Anti-mold Prophylaxis Research Articles

The Incidence of Invasive Fungal Infections in Patients With AML Treated With a Hypomethylating Agent

BackgroundNewly diagnosed patients with acute myeloid leukemia (AML) who receive induction with a hypomethylating agent (HMA) are often neutropenic with an increased risk for invasive fungal infections (IFIs). This study analyzed the incidence and risk factors for IFIs in these patients, evaluated clinical patterns in antifungal prophylaxis, and assessed the diagnostic utility of tests in this setting. Patients and MethodsWe studied 117 newly diagnosed patients with AML treated with HMAs at our center, divided into groups based on concern for IFI (cIFI: all possible, probable, and proven IFIs) versus no concern for IFI. The Fisher exact test compared patients with cIFI versus without, and a multivariable logistic regression model estimated odds for cIFI. ResultsSixty-seven (57%) patients had cIFI, with 48 possible IFIs, 17 probable, and 2 proven cases. There was no difference in incidence based on home zip code, but the presence of chronic obstructive pulmonary disease was highly associated with cIFI (P = .001), as was male gender (P = .01). Neutropenia at treatment initiation was borderline in significance (P = .08). In diagnostics, 9% of patients had positive serum fungal markers, and 30 patients underwent bronchoscopy, with only 27% of cases yielding positive results. There was a difference in treatment regimens between patients receiving antifungal prophylaxis with mold coverage versus without mold coverage with respect to cIFI (P = .04). ConclusionscIFI in patients with AML treated with HMAs remains significant, especially in males and those with chronic obstructive pulmonary disease, who were found to be at higher risk. This may prompt clinicians to consider anti-mold prophylaxis in this setting.

Low Rate of Invasive Fungal Infections During Induction and Consolidation Chemotherapy for Adults with De Novo Acute Myeloid Leukemia Without Anti-mold Prophylaxis: Single-Center 2002-2018 Empirical/Pre-emptive Approach.

Broad-spectrum antifungal prophylaxis is currently considered the standard of care for adults with de novo AML for the prevention of invasive fungal infections (IFIs), especially invasive pulmonary aspergillosis (IPA). Because fluconazole has been used in our center as anti-yeast prophylaxis, we sought to analyze in detail the incidence of IFIs over a 17-year period, as well as their impact on outcome. A standardized protocol of patient management, including serum galactomannan screening and thoracic CT-guided diagnostic-driven antifungal therapy, was used in all patients. A total of 214 consecutive adults with de novo AML who were treated in 3 CETLAM (Grupo Cooperativo para el Estudio y Tratamiento de las Leucemias Agudas y Mielodisplasias) protocols from 2002 to 2018 were included. The 90-day incidence of any IFI (including possible cases) was 11% (95% CI 4-15%), most cases occurred during induction chemotherapy (8%, 95% CI 4-12%), and most cases were probable/proven IPA (8%, 95% CI 3-13%). Developing an IFI during induction and consolidation had no impact on 1-year survival. A case-control study with 23 cases of IPA and 69 controls identified induction/re-induction chemotherapy, chronic pulmonary disease and age > 60 years/poor baseline performance status as potential pretreatment risk factors. The current study proves that inpatient induction and consolidation chemotherapy for de novo AML can be given in areas with "a priori" high-burden of airborne molds with fluconazole prophylaxis, while the selective use of anti-mold prophylaxis in patients at very high risk may further reduce the incidence of IFI in this specific clinical scenario.

Invasive Aspergillosis Due to Aspergillus Section Usti: A Multicenter Retrospective Study.

Aspergillus spp. of section Usti (A. ustus) represent a rare cause of invasive aspergillosis (IA). This multicenter study describes the epidemiology and outcome of A. ustus infections. Patients with A. ustus isolated from any clinical specimen were retrospectively identified in 22 hospitals from 8 countries. When available, isolates were sent for species identification (BenA/CaM sequencing) and antifungal susceptibility testing. Additional cases were identified by review of the literature. Cases were classified as proven/probable IA or no infection, according to standard international criteria. Clinical report forms were obtained for 90 patients, of whom 27 had proven/probable IA. An additional 45 cases were identified from literature review for a total of 72 cases of proven/probable IA. Hematopoietic cell and solid-organ transplant recipients accounted for 47% and 33% cases, respectively. Only 8% patients were neutropenic at time of diagnosis. Ongoing antimold prophylaxis was present in 47% of cases. Pulmonary IA represented 67% of cases. Primary or secondary extrapulmonary sites of infection were observed in 46% of cases, with skin being affected in 28% of cases. Multiple antifungal drugs were used (consecutively or in combination) in 67% of cases. The 24-week mortality rate was 58%. A. calidoustus was the most frequent causal agent. Minimal inhibitory concentrations encompassing 90% isolates (MIC90) were 1, 8, >16, and 4 µg/mL for amphotericin B, voriconazole, posaconazole, and isavuconazole, respectively. Aspergillus ustus IA mainly occurred in nonneutropenic transplant patients and was frequently associated with extrapulmonary sites of infection. Mortality rate was high and optimal antifungal therapy remains to be defined.

Limited Positive Predictive Value of β-d-Glucan in Hematologic Patients Receiving Antimold Prophylaxis.

