Search Results (33)

Background: Polymerase chain reaction (PCR) is highly sensitive and specific for the rapid diagnosis of invasive fungal disease (IFD) but is not yet widely implemented due to concerns regarding limited standardisation between assays, the lack of commercial options and the absence of clear guidance on interpreting results. Objectives and Methods: This review provides an update on technical and clinical aspects of PCR for the diagnosis of the most pertinent fungal pathogens, including Aspergillus, Candida, Pneumocystis jirovecii, Mucorales spp., and endemic mycoses. Summary: Recent meta-analyses have demonstrated that quantitative PCR (qPCR) offers high sensitivity for diagnosing IFD, surpassing conventional microscopy, culture and most serological tests. The reported specificity of qPCR is likely underestimated due to comparison with imperfect reference standards with variable sensitivity. Although the very low limit of detection of qPCR can generate false positive results due to procedural contamination or patient colonisation (particularly in pulmonary specimens), the rates are comparable to those observed for biomarker testing. When interpreting qPCR results, it is essential to consider the pre-test probability, determined by the patient population, host factors, clinical presentation and risk factors. For patients with low to moderate pre-test probability, the use of sensitive molecular tests, often in conjunction with serological testing or biomarkers, can effectively exclude IFD when all tests return negative results, reducing the need for empirical antifungal therapy. Conversely, for patients with high pre-test probability and clinical features of IFD, qPCR testing on invasive specimens from the site of infection (such as tissue or bronchoalveolar lavage fluid) can confidently rule in the disease. The development of next-generation sequencing methods to detect fungal infection has the potential to enhance the diagnosis of IFD, but standardisation and optimisation are essential, with improved accessibility underpinning clinical utility. Full article

The wild boar (Sus scrofa) has experienced significant population growth as well as geographic expansion across Europe over the past 15 years, leading to increased concerns regarding its role in the transmission of zoonotic pathogens. Among these, Babesia spp. and Anaplasma spp. are of particular importance due to their impact on both wildlife and domestic animals. This study systematically reviews the prevalence and distribution of Babesia and Anaplasma spp. in wild boars and associated tick vectors across multiple European countries, synthesizing data from literature published between 2010 and 2024. A comprehensive search of Scopus, Google Scholar, and PubMed databases was conducted using predefined keywords related to babesiosis, anaplasmosis, wild boars, Europe, and tick-borne diseases. A total of 281 studies were initially retrieved, of which 19 met the inclusion criteria following relevance assessment. Data extraction focused on pathogen identification, diagnostic methods, sample type, host species, and prevalence rates. Molecular detection methods, primarily PCR and sequencing, were the most used diagnostic tools. Results indicate substantial regional variations in the prevalence of Babesia and Anaplasma spp. A. phagocytophilum was detected in wild boar populations across multiple countries, with the highest prevalence rates observed in Slovakia (28.2%) and Poland (20.34%). Conversely, lower prevalence rates were recorded in France (2%) and Portugal (3.1%). Babesia spp. showed higher prevalence rates in Italy (6.2%), while its detection in other regions such as Romania and Spain was minimal or absent. Notably, spleen and multi-organ samples (spleen/liver/kidney) exhibited higher positivity rates compared to blood samples, suggesting an organotropic localization of these pathogens. The findings underscore the role of wild boars as reservoirs for tick-borne pathogens and highlight their potential to contribute to the epidemiological cycle of these infections. The increasing distribution of wild boars, coupled with climate-driven shifts in tick populations, may further facilitate pathogen transmission. Future studies should focus on integrating molecular, serological, and ecological approaches to improve surveillance and risk assessment. Standardized methodologies across different regions will be essential in enhancing comparative epidemiological insights and informing targeted disease management strategies. Full article

