E. bieneusi microsporidiosis is an opportunistic infection in immunocompromised patients. There is a lack of established treatment strategies. In this report, we present an overview of the available literature and three cases of E. bieneusi infection in renal transplant recipients.

To review the efficacy, safety, and clinical selection considerations of azole antifungal agents for prophylaxis of invasive fungal infections (IFIs) in immunocompromised patients using a clinically oriented, pharmacist-focused framework. A literature review was conducted using PubMed and Embase to identify relevant studies evaluating azole antifungal prophylaxis in immunocompromised adult populations. Search terms included combinations of "azole," "antifungal prophylaxis," "immunocompromised," and "invasive fungal infection," with emphasis on contemporary studies reflecting current prophylaxis practices. Studies were included if they evaluated prophylactic use of azole antifungal agents in immunocompromised adults, including patients with hematologic malignancies, hematopoietic stem cell transplantation, or other defined immunocompromised states. Studies focused on treatment rather than prophylaxis, non-azole agents, pediatric populations, or lacking relevant clinical outcomes were excluded. Data extracted included study design, patient population, antifungal agent and dosing, incidence of IFIs, and reported safety outcomes. Randomized controlled trials and observational studies demonstrate that azole antifungal prophylaxis significantly reduces IFI incidence in high-risk immunocompromised populations. Posaconazole has demonstrated superiority over fluconazole and itraconazole in neutropenic leukemia populations, while mold-active azoles including voriconazole and isavuconazole offer expanded prophylactic options in select patients. Therapeutic drug monitoring, drug interaction management, and individualized risk stratification are critical pharmacist-driven components of care. Safety findings remain consistent with known azole-associated toxicities including hepatotoxicity, QT interval alterations, neuropsychiatric effects, and cytochrome P450-mediated drug interactions. Azole antifungal prophylaxis remains a cornerstone strategy for prevention of IFIs in immunocompromised patients. Current evidence supports individualized, risk-based selection of prophylactic agents based on patient-specific risk factors rather than universal application of a single regimen. Pharmacists play a central role in optimizing prophylaxis through therapeutic drug monitoring, medication reconciliation, and toxicity mitigation. Additional studies are needed to optimize prophylaxis strategies and improve patient outcomes.

Preliminary study investigating the performance of the BACTECTM Mycosis IC/F blood culture system for Malassezia spp. detection.

Drug combination treatment as a strategy for inhibiting human adenovirus replication.

Scedosporium apiospermum is aubiquitous species of ascomycete fungi. Human infections occur mostly as opportunistic infections in immunocompromised patients. Risk factors for immunocompetent patients are penetrating trauma, surgery, and near-drowning accidents. We report acomplicated postsurgical infection with S.apiospermum in a66-year-old immunocompetent adult. After microsurgical lumbar decompression, the patient developed an extensive infection in the area of the operation with sub- and epidural abscess collection, spondylodiscitis, polyradiculitis, and arachnoiditis. S.apiospermum was recovered from multiple cultures of tissue samples as well as from implant material. Despite early initiation of voriconazole therapy and surgical intervention, the progressive infection of multiple lumbar nerve roots could not be prevented and the patient developed distal sensorimotor paraparesis. Only after 10months of continuous oral voriconazole therapy and repeated surgical procedures to replace all implant material did stabilization and clinical improvement occur. Finally, fungus was no longer detected and the patient slowly but steadily regained walking ability.

