Bronchoalveolar Lavage Galactomannan Research Articles

Comparison of Sensitivity of Low-Dose CT Scan and Serum Galactomannan in Patients with Hematologic Malignancies and Positive Bronchoalveolar Lavage for Invasive Pulmonary Aspergillosis

Abstract 1490▪▪This icon denotes a clinically relevant abstract Background:Invasive pulmonary aspergillosis (IPA) is associated with considerable morbidity and mortality in patients with acute leukemia and those undergoing hematopoietic stem cell transplantation (HSCT). Timely diagnosis of IPA is crucial, but difficult. Strategies for early detection of IPA include serum galactomannan (GM), bronchoalveolar lavage (BAL) GM and characteristic radiologic abnormalities on CT scan; however, the relative sensitivity of these diagnostic tests is unclear. We sought to determine the utility of serum GM and characteristic CT radiologic abnormalities in diagnosing IPA in patients with hematologic malignancies who had a positive BAL for Aspergillus. Methods:We performed a single center retrospective cohort study from 2010 to 2012 to determine the sensitivity serum GM (OD > 0.5) and low dose CT scan in patients with hematologic malignancies. All positive BAL GM samples and Aspergillus isolates from BAL fluid cultures from patients with acute leukemia and those undergoing HSCT were reviewed; all patients had abnormal chest CT scans consistent with pulmonary infection. IPA was classified as proven, probable or possible according to EORTC/MSG criteria. Recent low dose CT scans correlating to the date of the positive BAL GM or Aspergillus isolation were reviewed and graded: halo signs, cavities, crescents and nodules were deemed to be consistent with IPA, while other changes (e.g. ground-glass changes, consolidation) were considered non-specific. Results:A total of 49 BAL samples were included; of these, 31 were considered probable IPA and 18 possible IPA (on the basis of non-specific radiologic findings). Most patients had either no prior or 1–2 days of mold-active antifungal agents. There were 43 cases with positive BAL GM and 11 cases of positive Aspergillus BAL isolates; 5 patients had both. Of the positive BAL GM cases, 27 (63%) were associated with radiologic findings consistent with IPA. The remaining cases were associated with non-specific radiologic findings. Of the patients with Aspergillus isolates on BAL, 55% (6/11) had radiologic features consistent with IPA, while the remaining cases had non-specific radiologic findings.We next evaluated the sensitivity of serum GM. Of 34 patients with BAL GM positivity who had concomitant serum GM testing, 4 (12%) had positive serum GM. Of 8 patients with Aspergillus species isolated on BAL who had serum GM performed, only 1 (12.5%) had positive serum GM. In contrast, 5/8 patients (63%) with Aspergillus isolates on BAL had a positive BAL GM. The combined sensitivity of serum GM was 12% (5/42). Conclusions:Although the majority of patients with positive BAL for Aspergillus (+GM and/or isolates) had characteristic radiologic findings on CT scan, the absence of such findings did not exclude this diagnosis, as over one third had only non-specific radiologic findings. Serum GM had very low sensitivity in this population and should not be used in isolation to diagnose IPA. Disclosures:No relevant conflicts of interest to declare.

Diagnosis of invasive pulmonary aspergillosis in children with bronchoalveolar lavage galactomannan

Invasive pulmonary aspergillosis (IPA) is a life-threatening complication in immunocompromised patients. Early diagnosis and therapy improves outcome. Assessment of galactomannan (GM) in bronchoalveolar lavage (BAL) fluid is a proposed tool to diagnose IPA. Little is known about the diagnostic value of BAL GM in children. Retrospectively, 72 bronchoscopies were analyzed for GM in patients fulfilling the host factor criteria as defined by the EORTC/MSG. A cut-off index value GM of ≥0.5 was used. Clinical data, results of chest CT-scans and BAL cultures were collected. Sensitivity, specificity, PPV, and NPV of BAL GM for a diagnosis of proven and probable IPA (n = 41) were 82.4%, 87.5%, 82.4%, and 87.5% respectively. A significant relation was found for BAL GM and abnormal chest CT (P = 0.01). No significant relationship was observed between BAL Aspergillus sp. culture and chest CT (n = 47). BAL GM and serum GM correlated significantly. In 9 out of 12 patients classified as possible IPA, antifungal therapy was continued or started, despite a negative BAL GM. BAL GM test had good diagnostic value in children suspected of IPA. However, the decision to continue or start antifungal therapy was mainly determined by the clinical suspicion of IPA based on chest CT-outcome, serum GM index values and failure of antibiotic therapy.