BackgroundDiagnostic value of β-d-glucan (BDG) in populations with low prevalence of invasive fungal infection (IFI), such as hematologic patients receiving antimold prophylaxis, should be re-evaluated.MethodsWe retrospectively reviewed episodes with BDG results in hematologic patients receiving antimold prophylaxis from January 2017 to August 2019 in a tertiary hospital. The episodes were classified as true positive ([TP] positive BDG with IFI), true negative ([TN] negative BDG without IFI), false positive ([FP] positive BDG without IFI), false negative ([FN] negative BDG with IFI), and nonevaluable.ResultsA total of 203 episodes were analyzed: 101 episodes (49.8%) were from stem cell transplants, 89 (43.8%) were from induction chemotherapy, and 13 (6.4%) were from graft-versus-host disease treatment. There were 62 nonevaluable episodes. Among 141 evaluable ones, there were 8 (5.7%) episodes of probable/proven IFI. True positive, TN, FP, and FN cases were 4 (2.8%), 112 (79.4%), 21 (14.9%), and 4 (2.8%) episodes, respectively. Sensitivity, specificity, positive predictive value, and negative predictive value were 50.0%, 84.2%, 16.1%, and 96.5%, respectively. Positive predictive value was 26.7% and 0.0% in diagnostic and surveillance episodes, respectively.Conclusionsβ-d-glucan test should be used to exclude IFI rather than for diagnosis in these patients.

1696. Epidemiology, Clinical Characteristics and Outcomes of Invasive Aspergillosis in a Tertiary Care Hospital in Mexico

BackgroundInvasive aspergillosis is an important cause of life-threatening infection in immunocompromised patients. The objective was to describe the epidemiology, clinical characteristics, and outcome of patients with invasive aspergillosis (IA) in a tertiary care center in Mexico.MethodsA laboratory-based survey was done to identify patients with positive Aspergillus culture or galactomannan from 2014 to 2018. The medical records were reviewed to include patients with proven and probable IA, according to the EORTC criteria. Descriptive analysis of clinical characteristics and risk factors for 6-week mortality was made through X2, T-test or Mann–Whitney test. A multivariate logistic regression model including variables with a P-value of <0.2 in univariate analysis was made.Results240 cases of IA were identified: 193 (80%) probable, 27 (11%) proven, and 20 (8.3%) not meeting the EORTC criteria but considered infection. 53% were male, median age was 44 years (IQR 28–58), 78 (32.5%) had acute leukemia (AL), 42 (17.5%) hematological neoplasia, 29 (12%) hematopoietic stem-cell transplant (HSCT), 25 (10.4%) solid-organ transplant and 44 (18.3%) autoimmune diseases, 17.5% patients with AL underwent induction remission chemotherapy of which 31% received antifungal prophylaxis. Among patients with IA, 183 (82%) had a positive galactomannan and 109 (45%) had a culture with Aspergillus. Eleven had > 1 species: 55/120 (46%) were A. fumigatus, 18 (15%) A. niger and 18 (15%) A. flavus. Pulmonary disease occurred in 214 (89%). 212 patients (88%) received antifungal treatment with a median duration of 42 days (IQR 20–42). 129 (61%) received voriconazole (VRC), 20 (8.3%) Amphotericin B and 20(8.3%) were randomized to a posaconazole vs. VRC trial. Six-week mortality was 35% (n = 85). Lymphopenia (OR 3.6; 95% CI 1.4–9.0), liver failure (OR 3.3; 95% CI 1.7–6.5) and older age (OR 1.03; 95% CI 1.01–1.05) (marginally) were independently associated with increased 6-week mortality.Conclusion240 patients with IA were identified in a 5-year period in a tertiary care center. Most had hematological neoplasias and low prevalence of antimold prophylaxis due to economical reasons. Six-week mortality was 35%, nonsurvivors had liver failure and lymphopenia more often. Increased awareness to prevent IA is needed. Disclosures All authors: No reported disclosures.

273. Low Positive Predictive Value of β-d-Glucan in Hematology Patients Receiving Antimold Prophylaxis

BackgroundDetection of β-D-glucan (BDG) in serum is recognized as the mycological evidence in the diagnosis of invasive fungal infection (IFI). However, its diagnostic value in low prevalence of IFI has not been elucidated. We aimed to examine the performance of BDG in hematology patients receiving antimold prophylaxis.MethodsWe retrospectively reviewed all BDG results performed for the purpose of diagnosis or surveillance for IFI in hematology patients receiving posaconazole or micafungin prophylaxis from January 2017 to February 2019 in a tertiary hospital. At least two consecutive positive results of BDG were regarded as positive BDG. All the episodes were classified into true-positive (TP, positive BDG with probable/proven IFI), true-negative (TN, negative BDG without probable/proven IFI), false-positive (FP, positive BDG without probable/proven IFI), false-negative (FN, negative BDG with probable/proven IFI), and nonevaluable (could not be determined for the occurrence of breakthrough IFI). When BDG test was performed in the setting of persistent fever ≥72 hours in spite of broad-spectrum antibiotics or with a suspicion of IFI, it was defined as a diagnostic BDG episode, while others were defined as a surveillance BDG episode.ResultsOf a total of 140 episodes, 24 episodes were non-evaluable. Among 116 evaluable episodes, 75 received induction chemotherapy for acute leukemia or myelodysplastic syndrome, 35 underwent stem cell transplantation, and 10 had intensive treatment for graft-vs.-host disease. There were three episodes of probable/proven IFI (2.6%). Ninety-one (78.4%) were performed with diagnostic purpose, while 25 (21.6%) were performed for surveillance. TP, TN, FP, and FN were 2 (1.7%), 91, 22, and 1, respectively. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value were 66.7%, 80.5%, 8.3% and 98.9%, respectively. PPV was 13.3% and 0% in diagnostic and surveillance BDG episodes, respectively.ConclusionThe PPV of BDG was low in hematology patients receiving antimold prophylaxis, even in the diagnostic-driven episodes. The routine screening of BDG is not helpful, and the BDG test may be used for exclusion of IFI rather than for diagnosis in these patients. Disclosures All authors: No reported disclosures.