Hepatitis E virus (HEV) infection is generally self-limiting in immunocompetent individuals but may progress to chronic infection in immunocompromised patients, underscoring the need for effective antiviral therapies. Although ribavirin is currently used off-label for HEV treatment, its associated adverse effects highlight the need for safer alternatives. In this study, we screened an anti-viral compound library comprising 800 compounds using three HEV reporter systems designed to target distinct stages of the viral life cycle. Candidate compounds were further evaluated in PLC/PRF/5 cells using both acute and chronic infection models with wild-type genotype 3 HEV (HEV-3). Antiviral activity was assessed by measuring HEV RNA levels in culture supernatants. Elbasvir, a known inhibitor of hepatitis C virus (HCV) non-structural protein 5A (NS5A), was identified as the most potent candidate. Although multiple compounds showed inhibitory effects in reporter assays, only elbasvir achieved sustained suppression of HEV growth in long-term culture, reducing HEV RNA levels to below the limit of detection. In a chronic infection co-culture model, elbasvir maintained antiviral activity at non-cytotoxic concentrations. Time-of-addition analysis demonstrated that elbasvir inhibits an early step in the viral life cycle, specifically viral internalization. Furthermore, combination with ribavirin enhanced antiviral efficacy, resulting in sustained viral suppression without detectable cytotoxicity and exhibiting an additive interaction. Collectively, these findings identify elbasvir as a promising candidate for repurposing as an anti-HEV drug and support a combination strategy targeting distinct steps of the viral life cycle.

Investigation of a pseudo-outbreak of Burkholderia cepacia complex caused by contaminated phosphate-buffered saline.

Carbapenem-resistant Klebsiella pneumoniae (CRKP) is a multidrug-resistant (MDR) pathogen causing severe infections in immunocompromised patients, prompting the exploration of alternative therapies like bacteriophage therapy. In this study, we isolated and characterized a novel halophilic lytic bacteriophage, Halo KS-7, targeting K. pneumoniae, and used an AI-driven annotation pipeline in Python to analyze its genome and therapeutic potential. Bacteriophages were isolated from Hospital wastewater, purified through plaque isolation, and confirmed using the double-layer agar method. Morphological analysis via transmission electron microscopy (TEM) and plaque assays assessed lytic activity. In vitro assays, including one‑step growth curve and MOI determination, were performed to evaluate the replication kinetics and lytic activity of bacteriophage Halo KS‑7 against carbapenem‑resistant Klebsiella pneumoniae. In vivo efficacy was assessed using a BALB/c mouse wound infection model by monitoring wound contraction and performing blinded histopathological analysis following phage treatment. DNA sequencing was done using Illumina HiSeq 2000, followed by genome assembly, AI-guided annotation, gene prediction, protein function classification, and comparative genomics using CLC Genomics Workbench. We also evaluated host range, temperature stability, pH sensitivity, and salt stress tolerance to assess therapeutic potential. Halo KS-7 exhibited strong lytic activity against CRKP and was classified as a Myoviridae bacteriophage by TEM. Phenotypic assays demonstrated optimal activity at 37 °C and neutral pH, effective activity from pH 4–10, and enhanced performance in high-salinity conditions. Bacteriophage Halo KS-7 exhibited a short latent period (~20 min), a modest burst size (5.73 PFU/cell), and optimal antibacterial activity at MOI 0.1, resulting in sustained suppression of K. pneumoniae growth in vitro. In vivo, Halo KS-7 treatment significantly enhanced wound healing in infected BALB/c mice, achieving near-complete wound closure, effective infection control, and improved histopathological regeneration comparable to uninfected controls. Halo KS-7 have 58.716 kb linear dsDNA genome (44.4% G + C), contains 49 predicted ORFs, lacks integrase, lysogeny, or antibiotic-resistance genes, and includes three tRNA genes (tRNATyr, tRNAPro, and tRNAAsn). It also includes a toxin gene and auxiliary factors like MazG, pyrophosphatase, and HNH endonucleases that enhance bacterial killing without promoting horizontal gene transfer or resistance. Functional annotation assigned ~65% of ORFs to structural, replication, and packaging roles. Comparative genomics showed moderate similarity to other Myoviridae but with distinct accessory features, emphasizing its novelty and therapeutic value. Halo KS-7 is a novel, strictly lytic bacteriophage with strong antibacterial activity and stress resilience, supporting its use as a promising biocontrol agent against CRKP and its potential for clinical development in managing MDR infections.

