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Search Results (227)

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Keywords = respiratory pathogens diagnosis

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12 pages, 1742 KiB  
Article
Detection of Microorganisms Causing Human Respiratory Infection Using One-Tube Multiplex PCR
by Isabela L. Lima, Adriana F. Neves, Robson J. Oliveira-Júnior, Lorrayne C. M. G. Honório, Vitória O. Arruda, Juliana A. São Julião, Luiz Ricardo Goulart Filho and Vivian Alonso-Goulart
Infect. Dis. Rep. 2025, 17(4), 93; https://doi.org/10.3390/idr17040093 - 4 Aug 2025
Viewed by 52
Abstract
Background/Objectives: Due to the significant overlap in symptoms between COVID-19 and other respiratory infections, a multiplex PCR-based platform was developed to simultaneously detect 22 respiratory pathogens. Target sequences were retrieved from the GenBank database and aligned using Clustal Omega 2.1 to identify conserved [...] Read more.
Background/Objectives: Due to the significant overlap in symptoms between COVID-19 and other respiratory infections, a multiplex PCR-based platform was developed to simultaneously detect 22 respiratory pathogens. Target sequences were retrieved from the GenBank database and aligned using Clustal Omega 2.1 to identify conserved regions prioritized for primer design. Primers were designed using Primer Express® 3.0.1 and evaluated in Primer Explorer to ensure specificity and minimize secondary structures. A multiplex strategy organized primers into three groups, each labeled with distinct fluorophores (FAM, VIC, or NED), allowing for detection by conventional PCR or capillary electrophoresis (CE). Methods: After reverse transcription for RNA targets, amplification was performed in a single-tube reaction. A total of 340 clinical samples—nasopharyngeal and saliva swabs—were collected from patients, during the COVID-19 pandemic period. The automated analysis of electropherograms enabled precise pathogen identification. Results: Of the samples analyzed, 57.1% tested negative for all pathogens. SARS-CoV-2 was the most frequently detected pathogen (29%), followed by enterovirus (6.5%). Positive results were detected in both nasopharyngeal and saliva swabs, with SARS-CoV-2 predominating in saliva samples. Conclusion: This single-tube multiplex PCR-CE assay represents a cost-effective and robust approach for comprehensive respiratory pathogen detection. It enables rapid and simultaneous diagnosis, facilitating targeted treatment strategies and improved patient outcomes. Full article
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8 pages, 9195 KiB  
Case Report
Fatal Case of Viral Pneumonia Associated with Metapneumovirus Infection in a Patient with a Burdened Medical History
by Parandzem Khachatryan, Naira Karalyan, Hasmik Petunts, Sona Hakobyan, Hranush Avagyan, Zarine Ter-Pogossyan and Zaven Karalyan
Microorganisms 2025, 13(8), 1790; https://doi.org/10.3390/microorganisms13081790 - 31 Jul 2025
Viewed by 220
Abstract
Background: Human metapneumovirus (hMPV) is a respiratory pathogen that causes illness ranging from mild upper respiratory tract infections to severe pneumonia, particularly in individuals with comorbidities. Fatal cases of hMPV-induced hemorrhagic pneumonia are rare and likely under-reported. Diagnosis is often delayed due to [...] Read more.
Background: Human metapneumovirus (hMPV) is a respiratory pathogen that causes illness ranging from mild upper respiratory tract infections to severe pneumonia, particularly in individuals with comorbidities. Fatal cases of hMPV-induced hemorrhagic pneumonia are rare and likely under-reported. Diagnosis is often delayed due to overlapping symptoms with other respiratory viruses and the rapid progression of the disease. Case presentation: We report the case of a 55-year-old man with a complex medical history, including liver cirrhosis and diabetes mellitus, who developed acute viral pneumonia. Initial symptoms appeared three days before a sudden clinical deterioration marked by shortness of breath, hemoptysis, and respiratory failure. A nasopharyngeal swab taken on the third day of illness tested positive for hMPV by qRT-PCR. The patient died the following day. Postmortem molecular testing confirmed hMPV in lung tissue and alveolar contents. Autopsy revealed bilateral hemorrhagic pneumonia with regional lymphadenopathy. Histopathological examination showed alveolar hemorrhage, multinucleated cells, neutrophilic infiltration, activated autophagy in macrophages, and numerous cytoplasmic eosinophilic viral inclusions. Conclusions: This is the first documented case of fatal hMPV pneumonia in Armenia. It highlights the potential severity of hMPV in adults with chronic health conditions and emphasizes the need for timely molecular diagnostics. Postmortem identification of characteristic viral inclusions may serve as a cost-effective histopathological marker of hMPV-associated lung pathology. Full article
(This article belongs to the Section Virology)
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16 pages, 720 KiB  
Article
Demographic and Clinical Profile of Patients with Osteogenesis Imperfecta Hospitalized Due to Coronavirus Disease (COVID)-19: A Case Series of 13 Patients from Brazil
by Luana Lury Morikawa, Luiz Felipe Azevedo Marques, Adriele Evelyn Ferreira Silva, Patrícia Teixeira Costa, Lucas Silva Mello, Andrea de Melo Alexandre Fraga and Fernando Augusto Lima Marson
Healthcare 2025, 13(15), 1779; https://doi.org/10.3390/healthcare13151779 - 23 Jul 2025
Viewed by 265
Abstract
Background: Osteogenesis imperfecta (OI) is a rare genetic connective tissue disorder characterized by bone fragility, most often caused by pathogenic variants in type I collagen genes. In this context, we aimed to describe the clinical and epidemiological characteristics of patients with OI who [...] Read more.