Invasive pulmonary aspergillosis in acute leukemia: a still frequent condition with a negative impact on the overall treatment outcome

We evaluated the impact of invasive pulmonary aspergillosis (IPA) on epidemiology and outcome in acute leukemia (AL), analyzing all acute myeloid (AML) and acute lymphoblastic leukemia (ALL) consecutively admitted to our Institution during a 5-year period of observation. Only AML patients received anti-mold prophylaxis. Among 175 AL patients (136 AML/39 ALL), possible and proven/probable IPA were diagnosed in 28 (16%). Frequency of IPA was similar in AML (16.2%) and in ALL (15.4%). Two-year overall survival (OS) was significantly affected by IPA (no IPA: 69.8% vs IPA: 31.7% p = .002). OS was similar in patients with proven/probable (28.2%) and possible IPA (36.4%) (p = .003 and .065, respectively). When censoring patients at transplant, IPA still affected 2-year survival (49.6% vs 79.2%, p = .02), but only proven/probable IPA was associated with lower survival (34.7%, p = .0003). IPA negatively impacts on long-term survival of leukemia patients; antifungal prophylaxis should be adopted also during induction in ALL and in AML beyond induction therapy.

1131. Characteristics and Risk Factors for Mortality in Hematologic Patients with Invasive Non-Aspergillus Mold Infections: A Single-Center 7-Year Cohort Study

BackgroundSince anti-mold active azole prophylaxis has become a standard approach for patients with high-risk hematologic diseases, the epidemiology of invasive fungal infections (IFIs) has shifted toward non-Aspergillus molds in recent years. The aim of this study was to identify the epidemiology and characteristics of non-Aspergillus invasive mold infections (NAIMIs) in the real-world.MethodsProven/probable NAIMIs developed in patients with hematologic diseases were reviewed from January 2011 to January 2018 at the Catholic Blood and Marrow Transplantation Center.ResultsThere were 662 patients with proven/probable IMIs, of which 40 patients (41 isolates) were diagnosed with NAIMIs. The incidence of NAIMIs showed an increasing trend since 2013 when posaconazole prophylaxis was approved in Korea [correlation coefficient (r) = 0.735, P = 0.265]. Mucormycosis (n = 24, 58.5%) was the most common, followed by Fusarium (n = 7, 17.1%), Alternaria (n = 2), Scopulariopsis (n = 2), Scedosporium (n = 2), Paecilomyces (n = 1), Coprinus (n = 1), Chaetomium (n = 1), and Schizophyllum (n = 1). Twenty-eight patients were under neutropenia upon diagnosis of NAIMI and 35.0% were allogeneic stem cell transplantation recipients. The most common sites of NAIMI were the lungs (60.0%), followed by the paranasal sinus (17.5%) and disseminated infections (12.5%). There were 35.5% breakthrough IMI cases. In addition, there were 42.5% mixed or concurrent IFIs and 77.5% had coexisting bacterial or viral infections. The overall mortality at 6 and 12 weeks was 32.5% and 42.5%, respectively. The mortality rates for mucormycosis and nonmucormycosis at 6 weeks were 21.7% and 47.1%, respectively. Breakthrough IFIs [adjusted hazards ratio (HR) = 4.83, P = 0.018] and surgical treatment [HR = 0.09, P = 0.003] were independently associated with 6-week mortality.ConclusionNAIMIs showed an increasing trend, mixed/concurrent IFIs were substantial, and coexisting bacterial or viral infections were found in more than two-thirds of patients. Breakthrough IFIs and surgical treatment have significant impact on mortality. More meticulous approaches to diagnosis and treatment strategies of NAIMIs are needed.Disclosures All authors: No reported disclosures.

Sequential systematic anti-mold prophylaxis with micafungin and voriconazole results in very low incidence of invasive mold infections in patients undergoing allogeneic hematopoietic stem cell transplantation.

