Fungal nucleic acid detection for invasive aspergillosis

A universal PCR-assay for the detection of fungal DNA was compared with microscopy and culture for the diagnosis of invasive aspergillosis using 78 samples from 42 patients. Eighteen patients were suffering from invasive aspergillosis, 5 patients were colonized with Aspergillus in the respiratory tract, 19 patients did not show any sign of aspergillosis. Samples from 6 of the 18 patients with invasive aspergillosis were microscopically positive with true mycelia, 15 of 18 grew Aspergillus in culture, 16 of 18 were PCR-positive. The combination of microscopy and culture led to the diagnosis in 17 of 18 patients, the combination of microscopy and PCR in 16 of 18 and the combination of culture and PCR in all the 18 patients. For 3 of 18 patients, PCR was the diagnostic key: in 2 biopsies the histologically detected fungal elements were identified as Aspergillus, in 3 bronchial lavages from 1 patients nothing but PCR was positive for Aspergillus. Four out of 5 culture positive patients with Aspergillus colonization were also PCR positive; one out of 19 patients without aspergillosis was culture positive, 3 out of 19 were falsely PCR positive. Candida colonization in the upper respiratory tract or Pneumocystis carinii pneumonia did not lead to false positive Aspergillus-PCR results. In conclusion, the evaluated fungal PCR-assay can supplement conventional methods for the diagnosis of invasive aspergillosis.     

The Platelia Aspergillus ELISA in diagnosis of invasive pulmonary aspergilosis (IPA).

The sensitivity of a sandwich enzyme-linked immunosorbent assay (ELISA) for detecting Aspergillus galactomannan was evaluated with 66 serum samples and 113 specimens of the respiratory tract obtained from 52 patients with pulmonary diseases. The patients were divided into five groups: proven invasive pulmonary aspergillosis (IPA) (five patients), probable IPA (seven patients), Aspergillus colonization (eight patients) or unlikely Aspergillus infection (27 patients). Another five patients with doubtful diagnostic test results are discussed in detail. The results of the Platelia Aspergillus ELISA (Sanofi Pasteur, Freiburg, Germany) in testing specimens of the respiratory tract were 90% sensitivity in proven (serum 38%), 60% in probable (serum 37%) and 71% in Aspergillus colonization (serum 0%). Furthermore, 85% of the Aspergillus spp. from positive cultures of specimens of the respiratory tract were also detected in the ELISA. A total of 57% of the culture negative specimens of patients with a least one positive culture or proven aspergillosis in a series of specimens were positive in the ELISA.     

Therapy of invasive aspergillosis with itraconazole: improvement of therapeutic efficacy by early diagnosis

We report on the treatment of invasive aspergillosis with the new triazole antimycotic agent itraconazole. All 11 patients suffered from pulmonary invasive aspergillosis. Two patients also had cerebral aspergillosis; in one of these patients the paranasal sinuses were also invaded. Underlying diseases were acute lymphoblastic leukaemia (n = 3), acute myeloid leukaemia (n = 4); one patient underwent allogeneic bone marrow transplantation before he developed aspergillosis; another was transplanted after successful aspergillosis treatment, liver cirrhosis (n = 1), lung infarction after pulmonary embolism (n = 1), chronic bronchitis after pulmonary tuberculosis (n = 1) and AIDS (n = 1). In five cases initial diagnosis was established by means of mycological methods and clinical signs. In six patients invasive pulmonary aspergillosis was initially diagnosed due to the clinical criteria presented in this paper. Secondary mycological confirmation after onset of therapy was achieved in five out of these six patients. All of the patients initially responded to therapy. One female patient experienced a relapse of aspergillosis and died of cerebral involvement and relapsing leukaemia. Two further patients died due to underlying diseases (pulmonary embolism, relapsing leukaemia). Nine patients (82%) were cured of the mycosis, including the patient with cerebral involvement; two underwent surgical resection of residual pulmonary lesions. Itraconazole is a very effective drug for treatment of invasive aspergillosis. Therapeutic efficacy can be optimized by early diagnosis using clinical criteria and prompt start of treatment.     

