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112 Standard versus extended fungal culture for the detection of filamentous fungi in CF sputum samples
Posters / Journal of Cystic Fibrosis 15 (2016) S51–S120
could reflect Aspergillus infection or colonization; GM is not useful in differentiating. Further clinical information such as radiological findings and analysis of GM performance on blood is required. 110 Pitfalls in the diagnosis of Aspergillus disease in a cohort of Brazilian CF patients T.B. Aiello1 , R.M. Mauch1 , A.A.D.C. Toro1 , M.C. Pereira1 , M.T. Nolasco da Silva1 , I.A. Paschoal1 , A.F. Ribeiro1 , J.D. Ribeiro1 , C.E. Levy1 . 1 Unicamp, Campinas, Brazil Objectives: To assess the prevalence of Aspergillus disease in CF patients from a Brazilian reference center. Methods: Data of 109 patients were gathered; 55 patients had at least 1 positive Aspergillus sputum culture (prevalence 50.5%). 38 patients had at least 2 positive cultures for Aspergillus and were included in the study. Measurement of total IgE, specific Aspergillus fumigatus IgE and IgG were made for these patients. The clinical spectrum of Aspergillus disease was defined by current literature standards. Results and Discussion: Following current criteria, no patients had invasive aspergillosis; 3 patients had elevated specific IgG levels (102 mgA/L), which configures possible Aspergillus bronchitis. No patients filled the minimum criteria for ABPA. However, 3 patients had elevated specific IgE (>0.3 kUA/L), showing possible sensitization for the fungus and need to be monitored. Of these, 1 patient showed a great potential for ABP – specific IgE = 5.23 kUA/L, clinical signs of disease and good clinical outcome after treatment – despite not filling the minimum criteria for ABPA. Conclusions: Despite the prevalence of Aspergillus disease in our center being very low, a single case drew our attention, showing high specific IgE, but not total IgE levels, even the patient having food allergy with clinical manifestations. As is known, IgE is a very little fraction of the immunoglobulins and only about 50% of allergic individuals have a high total IgE level. Thus, despite our sample being small, we suggest that the requirement of total IgE for definition of ABPA, and also the necessity of fulfilment of the classic criteria at all, should be critically addressed. 111 Fungal colonization of the respiratory tract of CF patients and its impact on the respiratory function 2 E. Romanowska1 , K. Semczuk1 , A. Wojcieszek1 , H. Dmenska ´ , 1 1 A. Smorczewska-Kiljan3 , K. Dzierzanowska-Fangrat ˙ . Children’s Memorial Health Institute, Department of Clinical Microbiology and Immunology, Warsaw, Poland; 2 Chidlren’s Memorial Health Institute, Pulmonary Outpatient’s Clinic, Warsaw, Poland; 3 Chidlren’s Memorial Health Institute, Department of Pediatrics, Warsaw, Poland
Objective: To determine the impact of the respiratory tract colonisation by Aspergillus spp., Scedosporium spp. and Candida spp. in the CF patients. Methods: A total of 20 CF patients (11 girls and 9 boys; age range 19 years), treated in the Children’s Memorial Health Institute in Warsaw were included. From January 2013 to December 2015, 62 sputum samples were collected. The clinical material was inoculated on Sabouraud dextrose agar and incubated for 14 days in 35ºC. Positive yeast cultures were identified by VITEK System. Moulds were identified by macroscopic and microscopic features. Demographic, medical data (forced expiratory volume in the first second and body mass index) and microbiological findings were analysed. Results and Conclusion: Samples were negative for fungal cultures in 30% patients. The other 70% were persistently colonised with Candida spp. Among them 29% were also affected by moulds. The mean FEV1/FVC value in the group with negative cultures (79.7%) was similar to the group colonised both with Candida spp. and moulds Aspergillus spp. and Scedosporium spp. (77.2%). The mean FEV1/FVC value in patients colonised with Candida spp. only was lower: 68.3%. BMI index was comparable in all groups. Age, underweight and other factors such as bacterial coinfection were associated with FEV1 worsening. Two patients with concomitant S. prolificans, A. fumigatus, C. lusitaniae and C. dubliniensis colonisation
