Diagnosis Of Invasive Fungal Infections Research Articles

A-294 Exocytosis Inhibitor Increases yield of Limulus Amebocyte Lysate (LAL), Used for adjunct diagnosis of Invasive Fungal infection

Abstract Background The limulus amebocyte lysate (LAL) was widely used in endotoxin detection in vitro and the clinical diagnosis of invasive fungal infection. The previous data showed that a caffeine collection solution could prevent degranulation during the blood collection procedure, and caffeine-derived LAL could work in both the chromogenic test and turbidimetric method assays. However, the cell pellets collected by caffeine collection solution aggregates after re-suspension in 5 mM CaCl2. No similar phenomenon was observed with PBS collection buffer. The purpose of this study is to set up a PBS-caffeine bleed solution to improve the yield of LAL Methods During the 2023 bleed season, experiments were carried out to test the results of PBS-caffeine solution on the degranulation during bleed process. 6 crabs were bled once a week for a total of 60 crabs by the PBS-caffeine collection solution. The blood of each crab was collected by PBS caffeine collection solution. The cell pellets were finally re-suspended into 5 mM CaCl2 to promote degranulation. The LAL activity was tested by both chromogenic and turbidimetric assay methods. Results The data demonstrates that PBS-caffeine collection solution could prevent degranulation for over 4 hours, and the activity yield of PBS-caffeine-derived LAL was higher. There was no discernable impact on the activity yield of the crab’s size during the bleed season. PBS-caffeine-collection solution could reduce amebocyte aggregation/clot formation during processing. Studies described the biochemical characteristics of PBS-caffeine-derived LAL. Moreover, PBS-caffeine-derived LAL could work in both the chromogenic and turbidimetric assays. Conclusions First, PBS-caffeine collection solutions prevent amebocyte degranulation during blood collection and processing. Next, a PBS-caffeine collection solution could reduce cell aggregation/clotting during processing, and the activity of PBS-caffeine LAL is higher. Finally, PBS LAL is functional in both the chromogenic test and turbidimetric assays

A-232 Characterize and reduce positive interference of hemolysis on Fungitell testing for (1, 3)-β-glucan (BDG) in serum

Abstract Background Fungitell is an FDA cleared in vitro diagnostic product with an indication for use as an adjunct test for the diagnosis of invasive fungal infection. This assay has been established using a kinetic photometric method. A key aspect of this (1→3)-β-glucan (BDG) activated cascade reaction is its characteristic kinetic optical density profile. Interference which affects the BDG kinetic profile makes data analysis and interpretation difficult. This interference is roughly classified as being associated with hemolysis, icterus, lipemia (collectively HIL), and other non-specific factors. Methods In this study, we found that the use of 5% PEG-8000 supplemented serum (1:1 ratio of the volume of serum to 10% PEG-8000 solution with 0.1 M Tris-Cl, pH 7.4), heated at 80°C for 3.5 min, followed by centrifugation at 15,000 x g for 10 minutes (PEG-X Method) can largely eliminate the hemolysis-associated interference. Results The treated serum Fungitell BDG reaction curves are more like the ideal BDG kinetic profile. The recovery of introduced β-glucan has been shown to be acceptable (80-120%) for hemolyzed specimens. Conclusions This simple and rapid procedure shows promise as a pre-analytical treatment to reduce serum interference in the Fungitell β-glucan test for a substantial fraction of samples that produce currently unreportable results.

A-274 The Yield of the Limulus Amebocyte Lysate (LAL), Which is Used for Endotoxin Detection and Fungal Diagnosis is Increased by Caffeine Collection Buffer

Abstract Background Limulus Amebocyte Lysate (LAL), which is a major component of Fungal infection diagnosis kit, is used for the adjunct diagnosis of invasive fungal infection. This study used different blood collection buffers to find the optimal blood collection buffer to maximize the yield of LAL. The PBS collection buffer was found to prevent degranulation during blood collection procedure and the PBS-derived Limulus Amebocyte Lysate (LAL) was observed to work in both the chromogenic test and the turbidimetric method assays. A similar phenomenon was observed with the caffeine collection buffer. Methods To further confirm this observation, we used a caffeine collection buffer to collect the blood of horseshoe crabs in the 2023 bleeding season (from July to October). Once a week, 3 crabs were bled for a total of 30 crabs. The blood of each crab was collected using the caffeine collection buffer and the 3% NaCl collection buffer, respectively. The cell pellets were resuspended into LAL reagent water (LRW) or 5 mM CaCl2. The final LAL activity was tested by both chromogenic and turbidimetric assay methods. Results The results showed that a caffeine collection buffer could prevent degranulation for over 1 hr and the activity yield of caffeine-derived LAL is much higher than that of the 3% NaCl solution. There was no discernible impact upon activity yield of crab size, period of the bleed season, or the vendors. Interestingly, caffeine-derived LAL was observed to work in both the chromogenic and turbidimetric assays. The enzyme characteristic of the LAL was also described. Conclusions Caffeine collection buffer prevents amebocyte degranulation during blood collection and processing. The activity of caffeine LAL is much higher than that of 3% NaCl. Caffeine LAL works in both chromogenic test and turbidimetric assay.