Fungi are a group of microorganisms encompassing thousands of species, with only a minority causing disease in humans. Fungal pathogens and infections are increasing global public health problems. They are an important cause of opportunistic infections in immunocompromised patients; however, they can also affect immunocompetent individuals. This paper reviews the incidence, clinical features, radiographic manifestations, and pathological findings of major fungal infections in non-immunocompromised patients. Only a restricted subset of fungi, specifically the endemic dimorphic fungi, selected Aspergillus manifestations, and Cryptococcus gattii are consistently associated with pulmonary infection in immunocompetent individuals. Endemic mycoses are determined by their geographic distribution, which makes it necessary to consider the diagnosis when pulmonary infection occurs in endemic areas. However, their clinical and radiological manifestations are frequently nonspecific. On the other hand, other fungi, mainly Aspergillusspp. are involved in a variety of diseases, including allergic bronchopulmonary aspergillosis, which is frequently diagnosed based on imaging criteria, and a series of conditions in the spectrum of chronic aspergillosis, including simple aspergilloma, chronic cavitary pulmonary aspergillosis, chronic fibrosing pulmonary aspergillosis, subacute invasive pulmonary aspergillosis, and aspergillus nodules.

The pathogenic fungus Trichosporon asahii causes severe invasive fungal infections in immunocompromised patients with neutropenia. In Cryptococcus neoformans, calcineurin-responsive zinc finger 1 (Crz1) functions as a transcription factor downstream of the calcineurin signaling pathway and regulates the expression of genes involved in stress resistance and virulence. In T. asahii, Cna1 and Cnb1, which are key components of the calcineurin pathway, contribute to various stress responses and virulence. The role of Crz1 in stress tolerance and virulence in T. asahii, however, has remained unclear. Here, we demonstrate that a crz1 gene-deficient T. asahii mutant exhibited increased sensitivity to cell wall and endoplasmic reticulum stress. The crz1 gene-deficient mutant was sensitive to Congo red and tunicamycin but not to dithiothreitol or sodium dodecyl sulfate. Moreover, the virulence of the crz1 gene-deficient mutant in the silkworm infection model was reduced. These phenotypes of the crz1 gene-deficient mutant were restored by reintroducing the crz1 gene, confirming the association between Crz1 and these phenotypes. The half-maximal lethal dose of the cnb1 gene-deficient T. asahii mutant was higher than that of the crz1 gene-deficient mutant. These results suggest that Crz1 mediates the stress responses and virulence of T. asahii. The involvement of Crz1 in the virulence of T. asahii is small, however, compared with that of the calcineurin.

Abstract Opportunistic infections (OIs) remain a major cause of morbidity and mortality among immunocompromised individuals, and delayed or inaccurate diagnosis can rapidly lead to adverse clinical outcomes. This narrative review critically evaluates the diagnostic performance, clinical utility, and evolving applications of quantitative real-time polymerase chain reaction (qPCR) for detecting opportunistic pathogens in vulnerable patient populations. Evidence across bacterial, fungal, viral, and parasitic infections demonstrates that qPCR consistently surpasses conventional culture- and serology-based methods in sensitivity, specificity, and turnaround time, particularly in cases of low pathogen burden or prior antimicrobial exposure. Its quantitative capability enables dynamic pathogen load monitoring and facilitates timely, targeted therapeutic interventions. However, limitations such as assay variability, incomplete standardization, and challenges in distinguishing colonization from active infection in polymicrobial contexts persist. Ongoing advances in multiplex technologies, digital PCR, and integrative data-driven diagnostics are expected to further refine clinical implementation. Collectively, qPCR has evolved from a supplemental assay to a cornerstone technology in precision diagnostics for OIs in immunocompromised patients.