Systematic Review and Meta-Analysis of Detecting Galactomannan in Bronchoalveolar Lavage Fluid for Diagnosing Invasive Aspergillosis

BackgroundBronchoalveolar lavage (BAL) galactomannan (GM) assay has been used for diagnosing invasive aspergillosis (IA). We aimed to derive a definitive estimate of the overall accuracy of BAL-GM for diagnosing IA.Methods and ResultsWe undertook a systematic review of thirty diagnostic studies that evaluated the BAL-GM assay for diagnosing IA. PubMed and CBM (China Biological Medicine Database) databasees were searched for relevant studies published in all languages up until Feb 2012. The pooled diagnostic odds ratio (DOR) and summary receiver operating characteristic (SROC) were constructed for each cutoff value. Additionally, pooled sensitivity (SEN), specificity (SPE), and positive and negative likelihood ratios (PLR and NLR, respectively) were calculated for summarizing overall test performance. Thirty studies were included in this meta-analysis. The summary estimates of pooled DOR, SEN, SPE, PLR, and NLR of the BAL-GM assay (cutoff value 0.5) for proven or probable IA were 52.7 (95% confidence interval (CI) 31.8–87.3), 0.87 (95% CI 0.79–0.92), 0.89 (95% CI 0.85–0.92), 8.0 (95% CI 5.7–11.1) and 0.15 (95% CI 0.10–0.23) respectively. The SROC was 0.94 (95% CI 0.92–0.96). Compared with cutoff value of 0.5, it has higher DOR, SPE and PLR, and similar SEN and NLR with cutoff value of 1.0, which indicated the optimal cutoff value might be 1.0. Compared with BAL-GM, serum GM has a lower SEN and higher SPE, while PCR displays a lower SEN and a similar SPE.ConclusionWith the optimal cutoff value of 1.0, the BAL-GM assay has higher SEN compared to PCR and serum GM test. It is a useful adjunct in the diagnosis of proven and probable IA.

Comparison of Serum and Bronchoalveolar Lavage Galactomannan in Diagnosing Invasive Aspergillosis in Solid-Organ Transplant Recipients

This study sought to compare the sensitivities of serum galactomannan and bronchoalveolar lavage galactomannan in diagnosing invasive aspergillosis in solid-organ transplant recipients (lung and heart). This study took place in the lung transplant center of the National Research Institute for Tuberculosis and Lung Disease. All patients with clinical and radiologic manifestations suggestive of pulmonary infection were included. Serum and bronchoalveolar lavage galactomannan were measured. Seventeen patients were included (lung, 15; heart, 1; heart-lung, 1). Probable or definite invasive aspergillosis was diagnosed in 9 patients. With a cutoff ≥ 0.5, serum galactomannan sensitivity and specificity for diagnosing invasive aspergillosis were 77.18% and 100%. Negative predictive value and positive predictive value were 80% and 100%. The sensitivity and specificity of bronchoalveolar lavage galactomannan for diagnosing invasive aspergillosis with cutoff of ≥ 0.5 was 100%. Regarding the high levels of mortality and problems in diagnosing this disease, using bronchoalveolar lavage galactomannan could be a suitable option.

Comparison of an Aspergillus Real-time Polymerase Chain Reaction Assay With Galactomannan Testing of Bronchoalvelolar Lavage Fluid for the Diagnosis of Invasive Pulmonary Aspergillosis in Lung Transplant Recipients