Revision of antifungal strategies definitions for invasive fungal infections (proven/probable/possible) in 461 patients with haematological malignancies (REDEFI-SEIFEM).

Invasive fungal infections (IFI) are a relevant cause of morbidity and mortality among patients with haematological neoplasms (HMs). Since 2002, a classification of IFI based on host factors, clinical and radiological features and mycological tests was published for research purpose. These criteria are widely used in clinical practice to identify patients at risk for IFI. The aim of the study was to evaluate the clinical applicability of EORTC/MSG 2008 criteria for the diagnosis of IFI in daily practice. This multicentre, non-interventional, observational, prospective study gathered all consecutive inpatients with HMs in which an intravenous antifungal treatment was started. Exclusion criteria were a previous or concomitant transplant procedure, outpatient status and oral antifungal therapy. EORTC/MSG 2008 criteria were used to classify patients at the beginning of antifungal therapy and at 30 days. An independent board reviewed the classification of IFI given by local clinicians at T0 and T30. The highest percentage of agreement was found for possible IFI (96%), while a lower agreement was reported for proven IFI (74%), and the highest variability was observed for probable IFI (56%). At T30, the board re-evaluation confirmed a strict agreement for possible IFI only (98%). Among 306 patients classified as possible, 156 (51%) patients showed non-typical radiological findings and 45 (15%) patients presented host factors only. In real life, the EORTC/MSG criteria can be applicable only for possible IFI. As non-typical radiological findings are reported in possible IFI, introducing a new IFI category should be considered.

A retrospective Italian analysis on the characteristics of invasive fungal infections in the intensive care unit setting: CHARTER-IFI study.

Invasive fungal infections (IFI), prevalent in critically ill ICU patients, have gained attention due to post-COVID-19 epidemiological shifts. Notably, COVID-19-associated aspergillosis and candidiasis pose significant risks. WHO recognises key fungal pathogens, emphasising the need for enhanced research and interventions. The CHARTER-IFI study retrospectively examines 186,310 individuals admitted to ICUs in Italy from 01/01/2012-01/09/2023, utilising administrative databases covering around 10 million inhabitants. Adult patients were included having at least one ICU discharge diagnosis of IFI at their first IFI-related hospitalisation and having at least 12 months of available data prior to this hospitalisation. A total of 746 IFI patients discharged from ICU (incidence of 4.0 per 1000 ICU-hospitalised patients), were included. Median age was 68 years, 63% were males, and the overall Charlson Comorbidity Index was 2.2. The top three diagnoses were candidiasis (N = 501, 2.7/1000 ICU-hospitalised patients), aspergillosis (N = 71, 0.4/1000), and pneumocystosis (N = 55, 0.3/1000). The evaluation of the comorbidity profile in IFI patients revealed the presence of hypertension (60.5%), use of systemic GC/antibacterials (45.3% during 12 months before and 18.6% during 3 months before hospital admission), cancer (23.1%), diabetes (24.3%) and cardiovascular diseases (23.9%). The mean (±SD) length of hospitalisation in ICU was 19.9 ± 24.1 days (median 11 days), and deaths occurred in 36.1% of IFI patients (within 30 days from discharge). This retrospective analysis among ICU-hospitalised patients described the burden of IFI in ICU, and its understanding could be crucial to strengthen surveillance, investments in research, and public health interventions as required by WHO.

A Diagnostic Stewardship Intervention to Improve Utilization of 1,3 β-D-Glucan Testing at a Single Academic Center: Five-Year Experience.