This study aims to investigate the prevalence of E. coli and E. coli O157:H7 in 353 samples collected in Kirkuk from human stool, animal feces, raw and pasteurized milk, and beef hamburgers. E. coli was isolated using conventional methods and identified with the Enterosystem Kit 18R. Suspected E. coli O157:H7 were confirmed serologically and tested for antimicrobial resistance and virulence genes (stx1, stx2, eaeA, and hlyA). The overall prevalence rates of 20.4% for E. coli and 7.9% for E. coli O157:H7 were found, with the highest prevalence in human stool. The antimicrobial susceptibility profile of 28 E. coli O157:H7 isolates revealed significant resistance and sensitivity patterns, highlighting important implications for public health. The isolates demonstrated complete sensitivity to gentamicin (100%), while also showing high sensitivity to ciprofloxacin (92.86%), ceftriaxone (85.71%), and amikacin (64.29%). Conversely, the isolates exhibited notable resistance to tetracycline (85.71%), ampicillin (75.00%), sulfamethoxazole (71.43%), and streptomycin (67.86%). All the E. coli O157:H7 strains isolated in this study were positive for stx1 and/or stx2, as well as the eaeA gene, and are referred to as enterohemorrhagic (EHEC) strains. In order to highlight the genotypic variability among the EHEC E. coli O157:H7 isolates, five virulence profiles were identified, with profile III (stx2, eaeA, and hlyA) being the most common (35.7%). This profile was closely associated with diarrheic humans, while profile V (stx1, eaeA) was prevalent in animal feces and products. These findings may raise awareness of the risks associated with this pathogen, helping to reduce the incidence of E. coli-related diseases and to protect human health. Full article

Alimentary tract inflammation in inflammatory bowel disease (IBD) is treated by systemically administered drugs that alter fundamental host immune responses. Biologics that target tumor necrosis factor (TNF) are first-line biologics in IBD, used widely for their effectiveness, steroid-sparing quality, and lower cost. While they enable a significant proportion of patients to achieve clinical remission, they carry an increased risk of infection and poor serological responses to vaccination. Conversely, our understanding of adaptive T cell responses in anti-TNF-treated IBD patients remains limited. The introduction of COVID-19 vaccines has prompted research that both challenges and refines our view on immunomodulatory therapy and its potential implications for immunity and protection. Here, we review these emergent findings, evaluate how they shape our understanding of vaccine-induced T cell responses in the context of anti-TNF therapy in IBD, and provide a perspective highlighting the need for a holistic evaluation of both cellular and humoral immunity in this population. Full article

Between 2013 and 2016, the A/H1N1pdm09 component of the live attenuated influenza vaccine (LAIV) produced instances of lower-than-expected vaccine effectiveness. Standard pre-clinical ferret models, using a human-like vaccine dose and focusing on antigenic match to circulating wildtype (wt) strains, were unable to predict these fluctuations. By optimising the vaccine dose and utilising clinically relevant endpoints, we aimed to develop a ferret efficacy model able to reproduce clinical observations. Ferrets were intranasally vaccinated with 4 Log₁₀ FFU/animal (1000-fold reduction compared to clinical dose) of seven historical LAIV formulations with known (19–90%) H1N1 vaccine efficacy or effectiveness (VE). Following homologous H1N1 wt virus challenge, protection was assessed based on primary endpoints of wt virus shedding in the upper respiratory tract and the development of fever. LAIV formulations with high (82–90%) H1N1 VE provided significant protection from wt challenge, while formulations with reduced (19–32%) VE tended not to provide significant protection. The strongest correlation observed was between reduction in wt shedding and VE (R² = 0.75). Conversely, serum immunogenicity following vaccination was not a reliable indicator of protection (R² = 0.37). This demonstrated that, by optimisation of the vaccine dose and the use of non-serological, clinically relevant protection endpoints, the ferret model could successfully translate clinical H1N1 LAIV VE data. Full article