S79
were at the greatest risk of pulmonary exacerbations and required hospitalisation. Further studies analyzing the impact of fungal colonization on the respiratory function as well as host-pathogen response to colonisation/infection in CF patients are needed. 112 Standard versus extended fungal culture for the detection of filamentous fungi in CF sputum samples R. Ratnakumar1 , S. Oakley1 , S. Chapman1,2 , K. Jefferey1 , W.G. Flight1 . 1 Oxford University Hospitals NHS Foundation Trust, Oxford, United Kingdom; 2 Oxford University, Oxford, United Kingdom Background: Filamentous fungi such as Aspergillus are frequently identified in the sputum of patients with cystic fibrosis (CF). Diagnosis of fungal infection in CF is challenging, and optimal diagnostic methods unclear. We compared standard (48 hour) with extended (3 week) fungal culture of sputum from adults with CF. Methods: Paired CF sputum samples processed with both 48-hour culture on Sabouraud agar and extended culture on Sabouraud slopes from Oct 2014 to Sept 2015 were identified retrospectively and compared to assess species cultured and comparative pick-up rate of fungi. Patients with prior lung transplantation were excluded. Results: 994 sputum samples were identified from 95 patients over the study period, of which 538 had paired standard and extended fungal culture results. 174/538 (32.3%) samples were positive for a filamentous fungus. Extended culture was positive in 156 (89.7%), standard culture in 118(67.8%) and both were positive in 100 (57.5%). Standard culture had a sensitivity of 64.1% and specificity of 95.3%. 53 patients (56%) had ≥1 sample positive for a fungus. Aspergillus fumigatus was isolated from 51 patients with other species of Aspergillus found in 5 patients. Rasamsonia argillacea, Exophiala phialophora and Scedosporium apiospermum were detected in 1 patient each. Patients with ≥1 positive sample had higher Aspergillus precipitins [mean 2.5 (SD1.9) vs 0.8 (1.3) lines, p < 0.001] and were more likely to have had NTM isolated previously (49% vs 26%, p = 0.03). Conclusions: Filamentous fungi are frequently detected in CF sputum, A. fumigatus being the most common species. Extended fungal culture increases fungal detection rate. 113 Evaluation of new approach for diagnosis of fungal rhinosinusitis in patients with cystic fibrosis A. Barac1 , P. Minic2,3 , V. Tomic Spiric4,5 , E. Aleksic6 , V. Arsic Arsenijevic1 . 1 Faculty of Medicine, University of Belgrade, Institute of Microbiology and Immunology, Belgrade, Serbia; 2 Institute of Mother and Child, Belgrade, Serbia; 3 Faculty of Medicine, University of Belgrade, Belgrade, Serbia; 4 Clinic for Allergology and Clinical Immunology, Clinical Center of Serbia, Belgrade, Serbia; 5 Faculty of Medicine University of Belgrade, Belgrade, Serbia; 6 Faculty of Stomatology, Business Academy of Novi Sad, Pancevo, Pancevo, Serbia Background: Although the pathogenesis of cystic fibrosis (CF) and fungal rhinosinusitis (FRS) have been widely studied, the relationship of the mycobiome of the sinonasal mucosa and development of FRS is not yet clear. The aim of our study was to evaluate new approach for diagnosis of FRS in patients with CF in order to prevent development of invasive respiratory fungal diseases. Methodology: A prospective study with 42 patients was conducted in the Faculty of Medicine, University of Belgrade. Patients were divided into two groups depending on the presence of CF: patients with CF (CF+) and patients without CF (CF−). After mycological analyses (using inhalation with PARI-SINUS nebulizer and sampling) patients were divided in groups: 1. patients with positive fungal finding and CF (FRS group) and 2. patients without CF and positive fungal finding (non-FRS group). Study design included: 1. rhinologic observation and 2. mycological finding of sinonasal aspirate. Results: 6.4% of patients from CF group had positive fungal finding on nasal mucosa and could be considered as FRS. Patients from FRS group