(1,3)- β-D-glucan (BDG) testing is one of the noninvasive tests to aid diagnosis of invasive fungal infections (IFIs). The study results have been heterogenous, and diagnostic performance varies depending on the risks for IFI. Thus, it is important to select appropriate patients for BDG testing to prevent false-positive results. An algorithmic diagnostic stewardship intervention was instituted at a single academic medical center to improve BDG test utilization. The BDG test order in the electronic health record was replaced with the BDG test request order, which required approval to process the actual test order. The approval criteria were (1) immunocompromised or intensive care unit patient and (2) on empiric antifungal therapy, or inability to undergo invasive diagnostic procedures. A retrospective observational study was conducted to evaluate the efficacy of the intervention by comparing the number of BDG tests performed between 1 year pre- and post-intervention. Safety was assessed by chart review of the patients for whom BDG test requests were deemed inappropriate and rejected. The number of BDG tests performed per year decreased by 85% from 156 in the pre-intervention period to 24 in the post-intervention period. The average monthly number of BDG tests performed was significantly lower between those periods (P = .002). There was no delay in IFI diagnosis or IFI-related deaths in the patients whose BDG test requests were rejected. The sustained effectiveness of the intervention was observed for 5 years. Institution of the diagnostic stewardship intervention successfully and safely improved BDG test utilization.

Hope on the Horizon? Aptamers in Diagnosis of Invasive Fungal Infections.

Despite remarkable advances in the diagnosis of invasive fungal infections (IFIs), rapid, specific, sensitive, and cost-effective detection methods remain elusive. Due to their stability, ease of production, and specificity to signature molecules of fungal pathogens, short single-stranded sequences of DNA, RNA, and XNA, collectively called aptamers, have emerged as promising diagnostic markers. In this perspective, we summarize recent progress in aptamer-based diagnostic tools for IFIs and discuss how these tools could potentially meet the needs and provide economical and simple solutions for point-of-care for better management of IFIs.

A review of the epidemiology of invasive fungal infections in Asian patients with hematological malignancies (2011-2021).

The incidence of invasive fungal infection (IFI) is increasing, especially among patients diagnosed with hematological malignancies due to their immunocompromised nature. Other risk factors include advanced age, exposure to immunosuppressants, neutropenia, and catheter use. Some of the most common IFI organisms reported are Candida and Aspergillus species, and other fungal species, including Scedosporium, Trichosporon, Cryptococcus, and Fusarium have also increasingly been reported in the past years. However, the epidemiologic data on IFI among patients with hematological malignancies in Asian countries are lacking. Therefore, we investigated published epidemiologic data on such cases from the past 10 years (2011-2021) and discuss the challenges faced in the diagnosis and management of IFIs in Asia.

Exploring European Consensus About the Remaining Treatment Challenges and Subsequent Opportunities to Improve the Management of Invasive Fungal Infection (IFI) in the Intensive Care Unit

BackgroundThe global prevalence of invasive fungal infections (IFI) is increasing, particularly within Intensive Care Units (ICU), where Candida spp. and Aspergillus spp. represent the most important pathogens. Diagnosis and management of IFIs becomes progressively challenging, with increasing antifungal resistance and the emergence of rare fungal species.Through a consensus survey focused on assessing current views on how IFI should be managed, the aim of this project was to identify challenges around diagnosing and managing IFIs in the ICU. The current status in different countries and perceived challenges to date amongst a multidisciplinary cohort of healthcare professionals involved in the care of IFI in the ICU was assessed.MethodsUsing a modified Delphi approach, an expert panel developed 44 Likert-scale statements across 6 key domains concerning patient screening and minimal standards for diagnosis of IFIs in ICU; initiation and termination of antifungal treatments and how to minimise their side effects and insights for future research on this topic. These were used to develop an online survey which was distributed on a convenience sampling basis utilising the subscriber list held by an independent provider (M3 Global). This survey was distributed to intensivists, infectious disease specialists, microbiologists and antimicrobial/ICU pharmacists within the UK, Germany, Spain, France and Italy. The threshold for consensus was set at 75%.ResultsA total of 335 responses were received during the five-month collection period. From these, 29/44 (66%) statements attained very high agreement (≥ 90%), 11/44 (25%) high agreement (< 90% and ≥ 75%), and 4/44 (9%) did not meet threshold for consensus (< 75%). ConclusionThe results outline the need for physicians to be aware of the local incidence of IFI and the associated rate of azole resistance in their ICUs. Where high clinical suspicion exists, treatment should start immediately and prior to receiving the results from any diagnostic test. Beta-D-glucan testing should be available to all ICU centres, with results available within 48 h to inform the cessation of empirical antifungal therapy. These consensus statements and proposed measures may guide future areas for further research to optimise the management of IFIs in the ICU.