Background: Osteogenesis imperfecta (OI) is a rare genetic connective tissue disorder characterized by bone fragility, most often caused by pathogenic variants in type I collagen genes. In this context, we aimed to describe the clinical and epidemiological characteristics of patients with OI who were hospitalized for coronavirus disease (COVID)-19 in Brazil between 2020 and 2024. Methods: We conducted a retrospective descriptive analysis using data from the Brazilian Unified Health System (SUS, which stands for the Portuguese Sistema Único de Saúde) through the Open-Data-SUS platform. Patients with a confirmed diagnosis of OI and hospitalization due to COVID-19 were included. Descriptive statistical analysis was performed to evaluate demographic, clinical, and outcome-related variables. We included all hospitalized COVID-19 cases with a confirmed diagnosis of OI between 2020 and 2024. Results: Thirteen hospitalized patients with OI and COVID-19 were identified. Most were adults (9; 69.2%), male (7; 53.8%), self-identified as White (9; 69.2%), and all were residents of urban areas (13; 100.0%). The most frequent symptoms were fever (10; 76.9%), cough (9; 69.2%), oxygen desaturation (9; 69.2%), dyspnea (8; 61.5%), and respiratory distress (7; 53.8%). Two patients had heart disease, one had chronic lung disease, and one was obese. As for vaccination status, five patients (38.5%) had been vaccinated against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Four patients (30.8%) required admission to an intensive care unit (ICU), and six (46.2%) required noninvasive ventilatory support. Among those admitted to the ICU, only two required invasive mechanical ventilation. The clinical outcome was death in two cases (15.4%). Both patients were male, White, and had not been vaccinated against SARS-CoV-2. One was 47 years old, was not admitted to the ICU, but required noninvasive ventilation. Despite the underlying condition most patients had favorable outcomes, consistent with an international report. Conclusions: This is the first report to describe the clinical and epidemiological profile of patients with OI hospitalized for COVID-19 in Brazil, providing initial insights into how a rare bone disorder intersects with an acute respiratory infection. The generally favorable outcomes observed—despite the underlying skeletal fragility—suggest that individuals with OI are not necessarily at disproportionate risk of severe COVID-19, particularly when appropriately monitored. The occurrence of deaths only among unvaccinated patients underscores the critical role of SARS-CoV-2 vaccination in this population. Although pharmacological treatment data were unavailable, the potential protective effects of bisphosphonates and vitamin D merit further exploration. These findings support the need for early preventive strategies, systematic vaccination efforts, and dedicated clinical protocols for rare disease populations during infectious disease outbreaks. Full article
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11 pages, 449 KiB  
Article
Implementation of a Laboratory-Developed Test for the Diagnosis of Mycoplasma pneumoniae Using a High-Throughput Approach
by Valeria Conciatori, Sarah Di Sopra, Elisa Franchin, Ioannis Bekas, Giuseppe Di Pietra, Ignazio Castagliuolo, Cristiano Salata and Claudia Del Vecchio
Pathogens 2025, 14(7), 692; https://doi.org/10.3390/pathogens14070692 - 14 Jul 2025
Viewed by 366
Abstract
Mycoplasma pneumoniae is a significant causative agent of atypical pneumonia in both children and adults. Timely and accurate diagnosis is crucial for appropriate patient management. Conventional methods for detecting M. pneumoniae, such as culture and serology, exhibit several limitations regarding sensitivity, specificity, [...] Read more.