Early diagnosis and treatment of invasive pulmonary aspergillosis (IPA) improves outcome. We compared the performance of publicly available pan-Aspergillus, Aspergillus fumigatus-, and Aspergillus terreus-specific real-time polymerase chain reaction (PCR) assays with the Platelia galactomannan (GM) assay in 150 bronchoalveolar lavage (BAL) samples from lung transplant recipients (16 proven/probable IPA, 26 Aspergillus colonization, 11 non-Aspergillus mold colonization, and 97 negative controls). The sensitivity and specificity of pan-Aspergillus PCR (optimal quantification cycle [Cq], ≤35.0 by receiver operating characteristic analysis) and GM (≥.5) for diagnosing IPA were 100% (95% confidence interval, 79%-100%) and 88% (79%-92%), and 93% (68%-100%) and 89% (82%-93%), respectively. The sensitivity and specificity of A. fumigatus-specific PCR were 85% (55%-89%) and 96% (91%-98%), respectively. A. terreus-specific PCR was positive for the 1 patient with IPA due to this species; specificity was 99% (148 of 149 samples). Aspergillus PCR identified 1 patient with IPA not diagnosed by GM. For BAL samples associated with Aspergillus colonization, the specificity of GM (92%) was higher than that of pan-Aspergillus PCR (50%; P = .003). Among negative control samples, the specificity of pan-Aspergillus PCR (97%) was higher than that of BAL GM (88%; P = .03). Positive results for both BAL PCR and GM testing improved the specificity to 97% with minimal detriment to sensitivity (93%). A recently developed pan-Aspergillus PCR assay and GM testing of BAL fluid may facilitate the diagnosis of IPA after lung transplantation. A. fumigatus- and A. terreus-specific real-time PCR assays may be useful in rapidly identifying the most common cause of IPA and a species that is intrinsically resistant to amphotericin B, respectively.

Serum and Bronchoalveolar Lavage Fluid Galactomannan Assays in Patients with Pulmonary Aspergilloma

The sensitivities of the serum and bronchoalveolar lavage galactomannan (GM) assays in 48 patients with pulmonary aspergilloma were 38% (13 of 34; 95% confidence interval [CI], 22%-56%) and 92% (33 of 36; 95% CI, 78%-98%), respectively. The positivity of serum GM assays was significantly higher in patients with hemoptysis than in those without hemoptysis (52% vs 9%; P=.02).

Galactomannan Testing in Bronchoalveolar Lavage Fluid Facilitates the Diagnosis of Invasive Pulmonary Aspergillosis in Patients with Hematologic Malignancies and Stem Cell Transplant Recipients

Invasive pulmonary aspergillosis (IPA) is a major cause of mortality in patients with stem cell transplants and hematologic malignancies. Timely diagnosis of IPA improves survival but is difficult to make. We evaluated the effectiveness of bronchoalveolar lavage (BAL) galactomannan (GM) in diagnosing IPA in these populations by retrospectively reviewing records of 67 consecutive patients, in whom 89 BAL GM tests were performed. For patients with IPA, only the first BAL sample linked to the IPA episode was analyzed. Eighty samples were associated with proven, 12 with probable, and 32 with possible invasive fungal infections (IFI), and 37 were associated with no IFI. Among patients with IFIs, 4 had proven, 11 probable, and 32 possible IPA. Using BAL GM ≥ 0.5 (cutoff for serum GM) and ≥ 0.85 (optimal cutoff identified by receiver-operating characteristic curve), the sensitivity in diagnosing proven or probable IPA was 73% (11/15) and 67% (10/15), respectively, and specificity was 89% (33/37) and 95% (35/37). At these cutoffs, positive and negative predictive values were 73% (11/15) and 83% (10/12), and 89% (33/37) and 87% (35/40), respectively. BAL GM was more sensitive than cytology (0%, 0/14), BAL culture (27%, 4/15), transbronchial biopsy (40%, 2/5), or serum GM (67%, 10/15) for diagnosing IPA. BAL GM was ≥ 0.85 and ≥ 0.5 in 86% (6/7) and 100% (7/7) of patients with proven or probable IPA who received a mold-active agent for ≤ 3 days. BAL GM added sensitivity to serum GM and other means of diagnosing IPA, and was not impacted by short courses of mold-active agents.

Clinical utility and prognostic value of bronchoalveolar lavage galactomannan in patients with hematologic malignancies

We conducted a single-center retrospective cohort study to determine the performance characteristics of the galactomannan (GM) assay in bronchoalveolar lavage (BAL) in patients with hematologic malignancies. Patients were classified as proven, probable, possible, or no invasive pulmonary aspergillosis (IPA), according to international guidelines. A total of 173 BAL samples from 145 patients were included. There were 5 proven, 7 probable, and 35 possible cases of IPA. Using a GM index cutoff of ≥0.5, the sensitivity, specificity, and positive and negative predictive values (PPV and NPV, respectively) of the BAL GM assay were 100%, 78%, 26%, and 100%, respectively. Using a GM index cutoff of ≥2.0, the sensitivity and NPV remained 100%, but specificity and PPV increased to 93% and 50%, respectively. The BAL GM assay is a highly sensitive screening test for IPA in patients with hematologic malignancies. Increasing the cutoff value to 2.0 would improve the performance of this assay.