The effect of antifungal treatments on bronchoalveolar lavage galactomannan for the diagnosis of invasive fungal infections in patients with hematological malignancy: Single-center experience

Background Invasive fungal infections (IFI) mainly caused by aspergillus species are one of the leading causes of death in patients undergoing immunosuppressive therapy for hematological malignancies. The presence of galactomannan in bronchoalveolar lavage fluid (BAL GM) is an important diagnostic marker. Some of the factors affecting the BAL GM are still unknown. Antifungal treatment administered before or after BAL also affect BAL GM results. Methods To investigate the effect of BAL GM timing on the diagnosis of IFE, 100 patients receiving immunosuppressive therapy with hematological malignancy at the Uludağ University Faculty of Medicine, Hematology Department, and underwent BAL over a 3-year period with the suspicion of IFE as well as 127 BAL procedures of these patients were examined. Results There were 70 patients who started antifungal therapy before BAL and 30 patients who did not. BAL GM was found positive in 33 (47.1%) of the 70 patients who received antifungal therapy compared to 22 (73.3%) of the 30 patients who did not receive antifungal therapy. There was a significant difference between the two groups in terms of BAL GM positivity (p = 0.016). Subsequently, 127 BAL procedures of these 100 patients were evaluated. When the second, third, and subsequent BAL procedures of the same patients were included in the study, BAL GM was positive in 41 (46.6%) of the 88 procedures in patients who received treatment before BAL and in 25 (64.1%) of the 39 procedures in patients who did not receive treatment before BAL. The rate of BAL GM positivity did not differ between groups (p = 0.068). Conclusions The balance between reducing the risk by initiating early antifungal therapy and maximizing the diagnostic value of BAL GM should be evaluated individually for each patient.

Valuable Role of Beta-D-Glucan in The Diagnosis of Invasive Fungal Infections in Patients with Hematological Malignancies

Valuable Role of Beta-D-Glucan in The Diagnosis of Invasive Fungal Infections in Patients with Hematological Malignancies

Disseminated Histoplasmosis in a Patient with Myelofibrosis on Ruxolitinib: A Case Report and Review of the Literature on Ruxolitinib-Associated Invasive Fungal Infections.

Ruxolitinib, a selective inhibitor of Janus kinases, is a standard treatment for intermediate/high-risk myelofibrosis (MF) but is associated with a predisposition to opportunistic infections, especially herpes zoster. However, the incidence and characteristics of invasive fungal infections (IFIs) in these patients remain uncertain. In this report, we present the case of a 59-year-old woman with MF who developed disseminated histoplasmosis after seven months of ruxolitinib use. The patient clinically improved after ten weeks of combined amphotericin B and azole therapy, and ruxolitinib was discontinued. Later, the patient received fedratinib, a relatively JAK2-selective inhibitor, without relapse of histoplasmosis. We also reviewed the literature on published cases of proven IFIs in patients with MF who received ruxolitinib. Including ours, we identified 28 such cases, most commonly due to Cryptococcus species (46%). IFIs were most commonly disseminated (39%), followed by localized lung (21%) infections. Although uncommon, a high index of suspicion for opportunistic IFIs is needed in patients receiving JAK inhibitors. Furthermore, the paucity of data regarding the optimal management of IFIs in patients treated with JAK inhibitors underscore the need for well-designed studies to evaluate the epidemiology, pathobiology, early diagnosis, and multimodal therapy of IFIs in patients with hematological malignancies receiving targeted therapies.

INTERPRETABLE MACHINE LEARNING FOR PREDICTING RISK OF INVASIVE FUNGAL INFECTION IN CRITICALLY ILL PATIENTS IN THE INTENSIVE CARE UNIT: A RETROSPECTIVE COHORT STUDY BASED ON MIMIC-IV DATABASE.