Mycoplasma pneumoniae is a significant causative agent of atypical pneumonia in both children and adults. Timely and accurate diagnosis is crucial for appropriate patient management. Conventional methods for detecting M. pneumoniae, such as culture and serology, exhibit several limitations regarding sensitivity, specificity, and turnaround time. In contrast, real-time PCR is considered the most reliable, rapid, and sensitive technique for the diagnosis of M. pneumoniae infection. In this study, we adapted and validated an in-house real-time PCR assay for use on the fully automated Panther Fusion® System. The validation process included two artificial samples, five external quality controls, and sixty-two patient samples. We evaluated the performance in terms of precision, sensitivity, linearity, and analytical sensitivity, comparing it to the original in-house assay. The Panther Fusion® System demonstrated a broad dynamic range (16–1.6 × 107 copies/reaction), a robust correlation (94%) with the in-house assay, and comparable sensitivity (46 copies/mL vs. 25 copies/mL). The concordance between the in-house real-time PCR and the Panther Fusion® System was 100% for both clinical samples and external quality controls. The adaptation of the test to the Panther Fusion® System enabled the inclusion of M. pneumoniae among the pathogens monitored for respiratory infection surveillance. Throughout 2024, we analyzed 2567 samples, with a peak positivity rate of 38% observed in August. These findings underscore the significance of employing the M. pneumoniae diagnostic assay on the Panther Fusion® System which proves valuable for the detection of M. pneumoniae infections. This platform offers the advantages of increased automation and greater throughput potential compared to other platforms, enhancing the efficiency of respiratory pathogen detection in clinical settings. Full article
(This article belongs to the Section Bacterial Pathogens)
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19 pages, 3179 KiB  
Article
Development of a Multiplex Real-Time PCR Assay for the Detection of Eight Pathogens Associated with Bovine Respiratory Disease Complex from Clinical Samples
by Fuxing Hao, Chunhao Tao, Ruilong Xiao, Ying Huang, Weifeng Yuan, Zhen Wang and Hong Jia
Microorganisms 2025, 13(7), 1629; https://doi.org/10.3390/microorganisms13071629 - 10 Jul 2025
Viewed by 365
Abstract
Bovine respiratory disease complex (BRDC) is one of the primary causes of morbidity, mortality, and economic loss in cattle worldwide. Accurate and rapid identification of causative pathogenic agents is essential for effective disease management and control. In this study, a novel multiplex fluorescence-based [...] Read more.
Bovine respiratory disease complex (BRDC) is one of the primary causes of morbidity, mortality, and economic loss in cattle worldwide. Accurate and rapid identification of causative pathogenic agents is essential for effective disease management and control. In this study, a novel multiplex fluorescence-based quantitative polymerase chain reaction (qPCR) assay was developed for the simultaneous detection of eight major pathogens associated with BRDC. The targeted pathogens included the following: bovine viral diarrhea virus (BVDV), bovine parainfluenza virus type 3 (BPIV3), bovine respiratory syncytial virus (BRSV), bovine coronavirus (BcoV), Mycoplasma bovis (M.bovis), Pasteurella multocida (PM), Mannheimia haemolytica (MH), and infectious bovine rhinotracheitis virus (IBRV). The assay was rigorously optimized to ensure high specificity with no cross-reactivity among targets. The limit of detection (LOD) was determined to be as low as 5 copies per reaction for all target pathogens. The coefficient of variation (CVs) for both intra-assay and inter-assay measurements were consistently below 2%, demonstrating excellent reproducibility. To validate the clinical utility of the assay, a total of 1012 field samples were tested, including 504 nasal swabs from Farm A and 508 from Farm B in Jiangsu Province. BVDV, BcoV, PM, and MH were detected from Farm A, with a BVDV-positive rate of 21.63% (109/504), BcoV-positive rate of 26.79% (135/504), PM-positive rate of 28.77% (145/504), and MH-positive rate of 15.08% (76/504). Also, BcoV, PM, MH, and IBRV were detected from Farm B, with a BcoV-positive rate of 2.36% (12/508), PM-positive rate of 1.38% (7/508), MH-positive rate of 14.76% (75/508), and IBRV-positive rate of 5.51% (28/508). Notably, a significant proportion of samples showed evidence of mixed infections, underscoring the complexity of BRDC etiology and the importance of a multiplex diagnostic approach. In conclusion, the developed multiplex qPCR assay provides a reliable, rapid, and cost-effective tool for simultaneous detection of multiple BRDC-associated pathogens, which will hold great promise for enhancing disease surveillance, early diagnosis, and targeted intervention strategies, ultimately contributing to improved BRDC management and cattle health outcomes. Full article
(This article belongs to the Special Issue Animal Viral Infectious Diseases)
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12 pages, 3967 KiB  
Article
Development and Application of a Multiplex Real-Time TaqMan qPCR Assay for the Simultaneous Detection of African Swine Fever Virus, Classical Swine Fever Virus, Porcine Reproductive and Respiratory Syndrome Virus, Pseudorabies Virus, and Porcine Circovirus Type 2
by Dongdong Yin, Shuangshuang Xu, Yayun Liu, Hao Guo, Mengdie Lan, Lei Yin, Jieru Wang, Yin Dai, Xuehuai Shen, Kai Zhan and Xiaocheng Pan
Microorganisms 2025, 13(7), 1573; https://doi.org/10.3390/microorganisms13071573 - 3 Jul 2025
Viewed by 420
Abstract
Since its emergence in China in 2018, African swine fever virus (ASFV) has posed a severe threat to the pig farming industry due to its high transmissibility and mortality rate. The clinical signs of ASFV infection often overlap with those caused by other [...] Read more.