Cryptococcus neoformans is an opportunistic fungal pathogen that causes pulmonary infection in immunocompromised patients, which in severe cases leads to fatal meningoencephalitis. Cryptococcus exhibits unique glycobiology that plays important roles in pathogenesis. Unlike model yeast and other common fungal pathogens, Cryptococcus incorporates xylose, a five-carbon monosaccharide, into its glycans. One trimer motif, which consists of xylose in β-1,2 linkage to the reducing mannose of an α-1,3-mannose dimer, occurs in key cryptococcal glycoconjugates that include protein N- and O-linked glycans, glycosylinositol phosphorylceramides (GIPCs), and the capsule polysaccharides glucuronoxylomannan (GXM) and glucuronoxylomannogalactan (GXMGal). We previously identified cryptococcal β-1,2-xylosyltransferase 1 (Cxt1), which catalyzes formation of this motif in GIPCs, GXM, and GXMGal. Here, we report the discovery of a second enzyme, cryptococcal β-1,2-xylosyltransferase 2 (Cxt2). Through characterization of cells that lack one or both corresponding genes (CXT1 and CXT2), we have dissected the biological roles of these enzymes, which are overlapping but not identical. Notably, Cxt1 and Cxt2 co-localize in the Golgi, influence capsule in a strain-dependent manner, and together are responsible for all xylose addition to O-glycans. Overall, our work highlights unique roles of these two enzymes and fills a gap in understanding of cryptococcal glycan synthesis.

Pulmonary infections represent a significant global health concern, contributing substantially to morbidity and mortality worldwide. Metagenomic next-generation sequencing (mNGS) represents an advanced, comprehensive, and unbiased diagnostic approach for pathogen identification, effectively overcoming many limitations inherent in conventional diagnostic methods. This study aimed to systematically evaluate the clinical performance of mNGS in the etiological diagnosis of pulmonary infections, with a particular emphasis on its utility across diverse immune statuses. This retrospective study included 136 patients with suspected pulmonary infections admitted to the Department of Respiratory Medicine at Shandong Provincial Hospital from June 2023 to April 2025. Bronchoalveolar lavage fluid (BALF) samples were collected from all patients and concurrently subjected to mNGS and conventional microbiological testing (CMT). The pathogen detection spectrum and diagnostic performance of mNGS were systematically compared against those of CMT. mNGS exhibited a significantly higher overall pathogen detection rate compared to CMT (77.2% vs. 50.0%, P < 0.001). Regarding the pathogen spectrum, mNGS identified a broader array of microorganisms, encompassing 19 bacterial, 9 fungal, and 2 mycobacterial species, in contrast to the 11 bacterial, 5 fungal, and 1 mycobacterial species detected by CMT. Diagnostic performance analysis further revealed that mNGS sensitivity was significantly superior to that of CMT (74.6% vs. 46.7%, P < 0.001). Furthermore, mNGS demonstrated a distinct advantage in detecting mixed infections, with a detection rate of 19.1%, significantly exceeding that of CMT (8.8%, P < 0.05). Subgroup analysis indicated a significantly higher incidence of mixed infections in immunocompromised patients compared to immunocompetent patients (35.1% vs. 13.1%, P < 0.05). Additionally, immunocompromised patients were more frequently subjected to adjustments in antimicrobial therapy guided by mNGS results (56.8% vs. 35.4%, χ² = 5.094, P < 0.05). In conclusion, mNGS offers superior sensitivity and broader pathogen coverage for the etiological diagnosis of pulmonary infections compared to conventional microbiological testing. Its enhanced capability to detect mixed infections significantly improves diagnostic accuracy in immunocompromised patients and effectively facilitates the dynamic optimization of antimicrobial therapy. Serving as a powerful complement to traditional diagnostic methods, mNGS holds particular value for the rapid diagnosis of complex and immunosuppression-associated pulmonary infections.