Helicobacter pylori (H. pylori) has been implicated in colorectal carcinogenesis. Here, the association of immune responses to bacterial exposure with advancing stages of colorectal neoplasia was assessed by multiplex serology. Immunoglobulin (Ig) A and G antibody responses to thirteen proteins of H. pylori were measured by a Luminex-based multiplex assay in plasma from patients with colorectal cancer (CRC, n = 25), advanced adenoma (n = 82), or small polyps (n = 85) and controls (n = 100). Multivariable logistic regression was used to assess the association of bacterial seropositivity with colorectal neoplasia. The threshold for overall seropositivity required subjects to be positive for at least 4 out of the 13 tested antigens. In a cohort subset with matched data (n = 34), H. pylori seropositivity was correlated with bacterial abundance in both neoplastic and matched normal tissue. While no association was found between H. pylori seropositivity and the presence of CRC, IgA seropositivity to CagA was associated with a decreased risk of advanced adenoma (odds ratio, OR = 0.48, 95% confidence intervals, CIs: 0.24–0.96). Regarding IgG, higher antibody responses to HpaA was associated with advanced adenoma occurrence (OR = 2.46, 95% CI: 1.00–6.01), while responses to HP0395, CagA and Catalase were associated with polyp development (OR = 2.65, 95%, CI: 1.31–5.36, OR = 1.83, 95% CI: 1.01–3.32, and OR = 2.16, CI: 1.09–4.29, respectively). Positive correlations were found between H. pylori abundance in the normal mucosa and levels of both the IgA and IgG antibody response to Catalase and VacA antigens (r = 0.48, p < 0.01; r = 0.37, p = 0.04; r = 0.51, p < 0.01; and r = 0.71, p = 0.04, respectively). Conversely, H. pylori abundance was negatively correlated with levels of IgA antibody response to HpaA and with IgG antibody response to HP0231 in the diseased tissue (r = −0.34, p = 0.04 and r = −0.41, p = 0.01, respectively). The association between levels of H. pylori antigens and colorectal neoplasia risk gradually decreased with the adenoma progression, implicating the early activation of the immune response at the polyp stage. Thus, the evaluation of antibody response to certain bacterial antigens may indicate the presence of early-stage colorectal neoplasia. Further studies are needed to clarify the role H. pylori or the immune response to its antigens may have in colorectal carcinogenesis stages. Full article

Leptospirosis is a widespread zoonosis recognised as a re-emerging infectious disease in both humans and dogs, yet the actual seroprevalence of Leptospira in pets in Italy is relatively unknown. The aim of this study was to evaluate Leptospira antibody prevalence in dogs and cats from a shelter by the microscopic agglutination test (MAT), the gold standard test in leptospiral serology, and to assess risk factors for Leptospira infection. This seroepidemiological study investigated the prevalence of leptospiral antibodies in a cohort of 106 dogs and 51 cats housed in a municipal shelter in Milan. Blood samples were collected from the animals during two sampling periods: spring/summer 2014 and autumn/winter 2016/2017. Eight serogroups were evaluated: L. Australis, L. Ballum, L. Canicola, L. Grippotyphosa, L. Icterohaemorrhagiae, L. Pomona, L. Sejroe, and L. Tarassovi. Antibody titres ranged from 1:100 to 1:6400. The results indicated that 21.7% of dogs had antibodies against serogroups L. Icterohaemorrhagiae and L. Australis, making them the most often found. Conversely, none of the cats showed any presence of antibodies. Seropositivity was higher in the spring/summer period (32.7%) than in autumn/winter (11.1%), and no statistically significant results were found regarding sex or age. These findings underscore the importance of ongoing serological surveillance and biosecurity measures in shelter environments to mitigate the zoonotic risk posed by leptospirosis. Full article