Invasive aspergillosis: results of an 8-year study

We analysed retrospectively 90 cases of invasive aspergillosis (IA) which occurred at the University Hospital and the Thoraxklinik gGmbH Heidelberg between 1991 and 1998. 71 cases were histologically proven, 19 were probable diseases. There were 49 male and 41 female patients, with a mean age of 51.5 years (range 16 days to 80 years). Underlying diseases were: hematological malignancies in 52% (n = 47; 24 with acute leukemia), solid organ transplantation (n = 11; 9 liver, 1 kidney, 1 heart), solid cancer (n = 10), others (n = 21), and in one case no underlying disease was diagnosed. Only 54 cases (60%) were correctly diagnosed as IA during lifetime of the patients. In 59 cases (65%) only the lung was affected, 25 patients suffered from disseminated IA, in 6 patients only extrapulmonary lesions were present. 11 patients underwent lung surgery, 63 patients received antimycotic drugs (44 amphotericin B, 15 fluconazole, 4 itraconazole), 21 were not treated antimycotically. 68 patients (71%) died, from these 30 (36%) due to IA during remission of the underlying disease. The laboratory methods showed the following sensitivities, respectively: microscopy by calcofluor white staining 17%, culture 69%, Aspergillus-PCR from respiratory tract samples and biopsies 95%, galactomannan antigen detection by latex agglutination 28%, by enzyme immunoassay 59%, Aspergillus antibody detection 23%.     

Surveillance of invasive mold infections in lung transplant recipients: effect of antimycotic prophylaxis with itraconazole and voriconazole

Between January 2002 and December 2003 all 157 patients (pts) that underwent lung transplantation (LTx) at our institution were prospectively screened for invasive aspergillosis (IA) during their perioperative hospital stay. Patients were regarded as IA positive, if they met the EORTC criteria for 'probable' or 'proven' IA. Records of pts were screened retrospectively for antimycotic prophylaxis. Eight of the 157 pts developed 'probable' or 'proven' IA (5.1%) within 17 +/- 10 days after LTx. This was associated with a 14-fold increased mortality compared with all pts without aspergillosis (P < 0.01, OR 13.8, CI(95%) 2.5-82). Preoperative colonization with Aspergillus was a significant risk factor for IA (P < 0.001, OR 21.9, CI(95%) 4.9-97). We switched our prophylactic strategies to the primary administration of voriconazole in high risk pts (pre-LTx colonization) starting in December 2002. Six pts (6%) of 101 pts receiving itraconazole for antimycotic prophylaxis beginning at postoperative day (POD) one developed IA, of which three pts showed cerebral aspergillosis. One pt (5%) of 18 pts receiving voriconazole prophylaxis developed IA, while 10 pts showed pretransplant colonization with Aspergillus species. Thirty-eight pts received itraconazole prophylaxis at a later time point (>POD 14). By switching our prophylactic strategy to the use of voriconazole in high risk pts, we have decreased the incidence of IA from 8% (six of 75) in 2002 to 2% (two of 82) in 2003. This study shows a high incidence of IA during the very early postoperative course after LTx of 5%. This is associated with a significantly increased risk for mortality. Voriconazole prophylaxis appears to be superior to itraconazole, especially in high risk pts with pretransplant Aspergillus colonization.     

Clinical characteristics of 45 patients with invasive pulmonary aspergillosis

BACKGROUND:

Invasive aspergillosis (IA) is a common complication in patients with hematologic malignancies. Patients with solid tumors also are at risk for IA because they may develop neutropenia as a result of chemotherapy and radiotherapy. However, studies of IA in patients with solid tumors are rare. In this study, the risk factors and clinical characteristics of pulmonary infection and death mediated by invasive pulmonary aspergillosis (IPA) as complications in patients with lung cancer were determined.

METHODS:

The authors conducted a retrospective analysis of the clinical notes from 45 patients who had IPA.