could reflect Aspergillus infection or colonization; GM is not useful in differentiating. Further clinical information such as radiological findings and analysis of GM performance on blood is required. 110 Pitfalls in the diagnosis of Aspergillus disease in a cohort of Brazilian CF patients T.B. Aiello1 , R.M. Mauch1 , A.A.D.C. Toro1 , M.C. Pereira1 , M.T. Nolasco da Silva1 , I.A. Paschoal1 , A.F. Ribeiro1 , J.D. Ribeiro1 , C.E. Levy1 . 1 Unicamp, Campinas, Brazil Objectives: To assess the prevalence of Aspergillus disease in CF patients from a Brazilian reference center. Methods: Data of 109 patients were gathered; 55 patients had at least 1 positive Aspergillus sputum culture (prevalence 50.5%). 38 patients had at least 2 positive cultures for Aspergillus and were included in the study. Measurement of total IgE, specific Aspergillus fumigatus IgE and IgG were made for these patients. The clinical spectrum of Aspergillus disease was defined by current literature standards. Results and Discussion: Following current criteria, no patients had invasive aspergillosis; 3 patients had elevated specific IgG levels (102 mgA/L), which configures possible Aspergillus bronchitis. No patients filled the minimum criteria for ABPA. However, 3 patients had elevated specific IgE (>0.3 kUA/L), showing possible sensitization for the fungus and need to be monitored. Of these, 1 patient showed a great potential for ABP – specific IgE = 5.23 kUA/L, clinical signs of disease and good clinical outcome after treatment – despite not filling the minimum criteria for ABPA. Conclusions: Despite the prevalence of Aspergillus disease in our center being very low, a single case drew our attention, showing high specific IgE, but not total IgE levels, even the patient having food allergy with clinical manifestations. As is known, IgE is a very little fraction of the immunoglobulins and only about 50% of allergic individuals have a high total IgE level. Thus, despite our sample being small, we suggest that the requirement of total IgE for definition of ABPA, and also the necessity of fulfilment of the classic criteria at all, should be critically addressed. 111 Fungal colonization of the respiratory tract of CF patients and its impact on the respiratory function 2 E. Romanowska1 , K. Semczuk1 , A. Wojcieszek1 , H. Dmenska ´ , 1 1 A. Smorczewska-Kiljan3 , K. Dzierzanowska-Fangrat ˙ . Children’s Memorial Health Institute, Department of Clinical Microbiology and Immunology, Warsaw, Poland; 2 Chidlren’s Memorial Health Institute, Pulmonary Outpatient’s Clinic, Warsaw, Poland; 3 Chidlren’s Memorial Health Institute, Department of Pediatrics, Warsaw, Poland
Objective: To determine the impact of the respiratory tract colonisation by Aspergillus spp., Scedosporium spp. and Candida spp. in the CF patients. Methods: A total of 20 CF patients (11 girls and 9 boys; age range 19 years), treated in the Children’s Memorial Health Institute in Warsaw were included. From January 2013 to December 2015, 62 sputum samples were collected. The clinical material was inoculated on Sabouraud dextrose agar and incubated for 14 days in 35ºC. Positive yeast cultures were identified by VITEK System. Moulds were identified by macroscopic and microscopic features. Demographic, medical data (forced expiratory volume in the first second and body mass index) and microbiological findings were analysed. Results and Conclusion: Samples were negative for fungal cultures in 30% patients. The other 70% were persistently colonised with Candida spp. Among them 29% were also affected by moulds. The mean FEV1/FVC value in the group with negative cultures (79.7%) was similar to the group colonised both with Candida spp. and moulds Aspergillus spp. and Scedosporium spp. (77.2%). The mean FEV1/FVC value in patients colonised with Candida spp. only was lower: 68.3%. BMI index was comparable in all groups. Age, underweight and other factors such as bacterial coinfection were associated with FEV1 worsening. Two patients with concomitant S. prolificans, A. fumigatus, C. lusitaniae and C. dubliniensis colonisation