Recipients of allogeneic hematopoietic stem cell transplantation (allo-HSCT) are at high risk for invasive mold infections (IMI). The goal of the study is to describe the incidence and outcome of IMI in patients after allo-HSCT in a large cohort of patients receiving anti-mold prophylaxis. We conducted a retrospective review of 988 consecutive adults who underwent allo-HSCT in our center from 2008 through 2014. Standard prophylaxis consisted of micafungin 150mg IV daily from admission to day +7±3 followed by voriconazole until day +75 to +100. Cases meeting criteria for proven or probable IMI according to EORTC-MSG criteria were included. Median age at HSCT was 54years. The most common diagnoses were acute myeloid leukemia (n=351, 36%) and lymphoid malignancies (n=248, 25%). Matched related or unrelated donors (URD) were used in 686 (69%) patients, mismatched URD in 142 (14%) and cord blood units in 154 (16%). Twenty-one patients were diagnosed with IMI after allo-HSCT, 19 probable and 2 proven, and one patient was diagnosed postmortem. Microbiological diagnosis was established in 9 cases, 5 of them being Aspergillus. One-year cumulative incidence (CI) of IMI was 1.6% (95% CI 0.9-2.5) while 12-week overall survival after IMI was 39% (95% CI 24-65) Analyzed by disease, there was a trend for a higher 1-year CI of IMI in patients with ALL (5% [95% CI 1.6-11.4]) when compared with AML (1.4%), MDS (1.5%) and lymphoma (1.2%), P=.06. The 1-year CI of IMI after transplantation is low in patients receiving anti-mold prophylaxis with micafungin bridged to voriconazole, although these infections are associated with a higher risk of mortality.

Spotlight on isavuconazole in the treatment of invasive aspergillosis and mucormycosis: design, development, and place in therapy.

In recent decades, important advances have been made in the diagnosis and treatment of invasive aspergillosis (IA) and mucormycosis. One of these advances has been the introduction of isavuconazole, a second-generation broad spectrum triazole with a favorable pharmacokinetic and safety profile and few drug–drug interactions. Phase III trials in patients with IA and mucormycosis demonstrated that isavuconazole has similar efficacy to voriconazole for the treatment of IA (SECURE trial) and liposomal amphotericin B for the treatment of mucormycosis (VITAL trial with subsequent case–control analysis) and a favorable safety profile with significantly fewer ocular, hepatobiliary, and skin and soft tissue adverse events compared to voriconazole. As a result, recent IA guidelines recommend isavuconazole (together with voriconazole) as gold standard treatment for IA in patients with underlying hematological malignancies. In contrast to liposomal amphotericin B, isavuconazole can be safely administered in patients with reduced renal function and is frequently used for the treatment of mucormycosis in patients with reduced renal function. Updated guidelines on mucormycosis are needed to reflect the current evidence and give guidance on the use of isavuconazole for mucormycosis. Studies are needed to evaluate the role of isavuconazole for 1) anti-mold prophylaxis in high-risk patients, 2) salvage treatment for IA and mucormycosis, and 3) treatment for other mold infections such as Scedosporium apiospermum.

Challenges in microbiological diagnosis of invasive Aspergillus infections

Invasive aspergillosis (IA) has been increasingly reported in populations other than the historical hematology patients and there are new questions about the performance of microbiological tools. Microscopy and culture have been completed by biomarkers, either antigens or DNA, and in blood or respiratory specimens or both. First studied in hematology, the antigen galactomannan performance in serum is low in other patient populations where the pathophysiology of the infection can be different and the prevalence of IA is much lower. DNA detection with polymerase chain reaction (PCR) in blood or serum (or both) has reached a certain level of acceptance thanks to consensus methods based on real-time quantitative PCR (qPCR). When used on respiratory specimens, galactomannan and qPCR depend on standardization of the sampling and the diverse mycological procedures. Thus, culture remains the main diagnostic criterion in critically ill patients. The current trend toward more effective anti-mold prophylaxis in hematology hampers the yield of a screening strategy, as is usually performed in hematology. Therefore, circulating biomarkers as confirmatory tests should be considered and their performance should be reappraised in each new setting. The use of azole prophylaxis also raises the issue of selecting azole-resistance Aspergillus fumigatus isolates. Ideally, the biomarkers will be more efficient when individual genetic risks of IA are defined. Culture, though not standardized, remains a key element for the diagnosis of IA and has the advantage to easily detect molds other than A. fumigatus. It is still unclear whether next-generation sequencing will replace culture in the future.

Serum galactomannan surveillance may be safely withdrawn from antifungal management of hematology patients on effective antimold prophylaxis: a pilot single-center study.

Serum galactomannan surveillance may be safely withdrawn from antifungal management of hematology patients on effective antimold prophylaxis: a pilot single-center study

Antimold Prophylaxis May Reduce the Risk of Invasive Fusariosis in Hematologic Patients with Superficial Skin Lesions with Positive Culture for Fusarium.