RESULTS:

Among 1711 patients with lung cancer, 45 patients contracted pulmonary aspergillosis (2.63%). There were 10 cases of proven disease and 35 cases of probable disease. In univariate analysis, the main predisposing factors were clinical stage IV disease (P = .018), chemotherapy during the month preceding infection (P = .033), and corticosteroid use (≥3 days; P = .038). In multivariate analysis, only clinical stage IV disease (P = .018) was associated with IPA. Furthermore, the mortality rate among lung cancer patients who had pulmonary aspergillosis was 51.1% (23 of 45 patients). Of the patients who died, corticosteroid therapy (P = .001) and grade 3/4 neutropenia (P = .013) were correlated statistically with pulmonary aspergillosis in patients with lung cancer.

CONCLUSIONS:

In univariate analysis, the risk factors for IPA in lung cancer included chemotherapy and corticosteroid use in the month preceding infection and clinical stage IV disease. However, in multivariate analysis, only clinical stage IV disease was identified as a risk factor for IPA. Cancer 2009. © 2009 American Cancer Society.     

Non-value of Aspergillus antigen detection in bronchoalveolar lavage fluids of patients undergoing bone marrow transplantation

Summary. A commercially available antigen assay (Pastorex Aspergillus ) was used to detect Aspergillus antigen in serial bronchoalveolar lavage (BAL) fluids and sera of patients undergoing bone marrow transplantation (six patients with autopsy-proven aspcrgillosis and 10 control patients without evidence of fungal infection). Aspergillus antigen was not detected in 17 BAL fluids of the six patients with proven aspergillosis. In two of the six patients the assay gave positive results in serum specimens. Three of the 10 control patients showed reactive BAL fluids. It is concluded that the latex agglutination assay of BAL fluids has no value in the diagnosis of invasive (pulmonary) aspergillosis in patients undergoing bone marrow transplantation.
Zusammenfassung. Ein kommerziell erhältlicher Test zum Nachweis von Aspergillus -Antigen (Pastorex Aspergillus ) wurde verwendet, um Aspergillus -Antigen in bronchoalveolären Lavage (BAL)-Flüssigkeiten und Seren von Patienten nach Knochenmarktransplantation (sechs Patienten mit autoptisch gesicherter Aspergillose und zehn Kontroll-Patienten ohne Hinweise auf eine Pilz-Infektion) nachzuweisen. Aspergillus -Antigen wurde in 17 BAL-Flüssigkeiten von sechs Patienten mit histologisch gesicherter Aspergillose nicht nachge-wiesen. Bei zwei der sechs Patienten ergab die Untersuchung von Seren positive Resultate. BAL-Flüssigkeiten von drei der zehn Kontroll-Patienten zeigten ein positives Resultat. Der Nachweis von Aspergillus -Antigen in BAL-Flüssigkeiten hat somit keine diagnostische Relevanz für eine invasive (pulmonale) Aspergillose bei Patienten nach Knochenmarktransplantation.     

Aspergillus detection in bronchoscopically acquired material. Significance and interpretation

Pulmonary invasive aspergillosis is frequently difficult to diagnose. In particular, the value of the cultivation of Aspergillus and the Aspergillus galactomannan antigen detection (Pastorex) in bronchoscopically acquired material (bronchoalveolar lavage = BAL, bronchial lavage = BL) in the course of diagnosing this mycosis is viewed controversially. Between January 1996 and September 1999, we obtained 114 positive results in 100 bronchoscopically aquired specimens from a total of 69 patients. 59 of the 69 patients were immunosuppressed, 42 suffered from pulmonary aspergillosis and 38 suffered from invasive pulmonary aspergillosis. The positive prediction rate for a positive result with regard to pulmonary aspergillosis in bronchoscopically acquired material was approximately 61%. Cultivation of Aspergillus was more successful in BAL, and the Aspergillus antigen detection was more successful in BL.     

Case reports. Invasive pulmonary aspergillosis in non-neutropenic patients treated with liposomal amphotericin B

We report two cases of invasive pulmonary aspergillosis due to Aspergillus flavus in one patient who with chronic nephritis and to A. fumigatus in another with malignant lymphoma. After receiving intravenous liposomal amphotericin B therapy for 31 and 35 days, respectively, the patients were cured and did not experience any severe adverse effects.     