Aspergillus galactomannan antigen assay in bronchoalveolar lavage fluid for diagnosis of invasive pulmonary aspergillosis

A recently developed bronchoalveolar lavage (BAL) galactomannan (GM) assay shows promising results. We evaluated the diagnostic performance of this assay and analyzed risk factors for false-positive results. A prospective cohort study was performed in a tertiary hospital over a 9-month period. We reviewed all adult patients who underwent GM assays of BAL. Patients were categorized with proven, probable, or possible invasive pulmonary aspergillosis (IPA) according to revised EORTC/MSG definitions. Each patient with a false-positive BAL GM result was matched with three patients with true-negative BAL GM result, and the risk factors for false-positive BAL GM results were determined. Of 359 enrolled patients, 22 (6%) were diagnosed with IPA (1 proven, 17 probable, and 4 possible). Of the 22 patients with IPA, 17 (77%) had already received antifungal agents before the BAL GM assay was conducted. At an index cutoff value of ≥0.5, the BAL GM assay had a sensitivity of 64% (95% CI 41%-83%) and a specificity of 89% (95% CI 85%-92%). However, at an index cutoff value of ≥0.2, the BAL GM assay had a sensitivity of 86% (95% CI 65%-97%) and a specificity of 74% (95% CI 69%-79%). Of the 52 patients with positive BAL GM assay (≥0.5), 25 (7%) were false-positives. Univariate and multivariate analysis revealed that treatment with piperacillin-tazobactam or ampicillin-sulbactam was associated with false-positive BAL GM results. The BAL GM assay appears promising for the diagnosis of IPA. However, treatment with certain antibiotics may interfere with the results of the BAL GM assay.

Diagnosis of Invasive Aspergillosis in Lung Transplant Recipients by Detection of Galactomannan in the Bronchoalveolar Lavage Fluid

Galactomannan (GM) detection in serum samples has been used to diagnose invasive aspergillosis (IA). Limited sensitivity has been observed in lung transplant recipients, for whom bronchoalveolar lavage (BAL) testing has been advocated. Because airway colonization with Aspergillus species occurs frequently in these patients, false-positive GM results have been reported if the cutoff validated for sera is used (i.e., 0.5). Herein, we prospectively studied BAL fluid samples from 60 lung transplant patients to determine the optimal cutoff for BAL GM testing. Only one sample per patient was studied. BAL samples were vortexed and processed according to the manufacturer's instructions for serum samples. Sensitivity, specificity, and likelihood ratios were calculated in reference to proven or probable IA cases using receiver operating characteristic analysis. Eight patients had IA during the study (incidence 13.3%), including four patients with proven IA. Aspergillosis increased 5-fold the risk of death in lung transplant recipients. The positive predictive value of a positive BAL GM test at the 0.5 cutoff was low (24.2%). Raising the cutoff improved test specificity without compromising sensitivity. The best cutoff was defined at 1.5 (sensitivity 100% and specificity 90.4%). This study reinforces the importance of BAL GM testing in lung transplant recipients, particularly to exclude the diagnosis of IA. To minimize the frequency of false-positive results, a higher test cutoff should be applied to BAL samples, in comparison with serum samples.

Galactomannan testing of bronchoalveolar lavage fluid is useful for diagnosis of invasive pulmonary aspergillosis in hematology patients