Since its emergence in China in 2018, African swine fever virus (ASFV) has posed a severe threat to the pig farming industry due to its high transmissibility and mortality rate. The clinical signs of ASFV infection often overlap with those caused by other swine viruses such as classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), and porcine circovirus type 2 (PCV2), making timely and precise diagnosis a considerable challenge. To address this, we established a TaqMan-based multiplex real-time quantitative PCR (qPCR) assay capable of simultaneously detecting ASFV, CSFV, PRRSV, PRV, and PCV2. Specific primer-probe sets were developed targeting conserved genomic regions: the ASFV P72 gene, CSFV 5’UTR region, PRRSV ORF6, PCV2 cap gene, and PRV gB gene. After thorough optimization, the assay demonstrated robust analytical performance, exhibiting strong target specificity with no cross-detection of non-target pathogens. The detection threshold was determined to be 10 copies/μL per virus, indicating high assay sensitivity. Repeatability analysis revealed low variability, with intra- and inter-assay coefficient of variation values remaining below 2.3%. When applied to 95 clinical samples, the multiplex assay yielded results that were fully consistent with those obtained using commercially available singleplex qPCR kits. In conclusion, the multiplex TaqMan qPCR method developed in this study is characterized by high specificity, sensitivity, and reproducibility. It provides a reliable and efficient diagnostic tool for the simultaneous detection and differential diagnosis of ASFV and other clinically similar viral infections in swine, thereby offering robust technical support for swine disease surveillance and control. Full article
(This article belongs to the Special Issue Viral Infection on Swine: Pathogenesis, Diagnosis and Control)
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16 pages, 904 KiB  
Review
Point-of-Care Diagnostic Testing for Emerging and Existing Poultry Viral Respiratory Pathogens Using Loop-Mediated Isothermal Amplification
by Ben Enyetornye, Aurelle Yondo and Binu T. Velayudhan
Pathogens 2025, 14(7), 657; https://doi.org/10.3390/pathogens14070657 - 2 Jul 2025
Viewed by 335
Abstract
Accurate, rapid and inexpensive diagnosis of poultry respiratory pathogens remains a challenge, especially in many developing countries. Meanwhile, poultry respiratory pathogens are a major threat to poultry production worldwide, accounting for billions of dollars in economic loss to the sector. Early and accurate [...] Read more.
Accurate, rapid and inexpensive diagnosis of poultry respiratory pathogens remains a challenge, especially in many developing countries. Meanwhile, poultry respiratory pathogens are a major threat to poultry production worldwide, accounting for billions of dollars in economic loss to the sector. Early and accurate diagnosis of these diseases is critical for economic poultry production. Molecular diagnostic methods, including PCR-based techniques, have been developed and used to fill this gap, but unfortunately, these techniques require skilled technicians, relatively costly equipment and reagents and can only be performed in a laboratory setting. This warrants the development of other diagnostic tools, which can be used in the field even by unskilled personnel. In this review, we discussed the genesis, challenges, advances and prospects of loop-mediated isothermal amplification (LAMP) for the detection of poultry respiratory pathogens at the flock side, especially in resource-constrained countries. We highlighted the application of LAMP in routine poultry disease surveillance and early outbreak detection, underscoring its value as a transformative diagnostic tool in poultry production. The development and use of a point-of-care (POC) LAMP assay that can be used to screen for these poultry respiratory pathogens simultaneously enhance disease surveillance and diagnosis. Full article
(This article belongs to the Special Issue Diagnostics of Emerging and Re-Emerging Pathogens)
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25 pages, 1270 KiB  
Review
Biofilm Formation of Pseudomonas aeruginosa in Cystic Fibrosis: Mechanisms of Persistence, Adaptation, and Pathogenesis
by Dayana Borisova, Tsvetelina Paunova-Krasteva, Tanya Strateva and Stoyanka Stoitsova
Microorganisms 2025, 13(7), 1527; https://doi.org/10.3390/microorganisms13071527 - 30 Jun 2025
Viewed by 838
Abstract
Cystic fibrosis (CF) is a life-limiting autosomal recessive disorder affecting a large number of individuals in Europe. The disease arises from mutations in the CFTR gene encoding the cystic fibrosis transmembrane conductance regulator, a chloride ion channel crucial for maintaining epithelial ion and [...] Read more.