Babesiosis, caused by Babesia ovis, is a major seasonal issue in sheep, particularly in countries like Türkiye with high Rhipicephalus bursa tick populations. Previous studies employing various methods such as microscopy, serology, or molecular techniques have reported different epidemiological data concerning ovine babesiosis. Addressing this knowledge gap, our study employed a combined nested PCR (nPCR)/indirect ELISA (iELISA) approach, analyzing blood samples collected from 414 sheep between April and July 2023 using both techniques. nPCR amplified the 18S ribosomal RNA gene of B. ovis and determined a molecular prevalence of 1.9%. Conversely, serological testing using iELISA targeted the BoSA1 antigen and revealed a significantly higher positivity rate of 59.9% for anti-B. ovis antibodies. The temporary presence of Babesia after recovery reduces nPCR sensitivity, resulting in lower molecular prevalence. However, even if Babesia is not present in the host, anti-B. ovis antibodies remain in the serum for a long time and can be detected serologically. Our study underscores the necessity of concurrently employing molecular and serological methods for an accurate assessment of B. ovis prevalence. It highlights the importance of comprehensive epidemiological approaches for effective disease management in sheep populations. Full article

The African swine fever virus (ASFV) mutant ASFV-G-∆I177L is a safe and efficacious vaccine which induces protection against the challenge of its parental virus, the Georgia 2010 isolate. Although a genetic DIVA (differentiation between infected and vaccinated animals) assay has been developed for this vaccine, still there is not a serological DIVA test for differentiating between animals vaccinated with ASFV-G-∆I177L and those infected with wild-type viruses. In this report, we describe the development of the ASFV-G-∆I177L mutant having deleted the EP402R gene, which encodes for the viral protein responsible for mediating the hemadsorption of swine erythrocytes. The resulting virus, ASFV-G-∆I177L/∆EP402R, does not have a decreased ability to replicates in swine macrophages when compared with the parental ASFV-G-∆I177L. Domestic pigs intramuscularly (IM) inoculated with either 10² or 10⁶ HAD₅₀ of ASFV-G-∆I177L/∆EP402R remained clinically normal, when compared with a group of mock-vaccinated animals, indicating the absence of residual virulence. Interestingly, an infectious virus could not be detected in the blood samples of the ASFV-G-∆I177L/∆EP402R-inoculated animals in either group at any of the time points tested. Furthermore, while all of the mock-inoculated animals presented a quick and lethal clinical form of ASF after the intramuscular inoculation challenge with 10² HAD₅₀ of highly virulent parental field isolate Georgia 2010 (ASFV-G), all of the ASFV-G-∆I177L/∆EP402R-inoculated animals were protected, remaining clinically normal until the end of the observational period. Most of the ASFV-G-∆I177L/∆EP402R-inoculated pigs developed strong virus-specific antibody responses against viral antigens, reaching maximum levels at 28 days post inoculation. Importantly, all of the sera collected at that time point in the ASFV-G-∆I177L/∆EP402R-inoculated pigs did not react in a direct ELISA coated with the recombinant EP402R protein. Conversely, the EP402R protein was readily recognized by the pool of sera from the animals immunized with recombinant live attenuated vaccine candidates ASFV-G-∆I177L, ASFV-G-∆MGF, or ASFV-G-∆9GL/∆UK. Therefore, ASFV-G-∆I177L/∆EP402R is a novel, safe and efficacious candidate with potential to be used as an antigenically DIVA vaccine. Full article

Breast cancer is a prevalent malignancy in the present day, particularly affecting women as one of the most common forms of cancer. A significant portion of patients initially present with localized disease, for which curative treatments are pursued. Conversely, another substantial segment is diagnosed with metastatic disease, which has a worse prognosis. Recent years have witnessed a profound transformation in the prognosis for this latter group, primarily due to the discovery of various biomarkers and the emergence of targeted therapies. These biomarkers, encompassing serological, histological, and genetic indicators, have demonstrated their value across multiple aspects of breast cancer management. They play crucial roles in initial diagnosis, aiding in the detection of relapses during follow-up, guiding the application of targeted treatments, and offering valuable insights for prognostic stratification, especially for highly aggressive tumor types. Molecular markers have now become the keystone of metastatic breast cancer diagnosis, given the diverse array of chemotherapy options and treatment modalities available. These markers signify a transformative shift in the arsenal of therapeutic options against breast cancer. Their diagnostic precision enables the categorization of tumors with elevated risks of recurrence, increased aggressiveness, and heightened mortality. Furthermore, the existence of therapies tailored to target specific molecular anomalies triggers a cascade of changes in tumor behavior. Therefore, the primary objective of this article is to offer a comprehensive review of the clinical, diagnostic, prognostic, and therapeutic utility of the principal biomarkers currently in use, as well as of their clinical impact on metastatic breast cancer. In doing so, our goal is to contribute to a more profound comprehension of this complex disease and, ultimately, to enhance patient outcomes through more precise and effective treatment strategies. Full article