The delayed diagnosis of invasive fungal infection (IFI) is highly correlated with poor prognosis in patients. Early identification of high-risk patients with invasive fungal infections and timely implementation of targeted measures is beneficial for patients. The objective of this study was to develop a machine learning-based predictive model for invasive fungal infection in patients during their intensive care unit (ICU) stay. Retrospective data was extracted from adult patients in the MIMIC-IV database who spent a minimum of 48 h in the ICU. Feature selection was performed using LASSO regression, and the dataset was balanced using the BL-SMOTE approach. Predictive models were built using six machine learning algorithms. The Shapley additive explanation algorithm was used to assess the impact of various clinical features in the optimal model, enhancing interpretability. The study included 26,346 ICU patients, of whom 379 (1.44%) were diagnosed with invasive fungal infection. The predictive model was developed using 20 risk factors, and the dataset was balanced using the borderline-SMOTE (BL-SMOTE) algorithm. The BL-SMOTE random forest model demonstrated the highest predictive performance (area under curve = 0.88, 95% CI = 0.84-0.91). Shapley additive explanation analysis revealed that the three most influential clinical features in the BL-SMOTE random forest model were dialysis treatment, APSIII scores, and liver disease. The machine learning model provides a reliable tool for predicting the occurrence of IFI in ICU patients. The BL-SMOTE random forest model, based on 20 risk factors, exhibited superior predictive performance and can assist clinicians in early assessment of IFI occurrence in ICU patients. Importance: Invasive fungal infections are characterized by high incidence and high mortality rates characteristics. In this study, we developed a clinical prediction model for invasive fungal infections in critically ill patients based on machine learning algorithms. The results show that the machine learning model based on 20 clinical features has good predictive value.

A Seminested PCR Method for the Diagnosis of Invasive Fungal Infections in Combat Injured.

Among combat injured, invasive fungal infections (IFIs) result in significant morbidity. Cultures and histopathology are the primary diagnostic methods for IFIs, but they have limitations. We previously evaluated a panfungal polymerase chain reaction assay, which was 83% sensitive and 99% specific for angioinvasive IFIs. Here, we evaluated 3 less resource-intensive seminested assays targeting clinically relevant fungi in the order Mucorales and genera Aspergillus and Fusarium. Formalin-fixed paraffin-embedded tissue specimens from a multicenter trauma IFI cohort (2009-2014) were used. Cases were US military personnel injured in Afghanistan with histopathologic IFI evidence. Controls were patients with similar injury patterns and no laboratory IFI evidence (negative culture and histopathology). Seminested assays specific to Mucorales (V4/V5 regions of 18S rDNA), Aspergillus (mitochondrial tRNA), and Fusarium (internal transcribed spacer [ITS]/28A regions of DNA) were compared with a panfungal assay amplifying the internal transcribed spacer 2 region of rDNA and to histopathology. Specimens from 92 injury sites (62 subjects) were compared with control specimens from 117 injuries (101 subjects). We observed substantial agreement between the seminested and panfungal assays overall, especially for the order Mucorales. Moderate agreement was observed at the genus level for Aspergillus and Fusarium. When compared with histopathology, sensitivity and specificity of seminested assays were 67.4% and 96.6%, respectively (sensitivity increased to 91.7% when restricted to sites with angioinvasion). Prior studies of seminested molecular diagnostics have focused on culture-negative samples from immunocompromised patients. Our findings underscore the utility of the seminested approach in diagnosing soft-tissue IFIs using formalin-fixed paraffin-embedded tissue samples, especially with angioinvasion.

Coadministration of isavuconazole and sirolimus in allogeneic hematopoietic stem cell transplant recipients.

Invasive fungal infections (IFIs) represent a major cause of morbidity among allogeneic hematopoietic stem cell transplantation (allo-HSCT). Isavuconazole (ISA) is a broad-spectrum triazole with favorable safety profile. Herein, we evaluate the real life coadministration of ISA and sirolimus in allo-HSCT recipients in a single-center retrospective analysis, describing clinical efficacy, safety, and therapeutic drug monitoring (TDM) of both drugs. All consecutive allo-HSCT recipients who received the coadministration of ISA and sirolimus for at least 2 weeks between July 2017 and December 2022 were included in this retrospective analysis. TDM was longitudinally performed during treatment. IFIs were classified according to the revised European Organization for Research and Treatment of Cancer/Mycoses Study Group consensus criteria. A total of 51 recipients were included in the analysis. A total of 17 patients received ISA as continuous antifungal treatment for IFI diagnosed before transplant: one patient experienced a probable invasive pulmonary aspergillosis, and one patient switched from ISA to liposomal amphotericin B for a possible IFI. A total of 34 patients started ISA as antifungal therapy for IFI diagnosed after transplant. Sixteen of 34 were treated for a proven/probable breakthrough IFI during mold-active prophylaxis: 6/16 patients died for IFI after a median of 51 days of ISA. Eighteen of 34 started ISA as empirical therapy for a possible IFI: 15/18 patients were alive with resolution of infection after 6 weeks, 1 died for disease progression, and 2 had empirically changed antifungal therapy due to pneumonia progression. Clinical and radiological response rate was 68% after 90 days from IFI diagnosis. No toxicities related to drug-drug interaction have been registered in patients reaching concomitant therapeutic levels of ISA and sirolimus. The coadministration of ISA and sirolimus was safe and feasible in this cohort, confirming favorable clinical efficacy in patients with multiple-drug coadministration.