Introduction Hepatitis E virus (HEV) is a widespread, while epidemiologically underassessed, single-stranded, positive-sense RNA virus. Of the four main genotypes, genotypes 1 and 2 occur rather in developing countries, genotype 3 is most common in Europe, and genotype 4 causes infections in East Asia. Risk factors for infection between genotypes include sanitary conditions and drinking contaminated water for genotypes 1 and 2, and consumption of undercooked pork or wild boar meat for genotypes 3 and 4. Receiving blood transfusions is also associated with the risk of transmission. Hepatitis E virus has recently become the subject of interest beyond the field of infectious disease specialists, as the focus is gradually shifting towards its occurrence in the immunocompromised. It is important to raise awareness among healthcare professionals who may encounter patients with this condition. Therefore, this review aims to provide an up-to-date synthesis of the literature on the topic. Material and Methods The Scopus database was queried with the following search terms: “hepatitis E virus”, “HEV”, “rheumatology”, “connective tissue disease”. Results Of the 277 results obtained, 105 articles were then selected to be included in the review. Discussion While HEV infection may be asymptomatic, the typical course involves acute hepatitis. Chronic infections also occur, especially in immunocompromised individuals, which also involves those treated for rheumatologic diseases. Genotype 3 and 4 infections are associated with extrahepatic manifestations, such as neurological symptoms, renal complications, acute pancreatitis, cryoglobulinemia, and vasculitis. Molecular mimicry is the proposed mechanism underlying these conditions. The infection is usually self-limiting and does not require treatment. However, patients undergoing immunosuppressive therapy may need a dose reduction or even complete withdrawal from the therapy. Off-label use of ribavirin may be considered. Conclusions The HEV infection in immunocompromised patients may be severe, leading to diagnostic challenges, severe complications, and treatment dilemmas. To establish optimal prevention and treatment strategies, further research is required

Pneumocystis pneumonia (PcP) caused by Pneumocystis jirovecii is one of the most common and serious opportunistic infections in immunocompromised patients. People at risk for developing Pneumocystis pneumonia include human immunodeficiency virus positive individuals, cancer patients, people receiving immunosuppressive therapy, organ transplant recipients and people with compromised immune systems. Despite effective treatment and prophylaxis, mortality is reported to be between 15-40%, even in people with immunodeficiency other than acquired immunodeficiency syndrome. Although the number of immunosuppressed patients is increasing, diagnostic and epidemiological studies for this agent are still insufficient. This study aimed to investigate the prevalance and epidemiological characteristics of P.jirovecii isolated from various patient groups. Among the 469 bronchoalveolar lavage fluids or sputum samples sent to the medical microbiology laboratory from various units, 114 samples were selected for indirect fluorescent antibody (IFA) testing and molecular studies based on an examination of the patients' files to determine underlying diseases, use of immunosuppressive drugs, corticosteroid treatments received and the presence of patchy or nodular ground-glass opacity on computed tomography scans and the presence of a cyst-like structure on May-Grunwald Giemsa staining. The presence of P.jirovecii was investigated by IFA and polymerase chain reaction methods and genotyping was performed by sequencing of mtLSU rRNA and internal transcribed spacer (ITS) region. The obtained ITS gene region DNA sequences were aligned and TCS network analysis was performed in the aligned region. As a result, positivity was found in eight samples out of 469 samples by using mtLSU rRNA genotyping, Genotype 1 (n= 3; 37.5%) was found to be the most common genotype in our samples. Genotype 2 (n= 2, 25%), genotype 4 (n= 2, 25%) and genotype 3 were the most frequently detected other genotypes, respectively. When the relationships between the sequences were examined, it was observed that our samples were generally related to the samples originating from Iran. In the present study, the genotyping analysis of the ITS region, constructed using the consensus sequence employed in the study by Lee et al., revealed that the most common genotype was Eh (n=3; 42.85%), followed by Bg (n= 2; 28.57%), Bi (n= 1; 14.28%) and Eg (n=1; 14.28%). Our study is the first genotyping study conducted in our country using the ITS gene region. Different epidemiological findings were found in P.jirovecii genotype frequencies in studies conducted in different geographies. This suggests that genetic variations in P.jirovecii have a geographical component and this may affect the distribution of P.jirovecii strains among humans. According to TCS network analysis, our samples are generally associated with samples originating from Iran. None of our samples are found alongside samples originating from India. However, in certain areas of the analysis, New World and Old-World samples exist together. It can be assumed that globalization and thus the increase in human movement over time has led to the spread of different genotypes to different geographical regions and the formation of genotypic mosaics in certain geographical regions. Man-made destruction of nature and the consequent intertwining of urban and rural boundaries, as well as global warming and climate change have undeniably contributed to these movements.