Endomyocardial and pericardial aspergillosis in critically ill patients

Invasive aspergillosis(IA) is a potentially lethal complication of Aspergillus infection affecting mainly immunocompromised hosts; however, during the last two decades its incidence was increasingly observed in critically ill immunocompetent patients. The objective of this study is to describe the clinical characteristics of histologically proven endomyocardial and pericardial invasion, in the context of IA, in critically ill patients. Eight critically ill patients with histopathological confirmation of endomyocardial/pericardial aspergillosis were evaluated. Risk factors, clinical and laboratory characteristics, treatment, histopathological characteristics and mortality were recorded. Signs and symptoms of cardiac dysfunction were not observed in any of the patients. Therapy was administered to six of them shortly after the first positive culture. The observed histopathological lesions included haemorrhagic lesions, small vessels with central thrombosis and surrounding consolidated tissue with necrosis. Voriconazole, caspofungin, lipid amphotericin B and itraconazole were the used antifungals. The mortality rate was high (87.5%). Endomyocardial and pericardial aspergillosis are devastating complications of invasive aspergillosis. Clinical suspicion is low making the diagnosis difficult, therefore histopathological examination of tissues are required. The mortality is high.     

An evaluation of nebulised amphotericin B deoxycholate (Fungizone®) for treatment of pulmonary aspergillosis in the UK National Aspergillosis Centre

Antifungal treatment options for allergic bronchopulmonary aspergillosis (ABPA) and severe asthma with fungal sensitisation (SAFS) are largely limited to itraconazole based on the outcome of randomised controlled trials. It is unclear if nebulised amphotericin B deoxycholate (Fungizone^®) is a viable therapeutic option. We evaluated the safety and efficacy of nebulised Fungizone^® in the long‐term treatment of various forms of pulmonary aspergillosis. We assessed the records of 177 patients with various forms of pulmonary aspergillosis attending the National Aspergillosis Centre in Manchester who had received Fungizone^®. Patients first received a challenge test with nebulised Fungizone^® in hospital with spirometry pre/post‐Fungizone^® and nebulised salbutamol given pre‐Fungizone^®. Tolerability and changes in Aspergillus IgE, Aspergillus IgG and total IgE were evaluated. Sixty‐six per cent (117/177) were able to tolerate the test dose of Fungizone^® and in all cases, the reason for discontinuation of the first test dose was worsening breathlessness. Twenty six (21%) stopped therapy within 4‐6 weeks, and the commonest reason cited for discontinuation of therapy was increased breathlessness, hoarseness and cough. Eighteen (10.2%) patients continued the Fungizone^® for >3 months of which 5 (27.8%) recorded an improvement in total IgE, Aspergillus‐specific IgE and Aspergillus IgG. Eleven had ABPA, four had SAFS, two had Aspergillus bronchitis and one had Aspergillus sensitisation with cavitating nodules. Among these 18 patients, sputum fungal culture results went from positive to negative in five patients, became positive in one patient, remained positive in three patients, and remained negative in seven patients. Nebulised Fungizone^® appears to be a poorly tolerated treatment for pulmonary Aspergillosis with high dropout rates. There appears to be both clinical and serological benefits following sustained treatment with nebulised Fungizone^® in some patients.     