Benefits of early antifungal therapy in hematology patients

Invasive fungal infections (IFIs) are a serious threat to patients with hematological diseases. These infections are characterized by high mortality and lead to significant financial costs for treatment. The most common pathogens of IFIs are Aspergillus spp. and Candida spp., but in recent years, cases of infections caused by rare pathogens have become more frequent. Diagnosis of IFIs and choice of treatment remain challenging due to the nonspecificity of symptoms and the diversity of clinical cases. In this regard, the problem of start time and choice of antifungal therapy remains of current interest. This review briefly describes diagnostic criteria, challenges associated with IFIs diagnosing, provides evidence for empiric and preventive strategies as two early treatment approaches, and examines the impact of therapy initiation on patient outcomes. Treatment of IFIs in hematologic patients should be individualized. At the same time, early administration of therapy with broad-spectrum drugs for febrile neutropenia and parallel diagnostic measures can improve treatment outcomes. There is a lack of current data on the benefits of specific treatment strategies, highlighting the need for further research.

Invasive fungal infection in patients with hematologic malignancies: epidemiology and prognostic factors.

Invasive fungal infections (IFI) are emerging opportunistic diseases that occur mainly in immunocompromised patients. Our study aimed to analyze the epidemiology of IFIs in patients with hematological malignancies, and the prognostic factors. Our retrospective study included patients hospitalized in the hematology department between January 1st, 2010, and August 31st, 2020, and in whom the diagnosis of IFI was made according to the EORTC criteria 2008. We found 29 IFIs among 6989 admissions (0.4%). IFIs were proven in 16 cases and probable in 13 cases. The median age was 35 years. The sex ratio was 0.9. The predominant IFI was invasive pulmonary aspergillosis (n=14) followed by fungemia (n=13). Candida albicans was the most isolated species in blood cultures (5/9). The mortality rate was 48%. In multivariate analysis, disease status, time to start antifungal treatment, and lactate levels are significant factors of excess mortality. IFIs are responsible for significant morbidity and mortality. The challenge lies in the precocity of starting the treatment as well as the vigilance given to the factors of poor prognosis.

Epidemiology and Clinical Outcomes of Hematologic Patients with Proven and Probable Invasive Fungal Infections: A Latin American Tertiary Care Centre Experience