Acinetobacter baumannii is a leading cause of healthcare-associated infections in immunocompromised patients and frequently exhibits multidrug resistance. Cefiderocol, a siderophore cephalosporin, is among the few remaining therapeutic options for infections caused by carbapenem-resistant A. baumannii (CRAB); however, its activity may differ by clonal lineage and can be further compromised in the biofilm state. This study investigates genomic features and cefiderocol efficacy against planktonic and biofilm-associated forms of oncology-derived A. baumannii isolates. Twenty-five non-duplicate, consecutive clinical isolates of A. baumannii from oncology patients underwent whole-genome sequencing and multilocus sequence typing. Cefiderocol activity was quantified in planktonic and biofilm-associated states using minimum bactericidal concentration (MBC) and minimum biofilm eradication concentration (MBEC) assays. Ten sequence types were identified, with the high-risk sequence type 2 (ST2) clone accounting for 56% (14/25) of isolates. ST2 strains showed significantly higher resistance to aminoglycosides, carbapenems, and fluoroquinolones than non-ST2 (NST) strains. The carbapenemase gene bla OXA-23 was detected exclusively in ST2. Colistin and cefiderocol were the most active agents overall. ST2 strains showed higher cefiderocol MBC values than NST strains. However, avibactam significantly reduced cefiderocol MBC in ST2, consistent with class D β-lactamases activity. ST2 and NST isolates exhibited comparable distributions of iron acquisition genes and similar CAS-detected siderophore activity under the assay conditions tested. Cefiderocol activity was significantly reduced in biofilms relative to planktonic cells (median MBEC 2 µg/ml versus median MBC 0.5 µg/ml). NST exhibited higher MBEC/MBC ratios than ST2 isolates, indicating greater biofilm-associated tolerance to cefiderocol. Collectively, these data associate the predominance of oncology-derived ST2 with bla OXA-23 carriage and higher cefiderocol bactericidal thresholds and show that cefiderocol activity is consistently reduced in the biofilm state. Future studies integrating functional measures of iron acquisition and β-lactamase activity will be needed to define the determinants of cefiderocol efficacy across lineages and growth states.

Background Timely aetiological diagnosis of pulmonary infection in immunocompromised patients (ICPs) remains challenging because clinical presentations may be atypical and conventional microbiological tests (CMTs) have limited sensitivity. Probe capture based targeted next generation sequencing (ptNGS) has emerged as a potential alternative to metagenomic next generation sequencing (mNGS), but its clinical performance in this population remains incompletely defined. Methods In this cross-sectional diagnostic accuracy study, immunocompromised adults undergoing bronchoalveolar lavage for suspected pulmonary infection were enrolled. Bronchoalveolar lavage fluid (BALF) samples were analysed using CMTs, mNGS, and ptNGS. Composite clinical adjudication served as the reference standard. Diagnostic performance was compared at the case level, and pulmonary microbiota characteristics were explored. Results Among 78 enrolled patients, 60 were classified as having pulmonary infection. Causative pathogens were identified in 52 cases, and fungal pathogens, particularly Pneumocystis jirovecii, were the most frequently detected. At the case level, ptNGS and mNGS demonstrated higher sensitivity than CMTs (80.0% vs 80.0% vs 26.7%) and showed high concordance in microorganisms identified (91.7%). Specificity was 72.2% for CMTs, compared with 44.4% for mNGS and 38.9% for ptNGS. Positive sequencing results were also observed in patients without pulmonary infection (n = 18), predominantly involving viral or opportunistic microorganisms. Microbiota analysis of 65 samples revealed reduced microbial alpha diversity and altered community composition in patients with pulmonary infection. Conclusions In ICPs with suspected pulmonary infection, ptNGS substantially increases pathogen detection compared with CMTs and demonstrates diagnostic performance comparable to mNGS. Sequencing results require careful clinical interpretation, given the difficulty in distinguishing infection from colonisation in respiratory specimens. Exploratory microbiota analyses suggest infection associated alterations in lung microbial ecology that warrant further validation.