Hematologic patients with superficial skin lesions on admission growing Fusarium spp. are at a high risk for developing invasive fusariosis during neutropenia. We evaluated the impact of primary prophylaxis with a mold-active azole in preventing invasive fusariosis in these patients. Between August 2008 and December 2014, patients with acute leukemia or aplastic anemia and recipients of hematopoietic cell transplants were screened on admission with dermatologic and direct exams and fungal cultures of superficial skin lesions. Until November 2009, no interventions were made. Beginning in December 2009, patients with baseline skin lesions and a direct exam and/or culture suggestive of the presence of Fusarium spp. received prophylaxis with voriconazole or posaconazole. Skin lesions in the extremities (mostly onychomycosis and interdigital intertrigo) were present on admission in 88 of 239 episodes (36.8%); 44 lesions had hyaline septate hyphae identified by direct exam, and cultures from 11 lesions grew Fusarium spp. Antimold prophylaxis was given for 20 episodes (voriconazole for 17 and posaconazole for 3). Invasive fusariosis was diagnosed in 14 episodes (5.8%). Among patients with baseline skin lesions with positive cultures for Fusarium spp., 4 of 5 without antimold prophylaxis developed invasive fusariosis versus 0 of 6 with antimold prophylaxis (P = 0.01; 95% confidence interval for the difference between proportions, 22% to 96%). Primary antifungal prophylaxis with an antimold azole may prevent the occurrence of invasive fusariosis in high-risk hematologic patients with superficial skin lesions on admission growing Fusarium spp.

Risk Factors Associated With Early Invasive Pulmonary Aspergillosis in Kidney Transplant Recipients: Results From a Multinational Matched Case-Control Study.

Risk factors for invasive pulmonary aspergillosis (IPA) after kidney transplantation have been poorly explored. We performed a multinational case-control study that included 51 kidney transplant (KT) recipients diagnosed with early (first 180 posttransplant days) IPA at 19 institutions between 2000 and 2013. Control recipients were matched (1:1 ratio) by center and date of transplantation. Overall mortality among cases was 60.8%, and 25.0% of living recipients experienced graft loss. Pretransplant diagnosis of chronic pulmonary obstructive disease (COPD; odds ratio [OR]: 9.96; 95% confidence interval [CI]: 1.09-90.58; p = 0.041) and delayed graft function (OR: 3.40; 95% CI: 1.08-10.73; p = 0.037) were identified as independent risk factors for IPA among those variables already available in the immediate peritransplant period. The development of bloodstream infection (OR: 18.76; 95% CI: 1.04-339.37; p = 0.047) and acute graft rejection (OR: 40.73, 95% CI: 3.63-456.98; p = 0.003) within the 3 mo prior to the diagnosis of IPA acted as risk factors during the subsequent period. In conclusion, pretransplant COPD, impaired graft function and the occurrence of serious posttransplant infections may be useful to identify KT recipients at the highest risk of early IPA. Future studies should explore the potential benefit of antimold prophylaxis in this group.

Systematic Anti-Mold Prophylaxis Results in Very Low Incidence of Invasive Fungal Infections in Patients Undergoing Allogeneic Hematopoietic Cell Transplantation

Recipients of Cord blood (CBT) T-cell depleted (TCD) and mismatched unrelated (URD) donors allografts are at high risk for invasive mold infections (IMI). The impact of mold-active prophylaxis (PPX) on the incidence and outcomes of mold infections is not well defined. Retrospective review of 988 consecutive adults who underwent alloHCT in our center from 2008 through 2014. Standard PPX consisted of micafungin 150mg IV daily from admission to Day (D) +7 post HCT followed by voriconazole through at least D+100. PPX was continued beyond D+100 in pt with GVHD or use of corticosteroids. Patients (pts) with intolerance or contraindication to voriconazole received posaconazole or micafungin. Pts with IMI were identified through microbiology and pathology records. Cases meeting criteria for proven or probable according to EORTC-MSG criteria are included in the analysis. Median age at HCT was 54 years (range 18-76). The most common diagnoses were AML (n = 351, 36%) and lymphoma (n = 214, 21%). Matched-related and URD were used in 686 (69%) pts, mismatched URD and CBT in 142 (14%) and 154 (16%) pts, respectively. Ex-vivo TCD was used in 484 (49%) pts. 63% pts received myeloablative conditioning. Twenty-four pts (2.4%) developed IMI at a median of 203 days (range 7-1327) after HCT. 19 (79%) cases were probable and 5 (21%) proven. Microbiological diagnosis was established in 10 cases (aspergillus spp=5, mucor=5). Thirteen (54%) pts were diagnosed with probable IFI by positive Beta-D-Glucan assay, compatible radiological findings and host factors. Pneumonia was the most presentation (n = 21, 88%). Fourteen (58%) pts received systemic corticosteroids within 30 days prior to diagnosis, including 12 for GVHD. 14 of the 24 pts (58%) were not on the planned anti-mold prophylaxis at the time of IFI, due to azole-related toxicity (n = 7), drug allergy (n = 1) or patient preference (n = 1). 5 pts were off prophylaxis as they were beyond day +100 and had no additional risk factors. No risk factors for development of IFI were identified, due to low number of events. Subgroup analysis identified HCT populations with very low risk of IFI including AML in CR1 (1/248, 0.4%) and myeloproliferative neoplasms (0/58), and others with higher risk such as CBT (5/154, 3.2%) and mismatched URD (6/142, 4.2%). In the TCD-PBSC setting, 12 of 484 pts (2.5%) developed an IFI. The 12 weeks and 1 year overall survival after IFI was 42% (95%CI 32-52) and 29% (95%CI 19-38), respectively (Figure). All 5 pts with Mucor died at a median of 18 days (1-146) of diagnosis (4/5 related to IMI). 1) In this large and heterogeneous cohort including pts at high risk for mold infection who received mold active prophylaxis, we observed low incidence (2.4%). 2) IMI associated mortality remained high, highlighting the need for improved diagnostic techniques and therapeutic strategies.