Favorable outcome of invasive aspergillosis in patients with acute leukemia

During a 2-year period, 15 of 110 patients (14%) admitted for intensive therapy of acute leukemia associated with prolonged deep granulocytopenia developed documented invasive aspergillosis (IA). Antemortem diagnosis was accomplished in 14, and 13 of 15 (87%) survived the infection. Because of the high success rate, we reviewed the courses of the 15 patients to assess factors associated with this favorable outcome. Eleven presented with pulmonary IA; early symptoms occurred at a mean 21.6 days of granulocytopenia (less than 100/muL) and included refractory fever in 14 and pulmonary signs or symptoms in 11. Primary necrotic chest wall lesions associated with Hickman catheters developed in four at a mean 11 days of granulocytopenia, followed by pulmonary involvement. All 15 patients had chest radiographs during granulocytopenia, with 14 (93%) demonstrating pulmonary infiltrates and/or nodules at a mean 20.6 days of aplasia. Nine patients had lung computerized tomography (CT) scans, revealing nodular infiltrates in one patient and a characteristic zone of low attenuation surrounding a mass-like infiltrate in seven other patients, which was found to be diagnostic of IA. Subsequent CT scans performed during and following bone marrow recovery showed progression to cavitation followed by either complete resolution or minimal pulmonary scarring. Eleven patients developed IA during empiric amphotericin B (Amp-B) therapy (0.5 mg/kg/d) for fever refractory to antibacterial antibiotics. Fourteen patients received high-dose Amp-B (1.0 to 1.5 mg/kg/d), which was started within a mean of 2.2 days of first clinical findings; 13 survived. Ten patients received 5-fluorocytosine in addition to high dose amp-B. Survival was similar regardless of presentation, as 91% with primary pulmonary IA and 75% presenting with chest wall lesions survived. All 13 surviving patients had complete granulocyte recovery at a mean 33.8 days. Nephrotoxicity (creatinine greater than 2.0 mg/dL) was observed in seven patients during therapy for IA, but was transient in all seven. We conclude IA can be successfully treated in the deeply granulocytopenic patient provided that it is recognized and treated early, and provided that antifungal therapy is aggressive and is continued until granulocyte recovery occurs.     

Aspergillus galactomannan detection in the diagnosis of invasive aspergillosis in cancer patients.

PURPOSE: To assess the Aspergillus galactomannan enzyme-linked immunosorbent assay (ELISA) in the diagnosis of invasive aspergillosis (IA) in adult and pediatric oncohematologic patients. PATIENTS AND METHODS: The study was conducted in four patient groups: those with fever of unknown origin (FUO) during neutropenia, suspected pulmonary infection (PI), or nonpulmonary aspergillosis (NPA) and those undergoing surveillance (S) after hematopoietic stem-cell transplantation (HSCT). IA was classified as definite, probable, or possible, according to European Organization for Research and Treatment of Cancer/Mycosis Study Group definitions. RESULTS: A total of 3,294 serum samples were collected during 797 episodes (FUO, 261; PI, 297; NPA, 28; and surveillance, 211), and 153 episodes of IA were diagnosed (31 definite, 67 probable, and 55 possible). Three episodes were first suspected from galactomannan ELISA; the remaining 150 cases were diagnosed from clinical or radiologic evidence. Sensitivity of the ELISA was 64.5%, 16.4%, and 25.5% in definite, probable, and possible episodes of IA, respectively, and was lower in patients positive for anti-Aspergillus antibodies than in antibody-negative patients. Most false-positive results occurred in children and in allogeneic HSCT (allo-HSCT) patients. Overall specificity of the ELISA was 94.8%. It was lower in children compared with adults (P <.0001) and in allo-HSCT patients compared with non-allo-HSCT adults (P =.0002). Lowering the ELISA cutoff value from 1.500 to 0.700 seemed more relevant for non-allo-HSCT adults (sensitivity, 73.1%, 44.3%, and 44.7% in definite, probable, and possible IA, respectively; specificity, 94%). CONCLUSION: Galactomannan ELISA seems less sensitive than previously described, and sensitivity can be further reduced by the presence of anti-Aspergillus antibodies. A new cutoff value for the ELISA of 0.700 is proposed for non-allo-HSCT adults.     

Opportunistic Invasive Fungal Infections Mimicking Progression of Non–Small-Cell Lung Cancer

BackgroundMany studies have shown that invasive pulmonary aspergillosis, cryptococcosis, and mucormycosis can mimic radiographic and clinical features of primary lung cancer. However, more research surveying the incidence and outcomes of these fungal infections among patients with a history of lung cancer is needed. The aim of this study was to describe the occurrence and clinical outcomes of opportunistic invasive fungal infections that can mimic tumors in non–small-cell lung cancer patients.Patients and MethodsPatients seen at Stanford University Medical Center from January 1, 2007, to May 1, 2020, with pulmonary aspergillosis, cryptococcosis, or mucormycosis after non–small-cell lung cancer (NSCLC) diagnosis were reviewed. The European Organization for Research and Treatment of Cancer National Institute of Allergy and Infectious Diseases Mycoses Study Group criteria was used to classify patients with evidence of proven or probable invasive fungal infection within our cohort.ResultsA total of 12 patients with proven or probable invasive mold infection (including 8 cases of aspergillosis) and 1 patient with proven cryptococcosis were identified, without any cases of mucormycosis. Of this cohort, 6 patients (46%) showed radiographic findings that were found to be most consistent with lung cancer by radiologists. Eight cases (62%) were suspected of cancer recurrence or progression by the treatment team on the basis of additional considerations of medical history and clinical symptoms. Most patients had active NSCLC or had a history of recurrence without active NSCLC at the time of fungal discovery (11 patients; 85%). Most patients died without full recovery (7 patients; 54%).ConclusionsInvasive pulmonary aspergillosis and cryptococcosis can often be mistaken as cancer recurrence or progression in patients with a history of NSCLC because of mimicking radiographic and clinical characteristics.     

