular (VZV-specific IFN-γ/IL-2 T-cell) immune responses to RZV in PLWH on suppressive antiretroviral therapy (ART), stratified by sex and age. Methods: This prospective study enrolled 207 PLWH on suppressive ART who received two doses of RZV. VZV-specific IgG antibody titers were quantified by in-house ELISA at baseline and post-vaccination and T-cell responses using FluoroSpot assays post-vaccination. We estimated geometric mean titers (GMTs), geometric mean fold rises (GMFRs), and adjusted arithmetic mean ratios (aAMRs) using generalized linear models. Results: RZV vaccination induced a significant humoral response, with an overall GMFR of 16.7 and a 71 % positive response rate (≥4-fold rise in IgG titers). Females < 60 years old exhibited a markedly higher GMFR (43.8) than all other subgroups (p < 0.05). The vaccine also induced robust VZV-specific T-cell responses, with GMTs being comparable across all groups. Notably, a strong positive association was found between humoral and T-cell responses, particularly with IL-2-secreting cells (aAMR=1.6, p < 0.001). This association was most pronounced in participants < 60 years, especially females, and was attenuated in older individuals. Conclusions: RZV vaccination effectively induces both humoral and T-cell immunity in PLWH. However, host factors, such as sex and age, critically modulate immunogenicity. Females < 60 years showed a superior humoral response and the strongest association between immune compartments, highlighting important variability in vaccine responsiveness.
Identification and functional insights into new phage tail-like bacteriocins targeting as new antimicrobials.
(American Society for Microbiology (ASM), 2026-03-24) Ibarguren-Quiles, Clara; Blasco, Lucía; López-Causape, Carla; Bleriot, Inés; Fernández-García, Laura; Arman, Lucia; Barrio-Pujante, Antonio; Ortiz-Cartagena, Concha; Aracil, Belen; Menéndez-Rodriguez, Olaya; Mariñas-Pardo, Luis; Cantón, Rafael; Oliver, Antonio; Tomás, María; Instituto de Salud Carlos III; Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF); Centro de Investigación Biomédica en Red - CIBERINFEC (Enfermedades Infecciosas); Ministerio de Ciencia, Innovación y Universidades (España); Unión Europea. Comisión Europea. NextGenerationEU; Xunta de Galicia (España)
The current health crisis caused by multidrug-resistant (MDR) pathogens is one of the health problems of most concern globally. Infections caused by these pathogens, such as , lead to high rates of complications, particularly in compromised patients such as cystic fibrosis (CF) patients. The need to counteract and minimize the forecast future impact has led to the rescue of phage therapy. The use of bacteriophages has important advantages, including highly specific targeting, self-amplification at the infection site, minimal disruption of the microbiome, safety, and biocompatibility. However, the capacity of bacteria to escape these entities results in a form of resistance that compromises the effectiveness of the therapy. This involves the search for potential alternatives, such as the phage tail-like bacteriocins (PTLBs), also named as tailocins. These high-molecular-weight particles resemble the tail structure of bacteriophages and are characterized by the absence of genetic material, avoiding the development of resistance, one of the major handicaps associated with phage therapy. In this study, we detected 34 different PTLBs in 75 genomes, with different serotypes and sequence types, 11 of which were characterized as novel F-type PTLB subtypes (F13-F24). Furthermore, we report that four selected PTLBs (R1, F15, F19, and R3-F24) can deal with bacterial infection, with the R1 and the F15 PTLBs being the most efficient in clearing infection , yielding a survival rate of more than 75% in the larvae model. This reaffirms the potential of PTLBs to control infections, which can cause chronic infections in some patients, such as people with CF, due to its strong impact as a MDR bacterium.IMPORTANCEThe 75 genomes from people with cystic fibrosis in the study collection included at least one phage tail-like bacteriocins (PTLB) cluster. From the 34 different PTLBs detected in the study collection, 7 were R-type, 10 were complex (R- and F-type encoded), and 14 were F-type PTLBs. Eleven new F-type PTLBs were described in the collection under study. An association between the O-antigen present on the surface of the isolate and the encoded PTLB subtype was detected. The R1 and F15 PTLB subtypes display high antimicrobial activity both in vitro and in vivo (Galleria mellonella).
Advances in Nanotechnology-Enabled Optical Biosensors for Dengue Virus Detection: A Systematic Review.