Testing the Performance of Lateral Flow Assay (LFA) for the Diagnosis (Dx) of Invasive Pulmonary Aspergillosis (IPA) in the Era of Anti-mold Prophylaxis

Testing the Performance of Lateral Flow Assay (LFA) for the Diagnosis (Dx) of Invasive Pulmonary Aspergillosis (IPA) in the Era of Anti-mold Prophylaxis

Screening Efectiveness with SERUM Galactomannan in High and Low Risk Patients for Fungal Infection in the Post ALLO-HTC

Background and aims: Serum galactomannan (GM) is used as a screening test for the early diagnosis of Aspergillus infection in high risk patients for fungal invasive infection. Serial GM levels analysis have proven to be useful in the high risk clinical setting patients, specially when neutropenic and not receiving anti-mold agents prophylaxis. There is a lack of information regarding the benefit of GM in patients undergoing allogeneic hematopoietic cell transplant (HCT). The aim of this work is to measure the real clinical benefit of the serial periodic determination of GM in the post allo-HCT period.Material and methods: 139 consecutive patients who receivedan allo-HCT (59% related family member donorsand 41% unrelated donors) in our centre for five years (since January 2010 to February 2015) were included in the study. Median age was 46 years. Baseline characteristics of these patients are shown in figure 1. Patients were monitored with GM weekly and received primary prophylaxis with fluconazole since the admission until the immunosuppression was tapered.In order to find a population that could benefit the most for AGA monitoring, we classified our population in low risk patients for invasive fungal infection (IFI) versus high risk patients (those with previous proven or probable IFI or those suffering from GVHD; high risk patients received anti-mold prophyllaxis, mainly with voriconazole or posaconazole). Patients considered as low risk who suffered from Graft versus Host Disease (GVHD) in the ulterior outcome, were censored for low risk and considered as high risk since the development of GVHD, and therefore anti-mold agent prophylaxis was started. GM positivity was determined according standard criteria. When GM positivity was detected, radiological and clinical studies (chest/sinus CT scans, cultures, etc.) to discard Aspegillosis were done as soon as possible. Every patient was followed up prospectively until the last medical consultation or decease.Results: Global overall survival (OS) for the entire cohort was 55.39% and cumulative incidence for severe GVHD grade III/IV was 49.5%. During the follow up, GM became positive in 31/139 (22%) cases. With this approach, the global false positive and false negative rate was 31% and 6% respectively.110/139 (79.14%) patients were identificated as low risk cases. We observed GM positivization in 1.81% (2/110) and 37.18% (29/78) for low and high group respectively. All 2 positive GM in the low risk group were false positives. Regarding the high risk group, 34.48% (10/29) were false positives while in the rest 19 patients (65.52%), subsequent radiological and clinical findings allowed us to diagnose Aspergillus infection (besides they received anti-mold agent prophylaxis).Conclusions: In our experience there is not enough evidence for supporting making serial monitoring with GM in low risk patients for IFI in the post allo-HCT period. However it may be an useful tool in high risk patients.Table 1N (%)Age (years)Mean 46.18 Median 48.01Sex (man vs woman)91 vs 48 (65 vs 35%)Hematology disease Acute Mieloid Leukemia Acute Limphoblastic Leukemia Multiple Myeloma Chronic Mieloid Leukemia Non Hodgkin Lymphoma Hodgkin Lymphoma Others diseases79 (56.8%) 18 (12.9%) 9 (6.5%) 3 (2.2%) 13 (9.4%) 9 (6.5%) 8 (5.6%)Pre allo-HSCT IFI17 (12.2%)Aconditioning Myeloablative Reduced Intensity95 (68.3%) 44 (31.7%)Source of progenitors Peripherical blood Bone marrow132 (95.0%) 7 (5.0%)Type of donor Related Non-related82 (59.0%) 57 (41.0%)Graft time (days) Neutrophils (>500 in two consecutive determinations) Platelets (>30.000 in two consecutive determinations)15.52 17.75Post-transplant CMV viremia (number of patients with >600 copies in the post-HSCT period)48 (34.5%) DisclosuresNo relevant conflicts of interest to declare.

Whole Genome Sequencing of Azole-Resistant Aspergillus Fumigatus Strains from Hematopoietic Stem Cell Recipients Identifies Candidate Molecular Targets Potentially Implicated in Novel Resistance Mediating Mechanisms - First Results