Potential of polymerase chain reaction and galactomannan for the diagnosis of invasive aspergillosis in patients with febrile neutropenia

The incidence of invasive aspergillosis (IA) has increased over the last years, especially in immuncompromised patients with high mortality rates. Because of difficulties about the diagnosis; serological methods [galactomannan (GM) antigen test] and polymerase chain reaction (PCR) developed in recent years. MycAssay Aspergillus PCR performance in the diagnosis of IA was evaluated and compared with the GM and in‐house PCR. This study was conducted with 358 serum samples obtained from 99 patient with febrile neutropenic episodes who were followed in haematology and bone marrow transplantation units. Patients were classified by the European Organization for the Research and Treatment of Cancer/Mycoses Study Group criteria, 18 of them is proven and probable IA. GM antigen test and two different real‐time PCR; one of them is fist commercial PCR for IA; Mycassay Aspergillus and the other one is in‐house real‐time PCR performed. Sensitivity values were Mycassay Aspergillus PCR, in‐house PCR, and GM 65.38%, 11.53% and 23.07%, respectively. The high sensitivity obtained from Mycassay Aspergillus PCR and sensitivity is increased by using a combination of diagnostic methods. GM antigen test and real‐time PCR could be beneficial for early diagnosis and treatment of IA. For routine usage of PCR as diagnostic assay more studies needed in future.     

Invasive pulmonary aspergillosis in neutropenic patients and the influence of hospital renovation

To evaluate the effects of airborne Aspergillus contamination during and after the renovation work of a Florentine haematology unit, we conducted (November 2003-January 2005) a strict programme of environmental fungal surveillance. Air samples were taken from patients' rooms, along the corridors inside the wards, along the corridor between wards and outside the building. The concentration of Aspergillus fumigatus was high along the corridor between the two haematology wards (2.98 CFU m(-3)), lower in the non-neutropenic patients' rooms and outside the hospital building (1.53 and 1.42 CFU m(-3), respectively), very low in the neutropenic patients' rooms (0.09 CFU m(-3)). During this period, three proven cases (A. fumigatus), two probable ones and two possible cases of invasive pulmonary aspergillosis were documented in 97 patients with acute leukaemia (7%). The three cases of proven aspergillosis coincided with the period of renovation work and with the period in which we have found the maximum concentration of A. fumigatus along the corridor. These data suggest a possible relationship between environmental fungal contamination and the incidence of invasive aspergillosis, and underline the importance of environmental surveillance.     

Invasive aspergillosis: epidemiology and environmental study in haematology patients (Sfax,Tunisia)

Invasive aspergillosis (IA) is a major opportunistic infection in haematology patients. Preventive measures are important to control IA because diagnosis is difficult and the outcome of treatment is poor. We prospectively examined the environmental contamination by Aspergillus and other fungal species and evaluated the prevalence of invasive aspergillosis in the protect unit of haematology. A three‐year prospective study (December 2004–September 2007) was carried out in the department of haematology of Hedi Chaker Hospital. Suspected invasive aspergillosis cases were reviewed and classified as proven, probable and possible invasive aspergillosis using the EORTC criteria. During the study period, we collected weekly environmental samples (patient’s rooms, tables and acclimatisers) and clinical samples from each patient (nasal, expectoration and auricular). Among 105 neutropenic patients, 16 had probable and 13 had possible IA. A total of 1680 clinical samples were collected and A. flavus was most frequently isolated (79.2%). Analysis of 690 environmental samples revealed that Penicillium (44%) was the most frequent followed by Cladosporium (20%), Aspergillus spp. (18%) and Alternaria (13%). The PCR‐sequencing of 30 A. flavus isolates detected from clinical and environmental samples confirmed the mycological identification. Our findings underline the importance of environmental surveillance and strict application of preventive measures.     