(Wiley, 2025-07-29) Quero-Delgado, Marta; Codina Márquez, Helena; Gómez, Rafael; Terán, Francisco José; Muñoz-Fernández, M A; Jiménez, José Luis; Resino, Salvador; Sepulveda-Crespo, Daniel; Martinez, Isidoro; Instituto de Salud Carlos III; Centro de Investigación Biomédica en Red - CIBERINFEC (Enfermedades Infecciosas); Centro de Investigación Biomédica en Red - CIBERBBN (Bioingeniería, Biomateriales y Nanomedicina); Comunidad de Madrid (España)
The dramatic surge in dengue cases in early 2024, endangering half the global population, urgently necessitates faster diagnostic methods. Nanotechnology-enabled optical biosensors offer a promising avenue, leveraging nanomaterial properties for highly sensitive detection of dengue virus (DENV), potentially surpassing conventional techniques in terms of simplicity, speed, and cost-effectiveness. This systematic review analyzes recent advancements in these biosensors for DENV diagnosis. Following PRISMA 2020 guidelines, we systematically searched PubMed, Embase, Scopus, Web of Science, and Cochrane Library databases (2010-June 1, 2025; OSF: https://osf.io/3gmey/). The methodological quality of the 98 included studies was assessed using a modified CASP checklist. Diverse nanotechnology-based optical biosensors were identified. SPR (36.7%) was the most common transducer, followed by fluorescence (24.5%), colorimetry (15.3%), and SERS (8.2%). Gold-based nanomaterials (35.7%) were most frequently employed, with silver nanomaterials (15.3%), quantum dots (15.3%), and graphene-based materials (15.3%) also showing promise. DENV NS1 protein was the primary target analyte (21.4%). Importantly, almost half of the studies (44.9%) used clinically relevant human samples. While many optical biosensors show promise, challenges hinder their demonstration of the true potential for point-of-care use in their current format; however, they offer high specificity and faster results, laying a strong foundation for cost-effective clinical diagnostics. Nanotechnology-driven optical biosensors offer a transformative approach for DENV detection. Advances in computational design and green synthesis of novel nanomaterials are key to addressing stability and field-deployment challenges. These innovations are crucial for developing robust, sensitive, and user-friendly tools to manage dengue and improve patient outcomes globally.
From Management to Cure: The Shifting Paradigm in HIV and Chronic Viral Hepatitis.
(Multidisciplinary Digital Publishing Institute (MDPI), 2025-07-11) Sepulveda-Crespo, Daniel; Resino, Salvador
The management of human immunodeficiency virus (HIV) and chronic viral hepatitis (HBV, HCV, and HDV) infections continues to pose a significant global health challenge [...].
Prevalence and risk factors of active hepatitis C infection among at-risk migrant populations in Madrid, Spain, 2019 to 2023.
(European Centre for Disease Prevention and Control (ECDC), 2025-07) Ryan, Pablo; Valencia, Jorge; Pérez-García, Felipe; Quero-Delgado, Marta; Cuevas, Guillermo; Manzano, Samuel; Estévez, Samuel; Martinez, Isidoro; Sepulveda-Crespo, Daniel; Resino, Salvador; Gilead Sciences (Spain); Instituto de Salud Carlos III; AbbVie; Asociación Española para el Estudio del Hígado; Centro de Investigación Biomédica en Red - CIBERINFEC (Enfermedades Infecciosas)
Background: Hepatitis C virus (HCV) microelimination among at-risk migrants supports global elimination goals.AIMTo evaluate risk factors, prevalence and trends of active HCV infection among at-risk migrants screened for HCV in Madrid from 2019-23. Methods: At-risk migrants (born outside Spain, living in country < 10 years regardless of legal status), were screened for HCV via mobile units with rapid antibody testing, and confirmed by RNA testing. Recruitment of this convenience sample focused on migrant centres, shelters, harm reduction centres and social service sites. Primary outcome was active HCV prevalence. Risk factors analysed included origin, alcohol use, no stable income, drug use and sexual behaviour. Data were analysed using general linear models with negative binomial distribution and p values adjusted for multiple comparisons (q values). Results: TSAmong 2,288 migrants, 6.5% (149/2,288) had anti-HCV antibodies, 47.0% (70/149) of whom tested positive for HCV-RNA; 81.4% (57/70) began antiviral therapy. Overall prevalence of active HCV infection was 3.1% (70/2,288). Injection drug use (non-active vs never used (aIRR: 7.3; 95% CI: 2.7-12.7) and active (aIRR: 14.7; 95% CI: 6.7-32.1)), European origin (vs non-European; aIRR: 5.8; 95% CI: 2.7-12.7) and alcohol misuse (vs no misuse; aIRR: 1.8; 95% CI: 1.1-2.9) were main risk factors. Prevalence showed no significant change during 2019-23 in the overall population and across risk groups. Conclusion: At-risk migrants screened in Madrid had a high prevalence of active HCV infection. This is higher than reported estimates for the general Spanish population and supports the need to enhance targeted HCV prevention, screening and treatment strategies among migrant populations.
Persistent Low Anti-HIV Neutralizing Antibody Titers in HIV/HCV Coinfection Despite HCV Cure: A 5-Year Longitudinal Analysis.