Cytomegalovirus (CMV) causes the most prevalent and severe opportunistic infection in immunocompromised patients following solid organ or hematopoietic stem cell transplantation, with the highest morbidity and mortality rates among herpesviruses. The study objective was to compare and determine the CMV chronic infection and related hematological and immunological markers in immunodeficient and immunocompetent participants. A prospective case–control study among 85 participants was designed to measure CMV-immunoglobulin G (IgG) and to evaluate interleukin-34 and serum calprotectin as biomarkers; total leukocyte, granulocyte, lymphocyte, and platelet counts were also measured following blood collection. A high CMV IgG positivity was observed across all groups in this investigation, indicating widespread chronic infection. CMV IgG, interleukin-34 (IL-34), and calprotectin levels did not differ significantly between immunocompetent and immunocompromised individuals. There was no significant association between CMV IgG and IL-34 or serum calprotectin. Furthermore, IL-34 showed a significantly higher mean in males compared to females (p = 0.002). An exploratory observation was that IL-34 had a moderate positive correlation with serum calprotectin (ρ = 0.609, 95% confidence interval: 0.450–0.731, p &lt; 0.001) across all study participants. The study findings call for more research to elucidate the clinical roles of calprotectin and IL-34 in immunocompromised patients.

Background:Diagnosing invasive mold infections (IMI) is challenging because they typically occur as opportunistic infections in immunocompromised patients who often present with non-specific symptoms. Furthermore, no single test can definitively diagnose IMI, and a proven diagnosis often requires invasive sampling. This sampling can be unsafe and difficult to perform, especially in the immunocompromised population most at risk for these infections.Objectives:The objective of this study is to assess the clinical utility of plasma microbial cell-free DNA next-generation sequencing (mcfDNA-NGS) for diagnosing invasive mold infections in the context of conventional diagnostic methods.Design:Retrospective observational study at a quaternary care center (2017–2025).Methods:The charts of 30 patients with mold-positive mcfDNA-NGS (Karius™ Spectrum; Redwood City, CA, USA) were reviewed, with IMI adjudicated per 2020 EORTC/MSGERC criteria. Provider documentation, medication orders, and patient outcomes were used to assess clinician interpretation of mcfDNA-NGS result accuracy (true positive vs false positive) and its impact on diagnostic and therapeutic decision-making. Turnaround time (TAT) and molecules per microliter (MPM) were summarized.Results:IMI final classifications were proven (6), probable (8), possible (5), and unclassified (11). Overall, 23/30 (77%) results were true positives. Among these 23 patients with true positive results for mold, 5 (22%) received a new clinical adjudication of IMI that had been entirely missed by conventional diagnostic testing. Furthermore, mcfDNA NGS provided species-level pathogen identification in 9 of the 23 (39%) true positive cases where conventional testing detected fungal elements or elevated biomarkers but could not identify the specific organism. Median TAT was 102 h; median MPM 657. MPM did not differ between true and false positives (p = 0.86). mcfDNA-NGS changed diagnostic classification in 7/30 (23%) and antimicrobial management in 16/30 (53%).Conclusion:mcfDNA-NGS provided noninvasive, actionable information, informing diagnosis and therapy. Future studies should define optimal stewardship and cost-effectiveness.

Aspergillus fumigatus leads to serious systemic infections in immunocompromised patients, thereby emphasizing the need for novel immunomodulatory therapies acting on both innate (sCD14) and adaptive (IL-21) immune systems. Beta-glucans are established fungal immunomodulators, and diphtheria toxoid (DT) has non-specific stimulatory activity. This experiment determined the effects of beta-glucan, biphasic beta-glucan, and DT on the systemic immune response to A. fumigatus beta-glucan, DT, and a combination of both. It was done on 50 male albino rats, and they were then subjected to a systemic injection of A. fumigatus challenge. Serum IL-21 and soluble CD14 (sCD14) levels were measured on day 21 using the enzyme-linked immunosorbent assay kits. The results revealed a significant increase in IL-21 and sCD14 levels in the beta-glucan group compared with those in the control group. Nevertheless, the beta-glucan + DT group had the most potent effect, with an increase in both markers. The combination of beta-glucan and diphtheria toxoid provides a potent immunomodulatory regimen that boosts innate (sCD14) and adaptive (IL-21) immune pathways and enhances host defense against systemic fungal infection.