Cystic fibrosis (CF) is a life-limiting autosomal recessive disorder affecting a large number of individuals in Europe. The disease arises from mutations in the CFTR gene encoding the cystic fibrosis transmembrane conductance regulator, a chloride ion channel crucial for maintaining epithelial ion and fluid homeostasis. Dysfunctional CFTR disrupts mucociliary clearance, particularly in the respiratory tract, resulting in persistent bacterial colonization, chronic inflammation, and progressive pulmonary damage—ultimately leading to respiratory failure, the principal cause of mortality in CF patients. Early diagnosis and advances in therapy have substantially improved both survival and quality of life. A hallmark of CF pathology is the establishment of polymicrobial infections within the thickened airway mucus. Pseudomonas aeruginosa is the dominant pathogen in chronic CF lung infections and demonstrates a remarkable capacity for adaptation via biofilm formation, metabolic reprogramming, and immune evasion. Biofilms confer increased tolerance to antimicrobial agents and facilitate long-term persistence in hypoxic, nutrient-limited microenvironments. P. aeruginosa exhibits a wide range of virulence factors, including exotoxins (e.g., ExoU, ExoS), pigments (pyoverdine, pyochelin), and motility structures (flagella and pili), which contribute to tissue invasion, immune modulation, and host damage. During chronic colonization, P. aeruginosa undergoes significant genotypic and phenotypic changes, such as mucoid conversion, downregulation of acute virulence pathways, and emergence of hypermutator phenotypes that facilitate rapid adaptation. Persistent cells, a specialized subpopulation characterized by metabolic dormancy and antibiotic tolerance, further complicate eradication efforts. The dynamic interplay between host environment and microbial evolution underlies the heterogeneity of CF lung infections and presents significant challenges for treatment. Elucidating the molecular mechanisms driving persistence, hypermutability, and biofilm resilience is critical for the development of effective therapeutic strategies targeting chronic P. aeruginosa infections in CF. Full article
(This article belongs to the Section Biofilm)
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10 pages, 778 KiB  
Case Report
A Rare Case of Exophiala Dermatitidis Isolation in a Patient with Non-Cystic Fibrosis Bronchiectasis: Colonization or True Infection?
by Francesco Rocco Bertuccio, Nicola Baio, Simone Montini, Valentina Ferroni, Vittorio Chino, Lucrezia Pisanu, Marianna Russo, Ilaria Giana, Elisabetta Gallo, Lorenzo Arlando, Klodjana Mucaj, Mitela Tafa, Maria Arminio, Emanuela De Stefano, Alessandro Cascina, Angelo Guido Corsico, Giulia Maria Stella and Valentina Conio
Diagnostics 2025, 15(13), 1661; https://doi.org/10.3390/diagnostics15131661 - 29 Jun 2025
Viewed by 493
Abstract
Background: Exophiala dermatitidis is a dematiaceous, thermotolerant, yeast-like fungus increasingly recognized as an opportunistic pathogen in chronic airway diseases. While commonly associated with cystic fibrosis, its clinical significance in non-cystic fibrosis bronchiectasis (NCFB) remains unclear. Case Presentation: We report the case of [...] Read more.
Background: Exophiala dermatitidis is a dematiaceous, thermotolerant, yeast-like fungus increasingly recognized as an opportunistic pathogen in chronic airway diseases. While commonly associated with cystic fibrosis, its clinical significance in non-cystic fibrosis bronchiectasis (NCFB) remains unclear. Case Presentation: We report the case of a 66-year-old immunocompetent woman with a history of breast cancer in remission and NCFB, who presented with chronic cough and dyspnea. Chest CT revealed bilateral bronchiectasis with new pseudonodular opacities. Bronchoalveolar lavage cultures identified E. dermatitidis, along with Pseudomonas aeruginosa and methicillin-sensitive Staphylococcus aureus. Given clinical stability and the absence of systemic signs, initial therapy included oral voriconazole, levofloxacin, doxycycline, and inhaled amikacin. Despite persistent fungal isolation on repeat bronchoscopy, the patient remained asymptomatic with stable radiologic and functional findings. Antifungal therapy was discontinued, and the patient continued under close monitoring. The patient exhibited clinical and radiological stability despite repeated fungal isolation, reinforcing the hypothesis of persistent colonization rather than active infection. Discussion: This case underscores the diagnostic challenges in distinguishing fungal colonization from true infection in structurally abnormal lungs. In NCFB, disrupted mucociliary clearance and microbial dysbiosis may facilitate fungal persistence, even in the absence of overt immunosuppression. The detection of E. dermatitidis should prompt a comprehensive evaluation, integrating clinical, radiologic, and microbiologic data to guide management. Voriconazole is currently the antifungal agent of choice, though therapeutic thresholds and duration remain undefined. Conclusions: This report highlights the potential role of E. dermatitidis as an under-recognized respiratory pathogen in NCFB and the importance of a multidisciplinary, individualized approach to diagnosis and treatment. This case underscores the need for further research on fungal colonization in NCFB and the development of evidence-based treatment guidelines. Further studies are needed to clarify the pathogenic significance, optimal management, and long-term outcomes of E. dermatitidis in non-CF chronic lung diseases. Full article
(This article belongs to the Section Diagnostic Microbiology and Infectious Disease)
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10 pages, 576 KiB  
Brief Report
First Molecular Evidence of Equine Herpesvirus Type 1 (EHV-1) in Ocular Swabs of Clinically Affected Horses
by Beatriz Musoles-Cuenca, Miguel Padilla-Blanco, Valentina Vitale, Teresa Lorenzo-Bermejo, María de la Cuesta-Torrado, Beatriz Ballester, Elisa Maiques, Consuelo Rubio-Guerri and Ana Velloso Alvarez
Viruses 2025, 17(6), 862; https://doi.org/10.3390/v17060862 - 18 Jun 2025
Viewed by 508
Abstract
Equine Herpesvirus Type 1 (EHV-1) is a significant pathogen within the Alphaherpesvirinae subfamily, causing respiratory disease, abortions, and, in severe cases, equine herpesvirus myeloencephalopathy (EHM). While nasal swabs and blood samples are commonly used for real-time polymerase chain reaction (RT-PCR) diagnosis, variability in [...] Read more.