(Multidisciplinary Digital Publishing Institute (MDPI), 2025-05-19) Sepulveda-Crespo, Daniel; Sánchez-Merino, Víctor; Amigot-Sánchez, Rafael; Rubio-Pérez, Almudena; Díez, Cristina; Hontañón, Víctor; Berenguer, Juan; González-García, Juan; García, Felipe; Martinez, Isidoro; Yuste, Eloísa; Resino, Salvador; Instituto de Salud Carlos III; Centro de Investigación Biomédica en Red - CIBERINFEC (Enfermedades Infecciosas)
Background: Anti-HIV neutralizing antibodies (anti-HIV-nAbs) play a critical role in the immune defense against HIV by preventing viral entry and limiting replication. This study longitudinally evaluated the titers and variability of anti-HIV-nAbs in individuals coinfected with HIV and HCV. Samples were collected at three time points: before starting HCV treatment, one year after completion, and five years post-treatment. Methods: A retrospective analysis was conducted on 71 HIV/HCV-coinfected patients who achieved a sustained virologic response following antiviral therapy for HCV. A control group of 41 HIV-monoinfected individuals was also included. Anti-HIV-nAb titers were evaluated by HIV neutralization assays using a panel of six recombinant HIV viruses representing multiple genetic subtypes. Generalized Linear Mixed Models and Generalized Linear Models were used for statistical analysis. p-values were adjusted using the Benjamini-Hochberg procedure (q-value). Results: HIV-neutralizing antibody responses in HIV/HCV-coinfected individuals remained stable over five years following HCV therapy without significant changes (q-value > 0.05). The mean neutralization scores remained stable, with baseline scores of 6.1 (95% CI: 5.4-6.7), 6.2 (95% CI: 5.5-6.8) at one year post-HCV therapy, and 6.0 (95% CI: 5.3-6.7) at five years post-HCV therapy. HIV/HCV-coinfected individuals consistently showed lower neutralization scores compared to the control group throughout the follow-up (q-value < 0.05). Regression analyses adjusted for age, gender, nadir CD4+, and baseline CD4+ counts confirmed that the observed differences between HIV-monoinfected and HIV/HCV-coinfected individuals persisted (q-value < 0.05) at both the baseline and after HCV therapy completion. Conclusions: Successful HCV eradication in HIV/HCV-coinfected individuals did not normalize anti-HIV-nAb titers, which remained consistently lower than those in HIV-monoinfected controls over five years.
Antibody Response Against SARS-CoV-2 Spike Protein in People with HIV After COVID-19 Vaccination.
(Multidisciplinary Digital Publishing Institute (MDPI), 2025-04-29) Muñoz-Gómez, María José; Ryan, Pablo; Quero-Delgado, Marta; Martin-Vicente, Maria; Cuevas, Guillermo; Valencia, Jorge; Jiménez, Eva; Blanca-López, Natalia; Manzano, Samuel; Lazo, Juan Ignacio; Mas, Vicente; Vázquez, Mónica; Sepulveda-Crespo, Daniel; Torres-Macho, Juan; Martinez, Isidoro; Resino, Salvador; Instituto de Salud Carlos III; Centro de Investigación Biomédica en Red - CIBERINFEC (Enfermedades Infecciosas)
People with HIV (PWH) often have a suboptimal response to vaccines, raising concerns regarding the efficacy of coronavirus disease 2019 (COVID-19) vaccines in this population. We aimed to evaluate the humoral immune response to the B.1 lineage and Omicron variant in PWH on antiretroviral therapy (ART) following COVID-19 vaccination. : We conducted a prospective study of 19 PWH on ART who received a two-dose series of the COVID-19 mRNA vaccine and a booster six months later. Participants without HIV infection (n = 25) were included as a healthy control (HC) group. The humoral response to the COVID-19 vaccine (anti-SARS-CoV-2 S IgG levels and ability to block ACE2-S interaction) against both the original B.1 lineage and the Omicron variant was assessed using immunoassays. : The humoral response in PWH was very strong (geometric mean fold rise, GMFR > 8) after the second dose and strong (GMFR > 4) after the booster dose for both the B.1 lineage and the Omicron variant. We found comparable humoral responses to the B.1 lineage and Omicron variant between PWH and HC groups after the second and booster doses (q-value > 0.05). The COVID-19 vaccine generated a significantly weaker humoral response against the Omicron variant compared to the B.1 lineage in both groups (q-value < 0.05). However, this response improved after the booster dose, although it remained weaker in PWH. : PWH showed a strong humoral response to the COVID-19 vaccine against B.1 and Omicron, though the Omicron response was weaker than B.1. Booster doses in PWH improved the Omicron response, but it stayed lower than B.1. Findings confirm vaccine effectiveness in PWH, stressing the critical role of boosters and potential need for updated vaccines for variants like Omicron.

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