Introduction Although diagnostic and therapeutic strategies for invasive fungal infections (IFI) have evolved in the last decade, their identification and management within patients with hematologic diseases (HD) continues to be a challenge. For this group, in addition to the increased risk of developing an IFI, mortality has been reported as high as 39.1%. The incidence and etiology vary across regions, suggesting an influence of patient-specific characteristics, environment, and clinician's practices. Literature regarding low- and middle-income countries, reflecting the real-world approach and management for this high-risk group is scarce. Our aim was to describe patients' clinical and microbiological characteristics, treatment strategies, and outcomes in a tertiary care hospital in Mexico City. Methods Retrospective single-center study that included all consecutive cases of IFI occurring in adult patients with HD or hematopoietic stem cell transplant (HSCT) between January 2013 and December 2019. We included only proven and probable cases of IFI according to the mycoses study group of the European Organization for Research and Treatment of Cancer (EORTC) guidelines. Clinical data, diagnostic work-up, treatment modalities, and outcomes were extracted from the medical records. Results In a total of 119 patients, 129 IFIs were diagnosed; criteria were fulfilled for proven in 38% ( n = 49) and probable in 62% ( n = 80). The median age was 39 years (range, 18-87) and HD was acute leukemia at 46.5%. At the time of IFI, 83.7% had active HD and 82.2% were on active treatment. HSCTs were allogeneic in 11 patients and autologous in 2 patients. A 31.8% and 91.5% presented with at least one comorbidity and/or risk factor, respectively. Severe neutropenia was present in 67.4% with a median of 13 days (range, 0-167) from the start of neutropenia to IFI diagnosis. Additional characteristics are shown in Table 1. The median time from the HD/HSCT to IFI was 73 days (range, 0-1,598); 30.2% being diagnosed within the treatment induction phase. Identified pathogenswere Aspergillus sp in 62.4%, Candida sp in 21.3%, Mucor spin10.6%, Fusarium sp in 3.5%, and Histoplasma capsulatum in 2.1%. In 10 (7.8%) cases, fungal coinfection was identified. For proven cases, sites where the microorganism was isolated or identifiedwere: blood 48.3%, lung 25.9%, sinuses 13.8%, CSF 3.4%, bone marrow 3.4%, skin 1.8%,and others 3.4%. For probable IFI, sitesof infection were: lung 91.8%, sinuses 5.9%, and other 2.4%. A 92.8% of aspergillosis and 80% of mucormycosis infections were localized in the lungs, while 86.7% of candidiasis were identified in the blood. Forty-five (35.4%) patients were receiving antifungal prophylaxis (fluconazole n = 24, itraconazole n = 13, posaconazole n = 3, voriconazole n = 3, anidulafungin n = 2). Antifungal treatment was given to24 (96.1%) of patients; 5 died before IFI diagnosis. Frontline agents included voriconazole ( n = 62), amphotericin n = 26), anidulafungin ( n = 16), caspofungin ( n = 6), fluconazole ( n = 5), clinical trial (posaconazole vs. voriconazole, n = 5) itraconazole ( n = 3), and posaconazole ( n = 1). Overall survival (OS) at 6 weeks was 59.7% and at 12 weeks was 51.9%. Overall response rate (ORR) was 48.4% ( n=60) and 54.8% ( n=68) at 6 and 12 weeks, respectively. On univariate analysis risk factors associated with increased mortality at 12-weeks were: age &amp;gt;40 ( p=0.001), active HD ( p = 0.020), CVC 30 days prior to IFI ( p=0.032), IMV 30 days prior to IFI ( p=0.026), and admission to ICU 30 days prior to IFI ( p=0.009). Aspergillosis was associated with reduced 12-week mortality ( p=0.001).On multivariate analysis age &amp;gt;40 ( p=0.010) and aspergillosis (p =0.002) remained independently associated with increased and reduced mortality, respectively. Conclusion This study reports unique information regarding proven and probable IFI in hematologic patients diagnosed under EORTC 2020 guidelines. Our epidemiology was similar to that reported by other countries, including most common etiologic agents and a higher proportion of AL and on active chemotherapy patients.Mortality rates were higher, possibly explained by the exclusion of possible IFI cases, higher proportion of active primary disease, and limited antifungal prophylaxis. Better survival among patients with aspergillosis, suggest that current diagnostic tests lead to earlier diagnosis and treatment with an impact on patients' prognosis.

1021. Semi-nested Molecular Method for the Diagnosis of Invasive Fungal Infections in Combat Injured

Abstract Background Among combat injured, invasive fungal infections (IFI) result in significant morbidity (eg surgical amputations). Primary diagnostic methods are limited as cultures are insensitive and time delayed, and histopathology often cannot provide genus level identification needed for directed treatment. We previously evaluated a panfungal PCR (PF) assay and it was 83% sensitive and 99% specific for angioinvasive IFI (AIFI). The PF assay requires substantial expertise and cannot be easily performed at all facilities. Thus, we evaluated less resource intensive semi-nested PCR (SN) targeted to clinically relevant fungi (order Mucorales and genus Aspergillus and Fusarium). Methods Formalin-fixed paraffin-embedded tissue (FFPE) specimens from a multicenter trauma IFI cohort (2009-14) were used. Cases were US military personnel injured in Afghanistan with histopathologic IFI evidence. Controls were patients with similar injury patterns and no laboratory IFI evidence (negative culture and histopathology). SN assays specific to Mucorales (V4/V5 regions of 18S rDNA), Aspergillus (P2/Asp2 regions of mitochondrial tRNA), and Fusarium (internal transcribed spacer [ITS]/28A regions of DNA) were compared to a PF assay amplifying the ITS2 region of rDNA and to histopathology. Performance characteristics and Cohen’s Kappa coefficients were calculated. Results Specimens from 92 injury sites (62 subjects) were compared to control specimens from 117 injuries (101 subjects; Fig 1). We observed substantial agreement between SN and PF assays overall and for order Mucorales (Table 1). Moderate agreement was observed at the genus level for Aspergillus and Fusarium. When compared with histopathology, sensitivity and specificity of SN assays were 64.7% and 96.6% respectively (91.7% and 96.6% when restricted to AIFI; Table 2). No statistically significant difference in performance between the SN and PF assays was observed. Conclusion Prior studies of SN molecular diagnostics have focused on culture-negative samples from immunocompromised patients. Our findings underscore utility of the SN approach in diagnosing IFI using FFPE samples, especially AIFI. Given the known DNA degradation with formalin fixation, sensitivity of SN assays may be higher in unfixed specimens and should be examined. Disclosures All Authors: No reported disclosures