Polymerase chain reaction on blood for the diagnosis of invasive pulmonary aspergillosis in cancer patients

BACKGROUND: The premortem diagnosis of invasive pulmonary aspergillosis (IPA) is difficult to make and often missed. Several retrospective studies have suggested that Aspergillus polymerase chain reaction (PCR) performed on serum or whole blood is useful in diagnosing IPA. Two prospective studies confirmed this finding but included a small number of IPA cases. METHODS: The current study was performed to determine the diagnostic role of Aspergillus PCR performed on whole blood specimens from 54 patients with cancer and pulmonary infiltrates for which bronchoscopy with fungal stains and cultures were performed. PCR through amplified Aspergillus mitochondrial DNA and alkaline protease genes was performed on whole blood samples. RESULTS: The cohort in the current study was comprised of 4 patients with definite IPA, 7 patients with probable IPA, 7 patients with possible IPA, and 36 control patients with no evidence of IPA. The sensitivity, specificity, and positive and negative predictive values all were 100% for definite IPA cases and 57%, 100%, 100%, and 92%, respectively, for each of the probable and possible IPA cases. CONCLUSIONS: Aspergillus PCR on whole blood samples is highly sensitive for the detection of IPA and is predictive for IPA. The sensitivity appears to be correlated with the certainty of diagnosis as proven by tissue invasion.     

Aspergillus pericarditis: clinical and pathologic features in the immunocompromised patient

There has been a proliferation of infectious complications due to Aspergillus in patients receiving chemotherapy for cancer and transplantation; however, aspergillus pericarditis has been rarely described. Reported here are the clinical and pathologic findings of Aspergillus pericarditis in six immunocomprised patients who came to autopsy in the past 11 years. Five had leukemia, one had received a renal transplant. All had pulmonary aspergillosis. Two had clinically overt pericarditis leading to cardiac tamponade and death. Pulmonary aspergillosis preceded development of pericardial signs. Chest radiographs, serial electrocardiograms, and echocardiograms showed abnormality but were nonspecific. Pericardiocentesis was performed in one patient but proved nondiagnostic and yielded only transient hemodynamic improvement; postmortem Gram stain of the spun sediment of that pericardial fluid revealed branched hyphae. Although five patients received Amphotericin B, whether it entered the pericardial space is uncertain. Postmortem examination revealed extensive pericardial involvement by Aspergillus associated with effusions as large as 1000 ml Aspergillus penetrated the pericardium by rupture of myocardial abscesses and invasion from contiguous pulmonary foci into the pericardial space. A clinical diagnosis of Aspergillus pericarditis was never established, and at least two died of their pericardial disease. Aspergillus pericarditis is a lethal cardiac infection, which is likely to increase in frequency, and should be considered in the hemodynamically unstable immunocompromised patient, especially when signs of pericarditis or pulmonary aspergillosis are present.     

Intracellular oxidative response of human monocytes and granulocytes to different strains of Aspergillus fumigatus

Aspergillus fumigatus is one of the most prevalent airborne fungal pathogens, causing severe and often fatal infections. Its fungal virulence factors have not been clearly identified. Reactive oxygen species produced by phagocytic cells are potent fungicides for A. fumigatus. The aim of this study was to examine the influence of conidia pigmentation, fungal development stage and genotype strain on human leucocytes oxidative response. Various A. fumigatus strains were used and the oxidative response was analysed by flow cytometry. A significant difference was observed between live- and killed-conidia. A pigmentless strain gave an important intracellular oxidative response compared with pigmented strains. But no difference was observed between strains isolated from patients with invasive aspergillosis (IA) and bronchial colonisation. The modification of healthy phagocytes' oxidative response caused by A. fumigatus components is not sufficient to explain the virulence of fungus and to predict an evolution of patients with IA.