Equine Herpesvirus Type 1 (EHV-1) is a significant pathogen within the Alphaherpesvirinae subfamily, causing respiratory disease, abortions, and, in severe cases, equine herpesvirus myeloencephalopathy (EHM). While nasal swabs and blood samples are commonly used for real-time polymerase chain reaction (RT-PCR) diagnosis, variability in viral shedding necessitates exploring additional sample types. This study reports the first molecular detection of EHV-1 in ocular swabs from naturally infected horses during an outbreak in the Valencian Community in 2023. Nasal and ocular swabs were collected from ten symptomatic horses and analyzed via RT-PCR. EHV-1 was detected in all cases, with higher viral loads in nasal samples. Although nasal swabs remain the most reliable sample for EHV-1 detection, the presence of viral DNA in tear fluid suggests a previously unrecognized route of viral shedding. These findings support further investigation into the role of ocular secretions in the pathogenesis and epidemiology of EHV-1. Additional studies are needed to determine the clinical relevance and potential utility of ocular swabs in specific outbreak scenarios. Full article
(This article belongs to the Special Issue Advances in Endemic and Emerging Viral Diseases in Livestock)
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22 pages, 782 KiB  
Review
From Infection to Autoimmunity: S. pyogenes as a Model Pathogen
by Virginia Girlando, Luisa De Angelis, Gianluca D’Egidio, Armando Di Ludovico and Luciana Breda
Microorganisms 2025, 13(6), 1398; https://doi.org/10.3390/microorganisms13061398 - 16 Jun 2025
Viewed by 814
Abstract
Group A β-hemolytic Streptococcus (GAS) is a Gram-positive, coccoid-shaped bacterium that tends to grow in chains; it is a non-spore-forming, facultatively anaerobic, catalase-negative, aerobic bacterium. It is known to cause a wide range of infections in children, ranging from mild upper respiratory tract [...] Read more.
Group A β-hemolytic Streptococcus (GAS) is a Gram-positive, coccoid-shaped bacterium that tends to grow in chains; it is a non-spore-forming, facultatively anaerobic, catalase-negative, aerobic bacterium. It is known to cause a wide range of infections in children, ranging from mild upper respiratory tract infections, such as pharyngitis, to severe invasive disease. GAS also notably triggers post-infectious immune sequelae, including acute poststreptococcal glomerulonephritis (APSGN), acute rheumatic fever (ARF), and rheumatic heart disease (RHD), which are major health burdens, especially in low-income countries. In this review, we will present the general characteristics of GAS, highlighting its structural and microbiological features. We will also discuss its pathogenetic mechanisms, especially molecular mimicry, and its ability to cause autoimmune responses. Finally, we will elucidate some of the autoimmune sequelae that mark GAS infections, such as ARF, RHD, APSGN, and guttate psoriasis. Understanding GAS as a model pathogen for infection-induced autoimmunity provides insight into host–pathogen interactions and supports the development of targeted interventions. Emphasis on early diagnosis and antibiotic treatment is essential to reduce the burden of autoimmune complications Full article
(This article belongs to the Special Issue Infections, Immune Mechanisms and Host-Pathogen Interactions)
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13 pages, 490 KiB  
Article
Clinical Significance of the 5T;12TG Genotype in Pediatric CFSPID: A Retrospective Study
by Ana Morales-Tirado, Enrique Blitz-Castro, Saioa Vicente-Santamaría, Carmen Luna-Paredes, Enrique Salcedo-Lobato, Ana Tabares-González, Celia Gascón-Galindo, Simon Boutry and Adelaida Lamas-Ferreiro
Children 2025, 12(6), 778; https://doi.org/10.3390/children12060778 - 14 Jun 2025
Viewed by 556
Abstract
Background: One of the most common genetic variants among individuals with cystic fibrosis screen-positive inconclusive diagnosis (CFSPID) is 5T;12TG. Classified as having “varying clinical consequences” (VVCC), it may produce a wide spectrum of CF phenotypes when combined in trans with a pathogenic variant [...] Read more.