Incidence of Invasive Fungal Infections in CAR T-Cell Therapy Recipients: A Single-Center Retrospective Analysis

Background: CAR T-cell therapy has transformed the treatment of hematologic malignancies, but challenges persist due to toxicities like cytokine release syndrome (CRS), immune effector cell-associated neurotoxicity syndrome (ICANS), and infections. Rates of invasive fungal infections (IFI) post CAR T-cell therapy are estimated to range from 1% to 7% (Haidar et al. Clin Infect Dis, 2020). Some centers use positive fungal biomarkers, such as serum (1à3)-β-D-glucan (B-DG) and galactomannan (GM), alongside surveillance radiographic imaging to guide preemptive fungal therapy in this population. The objective of the study was to assess the diagnostic performance of surveillance B-DG and GM for the diagnosis of IFI in recipients of BCMA and CD19-directed CAR T for multiple myeloma (MM) and non-Hodgkin lymphoma (NHL), respectively. Methods: We conducted a retrospective analysis on adult patients who received BCMA-directed CAR T therapy for MM or CD19-directed CAR T for NHL at our institution between 2018-2023. IFI was defined using the Revised European Organization for Research and Treatment of Cancer and the Mycoses Study Group (EORTC/MSG) criteria (Donnelly et al. Clin Infect Dis, 2020). Only patients with proven or probable infection were included in the analysis. The choice of antifungal prophylaxis was determined based on the treating physician's discretion. In the most cases, B-DG and GM monitoring was done prior to apheresis, before CAR T infusion, and subsequently every 14 days for a duration of 6 months. All patients were followed until 7/12/2023, or until their data were censored at the last follow-up. The cumulative incidence was calculated using Kaplan-Meier method, considering the time to IFI, with IFI-free death and disease relapse as competing risks. Statistical analysis was performed using IBM SPSS (version 29.0). Result: Among the 148 subjects in this study, 51 patients (34.5%) received BCMA CAR T, and 97 (65.5%) received CD19 CAR T therapy. The median age at CAR T infusion was 66 years (range 36-78) in the BCMA cohort and 65 years (range 27-83) in the CD19 cohort (p=0.64). The BCMA cohort had a higher number of prior therapies, with a median of 6 (range 2-15) compared to the CD19 cohort, which had a median of 4 (range 2-12) (p&amp;lt;0.001). The median follow-up from CAR T infusion was 9.5 months (range 0-63) in both cohorts, with 12 months (range 0-63) in the BCMA cohort and 8 months (range 0-49) in the CD19 cohort. All patients received prophylactic antifungal agents, including anti-yeast (fluconazole, n=95), and anti-mold (micafungin n=5, isavuconazole n=1, or Posaconazole n=52). Among the BCMA cohort, 92% of the patients received anti-yeast prophylaxis, while 8% were given anti-mold prophylaxis. In the CD19 cohort, 49.5% received anti-yeast prophylaxis, while 50.5% received anti-mold prophylaxis. Out of the 148 patients, 21 individuals (14%) exhibited elevated B-DG levels, while only 2 patients (1%) showed elevated GM levels. However, 20 out of 21 (95%) cases with positive B-DG, and all cases with positive GM were deemed to be false positives. Among these false positives, 60% were attributed to IVIG administration, while the cause for the remaining cases remained unknown (Table 1). Only one patient in this study developed IFI. The patient received anti-yeast prophylaxis (Fluconazole) at CAR-T and had a disseminated Rhizopus infection on day 13 post CD19 CAR T, which ultimately resulted in the patient's death. The 1-year cumulative incidence of IFI, considering competing risks, was 0.7% (95% confidence interval [CI], 0- 2.1). The median progression-free survival (PFS) in the overall cohort was 11.7 months (95% CI, 6.9-16.6), with 8.6 months (95% CI, 3.2-13.97) in the BCMA cohort and 14.6 months (95% CI, NR-NR) in the CD19 cohort (p=0.324). The median overall survival (OS) of the study cohort was 32.4 months (95% CI, 16.3-48.4), with 41.4 months (95% CI, 16.74-66.08) in the BCMA cohort and 23.98 months (95% CI, NR-NR) in the CD19 cohort (p=0.260). Conclusion: The rate of IFI in our cohort was low (0.7%) post-CAR T therapy. The diagnostic performance of serial monitoring of B-DG and GM for the diagnosis of IFI is poor in recipients of BCMA-directed CAR T therapy for MM or CD19-directed CAR T for NHL.