BackgroundInvasive pulmonary aspergillosis (IPA) is a major cause of morbidity and mortality in patients with hematological malignancies in the setting of profound neutropenia and/or hematopoietic stem cell transplantation. Early diagnosis and therapy has been shown to improve outcomes, but reaching a definitive diagnosis quickly can be problematic. Recently, galactomannan testing of bronchoalveolar lavage (BAL) fluid has been investigated as a diagnostic test for IPA, but widespread experience and consensus on optical density (OD) cut-offs remain lacking.MethodsWe performed a prospective case-control study to determine an optimal BAL galactomannan OD cutoff for IPA in at-risk patients with hematological diagnoses. Cases were subjects with hematological diagnoses who met established definitions for proven or probable IPA. There were two control groups: subjects with hematological diagnoses who did not meet definitions for proven or probable IPA and subjects with non-hematological diagnoses who had no evidence of aspergillosis. Following bronchoscopy and BAL, galactomannan testing was performed using the Platelia Aspergillus seroassay in accordance with the manufacturer's instructions.ResultsThere were 10 cases and 52 controls. Cases had higher BAL fluid galactomannan OD indices (median 4.1, range 1.1-7.7) compared with controls (median 0.3, range 0.1-1.1). ROC analysis demonstrated an optimum OD index cutoff of 1.1, with high specificity (98.1%) and sensitivity (100%) for diagnosing IPA.ConclusionsOur results also support BAL galactomannan testing as a reasonably safe test with higher sensitivity compared to serum galactomannan testing in at-risk patients with hematological diseases. A higher OD cutoff is necessary to avoid over-diagnosis of IPA, and a standardized method of collection should be established before results can be compared between centers.

False-positive serum and bronchoalveolar lavage Aspergillus galactomannan assays caused by different antibiotics

Our objective was to identify false-positive serum and bronchoalveolar lavage (BAL) fluid galactomannan (GM) tests caused by various antibiotics commonly used in general practice. Serum and BAL samples from patients who did not have the diagnostic criteria of invasive aspergillosis and received different antibiotics were prospectively analyzed for GM. Serum and BAL samples were also collected from patients who did not receive antibiotics. At the cut-off index of >or=0.5, false-positive serum results were found in patients who received amoxicillin-clavulanate, piperacillin-tazobactam, cefepime, and cefoperazone-sulbactam (26.7%, 58.3%, 14.3%, and 66.7%, respectively). Fungal colonization in BAL samples had a higher BAL GM than those without fungal colonization. In 71 patients who had a negative BAL culture for fungi, at the cut-off value of >or=1.0, false-positive BAL fluid results were found in patients who received amoxicillin-clavulanate (27.3%), piperacillin-tazobactam (50%), cefepime (16.7%), carbapenem (45.5%), and ceftriaxone (45.5%). False-positive serum and BAL GM assays were also detected in patients who did not receive any antibiotics. In summary, this study demonstrates the false-positive GM levels in serum and BAL caused by beta-lactam antibiotics that are commonly used in general practice. Physicians should be aware of this possible interference.

Bronchoalveolar Lavage Fluid Galactomannan for the Diagnosis of Invasive Pulmonary Aspergillosis in Patients with Hematologic Diseases

Prompt diagnosis of invasive pulmonary aspergillosis (IPA) remains a challenge. Galactomannan (GM) detection in bronchoalveolar lavage (BAL) fluid by the Platelia enzyme immunoassay aims to further improve upon the test's utility by applying it directly to specimens from the target organ. A retrospective analysis of the Platelia assay was performed on BAL samples from 99 evaluable high-risk hematology patients, including 58 with proven or probable IPA. BAL GM demonstrated an improved sensitivity profile (91.3% with an optical density [OD] index cutoff of >or=1.0) in comparison with culture and microscopy (50% and 53.3%, respectively). The diagnostic accuracy as given by the area under the receiver operating characteristic curve was 0.93 (95% confidence interval, 0.88-0.99); further decreasing the OD index cutoff to 0.5 compromised specificity more than it improved sensitivity. Estimates of the positive and negative predictive value of the Platelia assay on BAL samples (OD index, >or=1.0) were 76% and 96%, respectively. The mean BAL GM OD index was not different in neutropenic versus nonneutropenic case patients (3.9 and 4.5, respectively; P = .3); however, a trend toward decreased sensitivity in patients receiving mold-active prophylaxis was noted. BAL GM is a valuable adjunctive diagnostic tool to other conventional microbiologic and radiologic studies.