IntroductionInvasive aspergillosis (IA) represents a critical complication that frequently emerges during induction chemotherapy for acute leukemia or after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and is associated with high mortality rates. During the last years, routine administration of antimold prophylaxis based on triazole regimens provided a major advance in controlling fungal infections in patients at high risk for IA. However, several studies recently reported azole resistant fungal strains, which are detectable in patients at risk with rapidly increasing frequency. While mutations in the 14α-sterol demethylase (cyp51A) gene have been implicated in mediating azole resistance, in approximately 50% of resistant patient isolates the underlying genetic cause is unknown (van der Linden, Emerging Infectious Diseases, 2015). In view of a rising prevalence of azole resistance in the high risk clinical setting we performed whole genome sequencing (WGS) of azole-resistant cyp51A-wildtype Aspergillus fumigatus (Afu) specimen, the main effector of IA, with the aim to identify novel genetic alterations leading to azole resistance.MethodsAfu clinical strains were isolated from six different patients with IA undergoing intensive chemotherapy for acute leukemia or after allo-HSCT. Phenotypic resistance screening confirmed azole resistance in all samples. Moreover, Sanger sequencing revealed characteristic resistance associated cyp51A mutations ("TR46/Y121F/T289A") in four of the six specimen, while two samples harbored cyp51A wildtype sequences. High molecular weight genomic DNA (gDNA) was isolated using a modified DNeasy Plant Mini Kit protocol (Qiagen). WGS sequencing libraries were prepared utilizing the transposome fragmentation based Nextera XT DNA Library Prep Kit (Illumina) and sequenced on a MiSeq next generation sequencing (NGS) system (Illumina) with V2 sequencing chemistry (2x250bp). Raw paired reads were mapped to the NCBI Af293 (ASM265v1) reference genome using BWA followed by calibration with GATK. Subsequently, genomic variants were identified by GATK Haplotypecaller module and ANNOVAR software was used for functional variant annotation.ResultsWGS of four cyp51A mutated and two cyp51A wildtype samples resulted in an average genomic coverage of 26x (range 21.2x - 30.1x) allowing robust detection of genomic sequence variants. Of note, the presence or absence of cyp51A "TR46/Y121F/T289A" mutational genotype was reliably validated by WGS data using both our variant calling pipeline and manual screening of mapped reads with the IGV browser. In an attempt to screen for novel, previously undocumented candidate genes that are mechanistically involved in azole resistance we first identified all mutations that are exclusively present in the cyp51A wildtype samples and undetectable in the cyp51A mutated cohort. Next, we excluded all variants that have previously been reported as being polymorphic in WGS data from 22 different Afu strains sequenced by the J. Craig Venter Institute. Using this strategy we identified a core candidate cluster of 112 nucleotide variants in 62 genes that were exclusively found in the cyp51A wildtype group and predicted to have an impact on the amino acid sequence of the corresponding translational products. In particular, exemplary identified mutations affected genes coding for glycosyl hydrolase family 31 proteinimplicated in carbohydrate metabolic processes and 3-oxo-5-alpha-steroid 4-dehydrogenase family protein, which is, similar to cyp51A, involved in steroid metabolism.ConclusionThe rapid increase in detection of triazole resistant fungal strains in clinical contexts represents a critical problem, particularly for immunocompromised patients at high risk for IA, and highlights the need to explore the underlying molecular causes. Pending further validation in a larger cohort, our NGS-based approach for sequencing the genomes of azole resistant Afu strains identified several candidate genomic alterations that might pinpoint alternative pathways to acquisition of resistance, independent of the previously described cyp51A mutations. Ultimately, our approach for discovery of previously undescribed resistance pathways in fungal strains might aid in exploration of novel therapeutic targets and identify suitable biomarkers for effective clinical and epidemiological surveillance. DisclosuresBuchheidt:Gilead: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Pfizer: Other: travel grant, Research Funding, Speakers Bureau; Astellas: Other: travel grant, Speakers Bureau; Merck Sharp & Dohme: Other: travel grant, Speakers Bureau.

Correlation of Pain and Fluoride Concentration in Allogeneic Hematopoietic Stem Cell Transplant Recipients on Voriconazole

Supportive care guidelines recommend antimold prophylaxis in hematopoietic stem cell transplant (HSCT) recipients deemed to have high risk for invasive fungal infection, leading to long-term use of voriconazole after allogeneic HSCT in patients who remain immunocompromised. Voriconazole has been associated with periostitis, exostoses, and fluoride excess in patients after solid organ transplantation, HSCT, and leukemia therapy. The aims of this study were to describe the frequency and clinical presentation of patients presenting with pain and fluoride excess among allogeneic HSCT patients taking voriconazole, to identify when a plasma fluoride concentration was measured with respect to voriconazole initiation and onset of pain, and to describe the outcomes of patients with fluoride excess in the setting of HSCT. A retrospective review was conducted of all adult allogeneic HSCT patients receiving voriconazole at Mayo Clinic in Rochester, Minnesota, between January 1, 2009 and July 31, 2012. Of 242 patients included, 32 had plasma fluoride measured to explore the etiology of musculoskeletal pain. In 31 patients with fluoride measurement while on voriconazole, 29 (93.5%) had elevated levels. The median plasma fluoride was 11.1 μmol/L (range, 2.4 to 24.7). The median duration of voriconazole was 163 days (range, 2 to 1327). The median time to fluoride measurement was 128 days after voriconazole initiation (range, 28 to 692). At 1 year after the start of voriconazole after HSCT, 15.3% of patients had developed pain associated with voriconazole use and 35.7% developed pain while on voriconazole after 2 years. Of the patients with an elevated fluoride level, 22 discontinued voriconazole; pain resolved or improved in 15, stabilized in 3, and worsened in 4 patients. Ten patients continued voriconazole; pain resolved or improved in 7, was attributable to alternative causes in 2, and undefined in 1. Serum creatinine, estimated glomerular filtration rate, alkaline phosphatase, and voriconazole concentration did not predict for fluoride excess and associated pain. Periostitis due to fluoride excess is a common adverse effect of voriconazole that should be considered in patients presenting with pain and is often reversible after drug discontinuation. Alternative antifungal agents with a lower risk for fluoride excess should be considered in patients receiving voriconazole who develop fluoride excess and pain.