Background: One of the most common genetic variants among individuals with cystic fibrosis screen-positive inconclusive diagnosis (CFSPID) is 5T;12TG. Classified as having “varying clinical consequences” (VVCC), it may produce a wide spectrum of CF phenotypes when combined in trans with a pathogenic variant on the other CFTR allele, ranging from asymptomatic cases to CFTR-related disorders (CFTR-RD) or classical cystic fibrosis (CF). The 5T;12TG variant is currently eligible for modulator treatment in the United States. Methods: We conducted a retrospective analysis of CFSPID children born between July 2009 and June 2023 in the Community of Madrid (Spain) who carried at least one 5T;12TG variant in trans with another CFTR variant. Data collected included trends in sweat chloride (SC) values, respiratory and digestive symptoms, lung function by spirometry, microbiological findings in nasopharyngeal aspirates, anthropometric data, and fecal elastase levels. Results: Twenty-one children (52.3% females; median age: 4.66 years [IQR 3.6–6.9]) were included. Eighteen had 5T;12TG in trans with a CF-causing variant (CFc), two had another VVCC variant, and one had a variant of unknown significance (VUS). After a median follow-up of 3.45 years [IQR 1.4–4.3], all the children remained asymptomatic. However, SC values rose to intermediate levels in nine (42.8%) of the children. No isolates of Pseudomonas aeruginosa were identified. Lung function and pancreatic markers remained normal. Conclusions: This is the first Spanish cohort of children with CFSPID and the 5T;12TG allele. Although clinical symptoms did not manifest during childhood, the SC value increased to intermediate values in 42.8% of the cohort, so these may require long-term follow-up to observe conversions to CFTR-RD or CF. The potential initiation of modulator therapy based solely on SC levels or emerging symptoms warrants careful consideration. Full article
(This article belongs to the Section Pediatric Pulmonary and Sleep Medicine)
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18 pages, 797 KiB  
Article
Risk of Incidence and Lethality by Etiology of Severe Acute Respiratory Syndrome in Hospitalized Children Under 1 Year of Age in Brazil in 2024: A Cross-Sectional Study
by Tamires de Nazaré Soares, Natasha Cristina Oliveira Andrade, Suziane do Socorro dos Santos, Marcela Raíssa Asevedo Dergan, Karina Faine Freitas Takeda, Jully Greyce Freitas de Paula Ramalho, Luany Rafaele da Conceição Cruz, Perla Katheleen Valente Corrêa, Marli de Oliveira Almeida, Joyce dos Santos Freitas, Wilker Alves Silva, Marcos Jessé Abrahão Silva, Daniele Melo Sardinha and Luana Nepomuceno Gondim Costa Lima
Trop. Med. Infect. Dis. 2025, 10(6), 168; https://doi.org/10.3390/tropicalmed10060168 - 14 Jun 2025
Viewed by 671
Abstract
Severe Acute Respiratory Syndrome (SARS) represents a significant cause of morbidity and mortality in children under one year of age, a particularly vulnerable population due to immunological and respiratory immaturity. The diverse etiology includes multiple respiratory viruses such as Respiratory Syncytial Virus (RSV), [...] Read more.
Severe Acute Respiratory Syndrome (SARS) represents a significant cause of morbidity and mortality in children under one year of age, a particularly vulnerable population due to immunological and respiratory immaturity. The diverse etiology includes multiple respiratory viruses such as Respiratory Syncytial Virus (RSV), influenza, rhinovirus, and SARS-CoV-2, each with distinct potential to cause severe illness and death. Understanding the specific incidence and lethality by etiological agents in the recent Brazilian context (2024), after the COVID-19 pandemic, is essential to guide surveillance and public health strategies. This study aimed to analyze the risk of incidence and lethality by specific etiology of SARS in children under one year of age hospitalized in Brazil during the year 2024. A descriptive cross-sectional study was performed using secondary data from the 2024 Influenza Epidemiological Surveillance Information System (SIVEP-Gripe), obtained via OpenDataSUS. Reported cases of SARS hospitalized in children <1 year of age in Brazil were included. Distribution by final classification and epidemiological week (EW) was analyzed; the incidence rate by Federative Unit (FU) (cases/100,000 < 1 year) with risk classification (Low/Moderate/High) was assessed; and, for cases with positive viral RT-PCR, the etiological frequency and virus-specific lethality rate (deaths/total cases of etiology ×100), also with risk classification, were extracted. A multivariate logistic regression model was performed for the risk factors of death. A total of 66,170 cases of SARS were reported in children under 1 year old (national incidence: 2663/100,000), with a seasonal peak between April and May. The majority of cases were classified as “SARS due to another respiratory virus” (49.06%) or “unspecified” (37.46%). Among 36,009 cases with positive RT-PCR, RSV (50.06%) and rhinovirus (26.97%) were the most frequent. The overall lethality in RT-PCR-positive cases was 1.28%. Viruses such as parainfluenza 4 (8.57%), influenza B (2.86%), parainfluenza 3 (2.49%), and SARS-CoV-2 (2.47%) had higher lethality. The multivariate model identified parainfluenza 4 (OR = 6.806), chronic kidney disease (OR = 3.820), immunodeficiency (OR = 3.680), Down Syndrome (OR = 3.590), heart disease (OR = 3.129), neurological disease (OR = 2.250), low O2 saturation (OR = 1.758), SARS-CoV-2 (OR = 1.569) and respiratory distress (OR = 1.390) as risk factors for death. Cough (OR = 0.477) and RSV (OR = 0.736) were associated with a lower chance of death. The model had good calibration (Hosmer–Lemeshow p = 0.693) and overall significance (p < 0.001). SARS represented a substantial burden of hospitalizations, with marked seasonal and geographic patterns. RSV and rhinovirus were the main agents responsible for the volume of confirmed cases but had a relatively low to moderate risk of lethality. In contrast, less frequent viruses such as parainfluenza 4, influenza B, parainfluenza 3, and SARS-CoV-2 were associated with a significantly higher risk of death. These findings highlight the importance of dissociating frequency from lethality and reinforce the need to strengthen etiological surveillance, improve diagnosis, and direct preventive strategies (such as immunizations) considering the specific risk of each pathogen for this vulnerable population. Full article
(This article belongs to the Special Issue Respiratory Infectious Disease Epidemiology and Control)
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10 pages, 741 KiB  
Article
Development of a qPCR Tool for Detection, Quantification, and Molecular Characterization of Infectious Laryngotracheitis Virus Variants in Chile from 2019 to 2023
by Tomás Gatica, Sebastián Salgado, Humberto Reyes and Carlos Loncoman
Animals 2025, 15(11), 1623; https://doi.org/10.3390/ani15111623 - 31 May 2025
Viewed by 492
Abstract
Infectious laryngotracheitis virus (ILT) is the pathogen responsible for a respiratory ailment that has resulted in significant economic losses in the poultry industry, primarily due to high morbidity and mortality rates, as well as diminished egg production. However, for small producers living near [...] Read more.