Combination antifungal treatment of pseudomembranous tracheobronchial invasive aspergillosis: a case report

Invasive aspergillosis (IA) is increasingly recognized in the intensive care unit (ICU), and new risk factors associated with respiratory colonization or infection by Aspergillus spp. include steroid treatment and chronic lung obstructive disease [1, 2]. In a review of 289 autopsies in the ICU, IA was the leading cause of Goldman class I discrepancy (a missed major diagnosis with major impact on patient management and survival) [3]. The epidemiology of IA indicates an increasing number of infections in immunosuppressed patients/individuals undergoing transplantation of bone marrow, hematopoietic stem cells, or organ transplantations, and those receiving intensive chemotherapy or other immunosuppressive treatments. A broad group of patients who are admitted to ICU also have some form of immunosupppression and may be susceptible to invasive mould infections. For various reasons, figures about the true incidence of IA in ICU are difficult to generate. The most important reason is the difficulty encountered in making a definite diagnosis of IA (lack of sensitivity and specificity with regard to culture and radiology) [4]. Recently, galactomannan (GM) in bronchoalveolar lavage (BAL) fluid appears to be a promising tool for early diagnosis in non-neutropenic critically ill patients and has been associated in proven cases with sensitivity and specificity of 88 and 87%, respectively [5]. Pseudomembranous and obstructive Aspergillus tracheobronchitis are still considered to have a fatal outcome and have been reported in a wide variety of patients [6]. There has been only one report in a patient with diabetes which was treated by deoxycholate amphotericin B (AmB) and subsequent addition of oral itraconazole [7]. In this paper, we report a pseudomembranous and obstructive tracheobronchitis in a diabetic patient successfully treated with caspofungin and AmB.

The Role of Bronchoalveolar Lavage Galactomannan in the Diagnosis of Pediatric Invasive Aspergillosis

: Molecular biomarkers such as the galactomannan assay are of increasing interest in clinical settings for diagnosis of invasive aspergillosis (IA). The use of bronchoalveolar lavage galactomannan (BAL GM) is being validated in adult populations, but has not been systematically evaluated in pediatric patients. : A retrospective analysis was performed of patients for whom GM assays from BAL were submitted between November 1, 2006 and November14, 2007 at Childrens Hospital Los Angeles. Medical charts were reviewed and patients were categorized as having no, possible, probable, and proven IA, per established definitions. : Of 85 pediatric patients who had a BAL GM submitted during the study, 59 were immunocompromised. Three patients were identified as having proven IA, 6 had probable IA, 37 had possible IA, and 39 had no evidence of IA; 38 had a concurrent serum GM performed. A positive, linear correlation was established between BAL and serum GM, using OD index values (rho = 0.48, P = 0.002). Among immunocompromised patients, receiver operating characteristic curves demonstrated an optimal BAL GM OD cut-off value of 0.87, that yielded a sensitivity for probable/proven IA of 78% and a specificity of 100%. At 0.87, the positive and negative predictive values among immunocompromised patients were 58% and 96%, respectively. : We found a correlation between BAL GM values and a diagnosis of IA. We also noted a linear relationship between serum and BAL GM values. Receiver operating characteristic curves obtained from our pediatric data validate the current cut off for serum and suggest a possible cut off for BAL specimens.

Performance Characteristics of the Platelia Aspergillus Enzyme Immunoassay for Detection of Aspergillus Galactomannan Antigen in Bronchoalveolar Lavage Fluid

We have evaluated the Platelia Aspergillus enzyme immunoassay for detection of galactomannan in bronchoalveolar lavage (BAL) specimens in solid organ transplant patients with aspergillosis. The precision and reproducibility in serum or BAL to which galactomannan was added were similar. Sensitivity was 81.8% in patients with aspergillosis, and specificity was 95.8% in lung transplant patients who underwent BAL for surveillance for infection or rejection. Among transplant controls, positive results were more common in patients (i) who underwent diagnostic BAL performed for evaluation of symptoms or chest computed tomographic abnormalities, (ii) who had undergone lung transplantation, or (iii) who were colonized with Aspergillus. Galactomannan testing in BAL is useful for diagnosis of aspergillosis in transplant patients. The significance of positive results in patients without confirmed aspergillosis requires further evaluation.