Invasive Fungal Infections in Acute Promyelocytic Leukemia Patients. Results of a Prospective Multicenter Study in Italy

OBJECTIVESAim of this prospective study was to evaluate the risk of invasive fungal infection (IFI) in patients (pts) with acute promyelocytic leukemia (APL) and to compare APL pts with patients affected by non promyelocytic acute myeloid leukemia (npAML) in order to evaluate factors potentially linked to IFI in these two subsets of acute myeloid leukemia.PATIENTS AND METHODSFrom January 2010 to April 2012 all pts with newly diagnosed AML were registered in 33 Italian participating centers. A minimum follow up of 90 days after 1st induction chemotherapy was requested for all pts. A prolonged follow up until June 2014 was made only for APL. Data were collected about age, gender, AML subtype, treatment and also about post chemotherapy risk factors for IFI (duration of neutropenia, mucosal damages, vomiting, diarrhea, presence of medical devices), antifungal prophylaxis, onset of IFI, level of certainty (possible/probable/proven), and antifungal treatment. Only for APL the survey was prolonged for at least 3 months in order to analyze if these pts have an IFI risk during other than first induction phases.RESULTS1,192 consecutive newly diagnosed adult AML pts (npAML:1,086/APL:106) were enrolled in the study.Among npAML pts, those receiving low dose chemotherapy and/or palliative treatment were excluded from the analysis; in the remaining 881 pts 214 cases (24%) of IFI were recorded.Considering APL, 3 pts were excluded from the analysis due to early death (1 pt) or bad performance status (2 pts). The remaining 103 pts received APL treatment according to local protocols: all trans retinoic acid (ATRA) plus chemotherapy (90 pts) or ATRA plus arsenic trioxide (ATO)(13 pts). Only 8 (8%) APL pts developed an IFI after the induction phase: 1 proven, 3 probable and 4 possible IFI. All cases were caused by molds. All APL were followed for a median follow up of 36 months (range 3-54). During this time only 2 other cases of IFI were observed: 1 possible IFI during consolidation at 16 weeks from APL diagnosis and 1 probable aspergillosis in a rare case of APL relapse at 132 weeks from APL diagnosis. All the IFI occurred in pts treated with ATRA plus chemotherapy.IFI was fatal in only 1 case (cerebral aspergillosis), all the other pts recovered after antifungal treatment.A comparison between npAML and APL was made in order to analyze the risk of IFI within 90 days after induction treatment among these 2 groups of patients (see table). A significantly lower number of overall IFI and systemic antifungal treatment was observed in the APL group, in spite of the fact that systemic anti mold prophylaxis was significantly less frequently utilized. Table 1Comparison between APL and npAML in induction phaseAPLnpAMLpNumber of pts103881Mean age51550.01m/f50/53448/433N.S.Performance status (WHO)0-1>1. 76 27. 284 597. <0.0001Central venous catheter52 (50%)687 (78%)<0.0001Neutropenia (<1000/mm3)103 (100%)874 (99%)N.S.Mean duration of neutropenia (<1000/mm3)23 days25 days0.1Mean duration of deep neutropenia (<500/mm3)17.5 days24 days0.04Antifungal prophylaxis94 (91%)837 (95%)N.S.Topical antifungal prophylaxis17 (17%)60 (7%)0.0005Drug in prophylaxisfluconazoleitraconazoleposaconazoleother.33 (32%)13 (12%)38 (37%)1 (1%).168 (19%)117 (13%)513 (58%)23 (3%).0.002N.S. <0.0001IFIsall casesproven/probable.8 (8%)4 (4%).214 (24%)77 (9%).0.00010.08moldsall casesproven/probable.8 (8%)4 (4%).191 (22%)55 (6%).0.0006N.S.yeastsall cases.0.23 (3%). <0.0001Antifungal treatmentMean duration11 (11%)17 days275 (31%)14 days<0.0001 N.S.Overall mortality at 30 days8 (8%)110 (12%)N.S.Mortality due to IFI at 30 days1 (1%)25 (3%)N.S.Comparing APL among them in order to identify parameters that could be correlated to IFI presentation, no significant factors were identified.DISCUSSIONIn our prospective study we specifically analyzed the incidence and the type of IFI in APL during a prolonged follow-up. Only 10 cases of IFI were documented and in most cases (6 pts) the infection was only possible. Comparing APL to npAML a lower incidence of overall IFI was observed despite less use of mold active drugs as prophylaxis. It could be attributed to the different chemotherapy (less aggressive in APL) and to lower duration of deep neutropenia. No yeast infection was observed in APL. On the basis of this study, APLs may be considered at low risk of IFI so probably the use of a mold active antifungal prophylaxis could be omitted. DisclosuresNo relevant conflicts of interest to declare.