Infectious laryngotracheitis virus (ILT) is the pathogen responsible for a respiratory ailment that has resulted in significant economic losses in the poultry industry, primarily due to high morbidity and mortality rates, as well as diminished egg production. However, for small producers living near the coast, where backyard flocks are located in rural areas, respiratory illness may pose a public health risk rather than an economic one, which must be considered in the differential diagnosis of ILT. Therefore, in this research, we focused on developing an efficient tool to detect, quantify, and classify infectious laryngotracheitis virus (ILTV) field variants. Our results demonstrates that qPCR detected positive samples collected between 2019 and 2023 in flocks exhibiting clinical signs of respiratory illness in Valdivia, in the Los Ríos Region of Southern Chile. Furthermore, the molecular characterization of positive samples using dideoxynucleotide sequencing revealed that the detected viruses were similar to tissue culture origin (TCO) vaccines, even though the birds had never been vaccinated. To the best of our knowledge, this is the first report characterizing ILTV variants in Chile using a molecular approach. Our findings indicate that the tool is useful for detecting ILTV and can also be used to quantify viral particles using a standard curve, making it a valuable tool for the differential diagnosis of respiratory pathogens in poultry. Full article
(This article belongs to the Special Issue Common Infectious Diseases in Poultry)
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16 pages, 1109 KiB  
Article
Superinfections in Hospitalized COVID-19 Patients (Super COVID-19): Data from the Multicentric Retrospective CH-SUR Cohort Study in Switzerland
by Giulia Scanferla, Andrea Blöchlinger, Veronika Bättig, Michael Buettcher, Alexia Cusini, Anne Iten, Olivia Keiser, Rami Sommerstein, Jonathan Sobel and Werner C. Albrich
COVID 2025, 5(6), 86; https://doi.org/10.3390/covid5060086 - 30 May 2025
Viewed by 432
Abstract
Background: The epidemiology, characteristics and outcomes of coinfections in COVID-19 are still poorly understood. Methods: We investigated the prevalence of coinfections in COVID-19 patients hospitalized in Switzerland over the first three epidemic waves between 1 March 2020 and 1 June 2021, as well [...] Read more.
Background: The epidemiology, characteristics and outcomes of coinfections in COVID-19 are still poorly understood. Methods: We investigated the prevalence of coinfections in COVID-19 patients hospitalized in Switzerland over the first three epidemic waves between 1 March 2020 and 1 June 2021, as well as risk factors and outcomes. Patients were identified from six hospitals of the Swiss prospective surveillance system database (CH-SUR). Details of the type and treatment of coinfections were retrieved retrospectively from medical charts. We assessed the proportion of patients with suspected coinfections and analyzed risk factors and 90-day in-hospital survival using logistic and Cox regression. Results: Of 13,265 identified patients, 36.6% (4859/13,625) had suspected coinfections, and 44.8% (5941/13,625) received antibiotics. Respiratory coinfections (25.6%) were the most common, followed by bloodstream (19.8%) and urinary tract infections (14.6%). Escherichia coli (14.8%), Staphylococcus aureus (10.7%) and Klebsiella pneumoniae (6.1%) were the most frequently isolated pathogens. The risk factors for coinfections included increasing age, male gender, certain underlying medical conditions and immunosuppression. Suspected coinfections were associated with a longer hospital stay (13 vs. 7 days, p < 0.001), more frequent ICU admission (26% vs. 6.7%, p < 0.001) and higher rates of in-hospital death (24% vs. 9.5%, p < 0.001). Hospitalization in the ICU at the time of COVID-19 diagnosis had the strongest association with coinfections. Conclusions: A high proportion of COVID-19 patients had coinfections, particularly respiratory infections, and received antibiotics. Coinfections were associated with severe illness and worse outcomes. Full article
(This article belongs to the Section COVID Clinical Manifestations and Management)
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