Potential voriconazole and caspofungin sequestration during extracorporeal membrane oxygenation

Sir: A 31-year-old woman (weighing 50 kg) was admitted to our ICU for cardiogenic shock caused by fulminant myocarditis. Twelve hours later, extracorporeal oxygenation membrane (ECMO) support via percutaneous arterial and venous femoral cannulae was initiated; the system comprised a membrane oxygenator (Quadrox Bioline, Jostra-Maquet, Orleans, France) and a centrifugal pump (Rotaflow, Jostra-Maquet). Continuous venovenous hemodialfiltration (PRISMA machine; respective blood, dialysate and ultrafiltration flows: 120 ml/min, 500 ml/h and 1,000 ml/h) was started because of acute renal failure. Six days later, Aspergillus fumigatus was isolated from pulmonary secretions, obtained via bronchoalveolar lavage (BAL), and extensive tracheal pseudomembranous lesions. Although her serum was galactomannan-antigen-negative (enzyme immunoassay, Bio-Rad), the BAL galactomannan index was 4.8 (threshold, 0.5). Combined intravenous (IV) antifungal therapy was initiated with voriconazole [6 mg/kg bid day 1, then 4 mg/kg bid] and caspofungin (70 mg day 1, then 50 mg/day). Because of persistent A. fumigatus-positive tracheal lesion cultures on D11, respective antifungal doses were increased to 8 mg/kg bid and 70 mg/day. On D14, persistent extensive tracheal pseudomembranous lesions, positive BAL and tracheal aspirate cultures and a BAL galactomannan index of 8 indicated treatment failure. On D15, voriconazole was stopped, and based on in vitro results [1], caspofungin was combined with IV liposomal amphotericin B (3 mg/kg/ day) and flucytosine (1.5 g/day); nebulized liposomal amphotericin B (25 mg bid) was also administered. The patient was weaned from ECMO 21 days after ICU admission. BAL cultures became negative 16 days after starting the three-drug regimen, followed shortly thereafter by an undetectable galactomannan index. The patient was discharged from the ICU on D53. During ECMO, voriconazole, caspofungin and amphotericin B levels in blood collected from the indwelling arterial line were determined by high-performance liquid chromatography (respective quantification limits: 0.2, 0.5 and 0.1 mg/l; see Table 1). ECMO improves outcomes of patients with fulminant myocarditis [2]. During ECMO, drugs exhibit complex pharmacokinetics due to the larger volume of distribution, and their binding to various extracorporeal circuit components may markedly alter the pharmacokinetics of commonly used agents [3]. Molecular size, degree of ionization and physicochemical drug properties, e.g., the octanol–water partition coefficient, which is associated with a drug’s lipophilicity, may influence the degree of binding to plastics [3]. A recent ex vivo study [4] demonstrated a 71% ‘loss’ of voriconazole, a finding consistent with our very low or undetectable circulating levels, despite high doses, that is clinically relevant because of the significant association between trough blood voriconazole levels and outcome [5]. A renal replacement role was unlikely because nonhigh-volume hemodialfiltration contributes minimally to voriconazole elimination. ECMO also affected caspofungin pharmacokinetics, while blood amphotericin B levels remained within the therapeutic range. To the best of our knowledge, no data are available concerning blood caspofungin concentrations in patients on ECMO. In such patients, because adequate blood voriconazole and caspofungin concentrations may not be Table 1 Blood levels of antifungals during ECMO

A Novel Non-Invasive Method of Detecting Galactomannan in Suspected Pulmonary Aspergillosis.