In response to the COVID-19 pandemic, German public health authorities launched various infection control procedures. In line with this, anti-pandemic infection control was also implemented for German military and police deployments. The presented study assessed the impact of this increased infection control effort on deployment-associated infections in a holistic approach. To do so, the results of post-deployment assessments offered to German soldiers and police officers at the Department of Tropical Medicine and Infectious Diseases of the Bundeswehr Hospital Hamburg obtained during the pandemic period were compared to the results recorded during the pre-pandemic period in an exploratory, hypothesis-forming comparative study. In total, data from 1010 military deployments and 134 police deployments, predominantly to the African or the Eastern Mediterranean WHO regions, were included in the analyses. In the main results, a significant decrease in gastroenteritis in deployed soldiers (20.1% versus 61.3%, p < 0.0001) and at least a trend in the same direction in deployed police officers (25.7% versus 35.4%, p = 0.4026) were shown for the pandemic period, while no consistent tendency into the one or the other direction was detectable for febrile illness on deployment. In contrast to the finding of less frequently reported deployment-associated gastroenteritis, the detection rates of enteric microorganisms after deployment, including poor hygiene-related colonization with apathogenic protozoa, remained unchanged. Regarding non-enteric infections, the numbers of serologically confirmed malaria cases on deployment and as expected, due to increased airway protection, Mycobacterium tuberculosis-specific immune-conversion dropped significantly with p = 0.0037 and p = 0.009, respectively. As a side finding, soldiers and police officers with post-deployment medical assessments were more likely to be older and male during the pandemic compared to the pre-pandemic period. In summary, only minor changes in deployment-associated infection and colonization rates were seen in response to the increased infection control procedures during the pandemic period, apart from respiratory infections. In particular, the clinical finding of less gastroenteritis on deployment was not matched by a concordant decline in poor hygiene-related enteric colonization with apathogenic protozoa in the soldiers’ guts, indicating that the fecal–oral transmission risk remained basically the same. Full article

Background: Rheumatoid Arthritis (RA) is a systemic and chronic autoimmune disease with inflammation at the synovial joints. The purposes of this study were to evaluate the correlation between serological variables and temporomandibular disorders (TMDs) in patients diagnosed with RA, evaluated through diagnostic criteria for temporomandibular disorders (DC/TMD), and to check the influence of comorbidities on the different TMD signs and symptoms, or any serological variables. Methods: This observational cohort research study included seventeen patients affected by RA. The comorbidities and some variables from the initial serological analyses were collected from the clinical rheumatological charts. Then, the presence of any of the following TMD signs/symptoms, temporal myalgia, temporomandibular joint arthralgia, click or crepitus, was evaluated through the symptom questionnaire of the DC/TMD during clinical evaluation following the DC/TMD examination form. Results: Rheumatoid factor (RF), anticitrullinated protein antibodies (ACPA), and anti-nuclear antibody (ANA) positivity were present in 82.4%, 52.9%, and 41.2% of patients, respectively. Indicators of tissue inflammation were evaluated with median values of 21 mm/h for erythrocyte sedimentation rate (ESR) and 0.50 mg/dL for C-reactive protein (CRP). The RA patients presented comorbidities such as hypertension in 70.6% and diabetes in 23.5%. Negative ACPA had a significant association with myalgia (p = 0.03), and positive ANA was significantly associated with crepitus (p = 0.05). Conclusion: ANA and ACPA evaluation can be considered predictive serological tests associated with specific TMDs. Conversely, no influence of any comorbidities was found between different TMD signs and symptoms, or any serological variables. Full article