Invasive pulmonary aspergillosis (IPA) is now the leading cause of infectious mortality in stem cell transplant recipients. Despite recent advances in molecular diagnostic techniques the diagnosis remains problematic. The mycological yield from bronchoalveolar lavage (BAL) is poor, especially if the patient has been exposed to antifungal drugs. Galactomannan (GM), a component of Aspergillus' cell wall is detectable in serum and BAL fluid in IPA, though its role in the latter has not been validated. Exhaled breath condensate (EBC) collection provides a non-invasive means of sampling the airway lining fluid. This method has been used in respiratory disease to detect markers of inflammation and oxidative stress. EBC is collected by breathing into a tube surrounded by a cooled insulated metal sleeve (RTube®, Respiratory Research, USA) and results in 1–2 millilitres of fluid. The application of EBC detection of volatile and non-volatile organic compounds has not been explored in this setting. We report on pilot data of the detection of GM in EBC from a subset of patients at high risk of infection forming part of a prospective study into the early diagnosis of IPA in allogeneic transplant recipients and patients with acute leukaemia. Patients were recruited prior to commencing treatment for their underlying haematological condition; the study period finished at neutrophil recovery (> 0.5 x 109/L) or discharge from hospital. EBC and serum were collected once and twice weekly respectively, throughout the study period. High resolution CT (HRCT) imaging of the chest was carried out after 72 hours of neutropenic fever unresponsive to broad-spectrum antibiotics. Where possible, BAL was carried out in the most affected lobe in patients with an abnormal HRCT chest. GM was measured in EBC, serum and, where applicable, BAL fluid using a commercially available kit (Platelia® Aspergillus, Bio-Rad, France). The likelihood of IPA was assigned using criteria published by European Organisation for Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG) by 2 investigators blinded to clinical details (Ascioglu et al, CID 2002). Of 39 patients, 6 had proven/probable IPA, 9 possible and 24 had no evidence of IPA. Four of the 6 patients with proven/probable disease had GM-positive BAL fluid; 3 of these 4 had persistently negative GM in serum. In 1 of the 6, the EBC GM was positive 3 weeks earlier than the development of HRCT evidence of IPA and a positive BAL GM. In another, the EBC GM was positive early in the infection, whilst BAL GM remained negative. EBC and serum GM followed the same trend in all 6 patients, although the values for both EBC and serum remained below the currently recognised positive cut-off of 0.5 in 2 patients. In patients with possible or no evidence of IPA, GM was negative in all but 2 of 111 EBC samples. These results suggest that GM can be detected in EBC and follows the same trend as in serum. It may even predate GM detection in BAL fluid or abnormal HRCT signs, thereby obviating the need for more invasive investigations. Elevated EBC GM levels early in the course of neutropenia may represent infection and could be applied to the employment of pre-emptive treatment strategies.

Detection of Aspergillus fumigatus‐specific DNA, (1–3)‐β‐d‐glucan and galactomannan in serum and bronchoalveolar lavage specimens of experimentally infected rats

The aim of this study was to detect Aspergillus fumigatus-specific DNA by nested PCR (nPCR) in serum and bronchoalveolar lavage (BAL) specimens of experimentally infected rats and compare the results with (1-3)-beta-D-glucan (BDG) and galactomannan (GM) detection. Sixty Wistar rats, immunosuppressed with an intraperitoneal injection of cyclophosphamide (70 mg kg(-1)) were infected with 1 x 10(6)A. fumigatus conidia. The rats were killed on days 1, 3, 5, 7 and 9 postinfection in groups of six each and their BAL, blood and lungs were cultured. The A. fumigatus-specific DNA, BDG and GM in serum and BAL were detected by nPCR, Fungitell kit and Aspergillus Platelia kit respectively. Base line values were obtained by using sera from six healthy rats. Except the lungs, blood and BAL specimens of all the infected rats were negative for A. fumigatus culture. The BDG, GM and nPCR positivity in serum specimens was 80%, 77% and 63% respectively. The sensitivity of GM and nPCR tests in BAL specimens was 77% and 70% respectively. The data suggest that BDG and GM appear early in the course of infection, and have similar kinetics (r = 0.483, P = 0.007). Hence, their combined detection could be useful in the early diagnosis of invasive aspergillosis.

Galactomannan in Bronchoalveolar Lavage Fluid

Invasive aspergillosis (IA) is an important cause of mortality in patients with hematologic malignancies. However, IA appears to be gaining a foothold in the intensive care unit (ICU) in patients without classical risk factors. A recent study described 89 cases of IA in patients in a medical ICU without leukemia or cancer. The diagnosis of IA remains difficult and is often established too late. Galactomannan (GM) is an exo-antigen released from Aspergillus hyphae while they invade host tissue. This prospective single-center study was conducted to investigate the role of GM in bronchoalveolar lavage (BAL) fluid as a tool for early diagnosis of IA in the ICU. All patients with risk factors identified in our earlier study were evaluated. BAL for culture and GM detection, serum GM levels, and computed tomography scan were obtained for all included patients with signs of pneumonia. Patients were classified as having proven, probable, or possible IA. A total of 110 patients out of 1,109 admissions were eligible. There were 26 proven IA cases. Using a cutoff index of 0.5, the sensitivity and specificity of GM detection in BAL fluid was 88 and 87%, respectively. The sensitivity of serum GM was only 42%. In 11 of 26 proven cases, BAL culture and serum GM remained negative, whereas GM in BAL was positive. IA is common in immunocompromised, critically ill patients. GM detection in BAL fluid seems to be useful in establishing or excluding the diagnosis of IA in the ICU.