Foot-and-mouth disease (FMD) is a fatal contagious viral disease that affects cloven-hoofed animals and causes severe economic damage at the national level. There are seven serotypes of the causative foot-and-mouth disease virus (FMDV), and type O is responsible for serious outbreaks and shows a high incidence. Recently, the Cathay, Southeast Asia (SEA), and ME-SA (Middle East-South Asia) topotypes of type O have been found to frequently occur in Asia. Thus, it is necessary to develop candidate vaccines that afford protection against these three different topotypes. In this study, an experimental FMD vaccine was produced using a recombinant virus (TWN-JC) with the JC epitope (VP1 140–160 sequence of the O/SKR/Jincheon/2014) between amino acid 152 and 153 of VP1 in TWN-R. Immunization with this novel vaccine candidate was found to effectively protect mice against challenge with the three different topotype viruses. Neutralizing antibody titers were considerably higher after a second vaccination. The serological differences between the topotype strains were identified in guinea pigs and swine. In conclusion, a significant serological difference was observed at 56 days post-vaccination between animals that received the TWN-JC vaccine candidate and those that received the positive control virus (TWN-R). The TWN-JC vaccine candidate induced IFNγ and IL-12B. Full article

Human immunoglobulin allotypes are allelic antigenic determinants (or “markers”) determined serologically, classically by hemagglutination inhibition, on the human immunoglobulin (IG) or antibody heavy and light chains. The allotypes have been identified on the gamma1, gamma2, gamma3, and alpha2 heavy chains (designated as G1m, G2m, G3m, and A2m allotypes, respectively) and on the kappa light chain (Km allotypes). Gm and Am allotypes have been one of the most powerful tools in population genetics, as they are inherited in fixed combinations, or Gm–Am haplotypes, owing to the linkage of the human IGHC genes in the IGH locus on chromosome 14. They have been very instrumental in molecular characterization of the human IGHC genes (gene polymorphisms or alleles, and IG heavy-chain structure in domains) and of the IGH locus (IGHC gene order, gene conversion, and copy number variation (CNV)). They represent a major system for understanding immunogenicity of the polymorphic IG chains in relation to amino acid and conformational changes. The WHO/IMGT allotype nomenclature and the IMGT unique numbering for constant (C) domain bridge Gm–Am and Km alleles to IGHC and IGKC gene alleles and structures and, by definition, to IG chain immunogenicity, opening the way for immunoinformatics of personalized therapeutic antibodies and engineered variants. Full article

Understanding antibody persistence concerning multimorbidity is crucial for vaccination policies. Our goal is to assess the link between multimorbidity and serological response to SARS-CoV-2 nine months post-first vaccine. We analyzed Healthcare Workers (HCWs) from three cohorts from Italy, and one each from Germany, Romania, Slovakia, and Spain. Seven groups of chronic diseases were analyzed. We included 2941 HCWs (78.5% female, 73.4% ≥ 40 years old). Multimorbidity was present in 6.9% of HCWs. The prevalence of each chronic condition ranged between 1.9% (cancer) to 10.3% (allergies). Two regression models were fitted, one considering the chronic conditions groups and the other considering whether HCWs had diseases from ≥2 groups. Multimorbidity was present in 6.9% of HCWs, and higher 9-months post-vaccine anti-S levels were significantly associated with having received three doses of the vaccine (RR = 2.45, CI = 1.92–3.13) and with having a prior COVID-19 infection (RR = 2.30, CI = 2.15–2.46). Conversely, lower levels were associated with higher age (RR = 0.94, CI = 0.91–0.96), more time since the last vaccine dose (RR = 0.95, CI = 0.94–0.96), and multimorbidity (RR = 0.89, CI = 0.80–1.00). Hypertension is significantly associated with lower anti-S levels (RR = 0.87, CI = 0.80–0.95). The serological response to vaccines is more inadequate in individuals with multimorbidity. Full article