S79
were at the greatest risk of pulmonary exacerbations and required hospitalisation. Further studies analyzing the impact of fungal colonization on the respiratory function as well as host-pathogen response to colonisation/infection in CF patients are needed. 112 Standard versus extended fungal culture for the detection of filamentous fungi in CF sputum samples R. Ratnakumar1 , S. Oakley1 , S. Chapman1,2 , K. Jefferey1 , W.G. Flight1 . 1 Oxford University Hospitals NHS Foundation Trust, Oxford, United Kingdom; 2 Oxford University, Oxford, United Kingdom Background: Filamentous fungi such as Aspergillus are frequently identified in the sputum of patients with cystic fibrosis (CF). Diagnosis of fungal infection in CF is challenging, and optimal diagnostic methods unclear. We compared standard (48 hour) with extended (3 week) fungal culture of sputum from adults with CF. Methods: Paired CF sputum samples processed with both 48-hour culture on Sabouraud agar and extended culture on Sabouraud slopes from Oct 2014 to Sept 2015 were identified retrospectively and compared to assess species cultured and comparative pick-up rate of fungi. Patients with prior lung transplantation were excluded. Results: 994 sputum samples were identified from 95 patients over the study period, of which 538 had paired standard and extended fungal culture results. 174/538 (32.3%) samples were positive for a filamentous fungus. Extended culture was positive in 156 (89.7%), standard culture in 118(67.8%) and both were positive in 100 (57.5%). Standard culture had a sensitivity of 64.1% and specificity of 95.3%. 53 patients (56%) had ≥1 sample positive for a fungus. Aspergillus fumigatus was isolated from 51 patients with other species of Aspergillus found in 5 patients. Rasamsonia argillacea, Exophiala phialophora and Scedosporium apiospermum were detected in 1 patient each. Patients with ≥1 positive sample had higher Aspergillus precipitins [mean 2.5 (SD1.9) vs 0.8 (1.3) lines, p < 0.001] and were more likely to have had NTM isolated previously (49% vs 26%, p = 0.03). Conclusions: Filamentous fungi are frequently detected in CF sputum, A. fumigatus being the most common species. Extended fungal culture increases fungal detection rate. 113 Evaluation of new approach for diagnosis of fungal rhinosinusitis in patients with cystic fibrosis A. Barac1 , P. Minic2,3 , V. Tomic Spiric4,5 , E. Aleksic6 , V. Arsic Arsenijevic1 . 1 Faculty of Medicine, University of Belgrade, Institute of Microbiology and Immunology, Belgrade, Serbia; 2 Institute of Mother and Child, Belgrade, Serbia; 3 Faculty of Medicine, University of Belgrade, Belgrade, Serbia; 4 Clinic for Allergology and Clinical Immunology, Clinical Center of Serbia, Belgrade, Serbia; 5 Faculty of Medicine University of Belgrade, Belgrade, Serbia; 6 Faculty of Stomatology, Business Academy of Novi Sad, Pancevo, Pancevo, Serbia Background: Although the pathogenesis of cystic fibrosis (CF) and fungal rhinosinusitis (FRS) have been widely studied, the relationship of the mycobiome of the sinonasal mucosa and development of FRS is not yet clear. The aim of our study was to evaluate new approach for diagnosis of FRS in patients with CF in order to prevent development of invasive respiratory fungal diseases. Methodology: A prospective study with 42 patients was conducted in the Faculty of Medicine, University of Belgrade. Patients were divided into two groups depending on the presence of CF: patients with CF (CF+) and patients without CF (CF−). After mycological analyses (using inhalation with PARI-SINUS nebulizer and sampling) patients were divided in groups: 1. patients with positive fungal finding and CF (FRS group) and 2. patients without CF and positive fungal finding (non-FRS group). Study design included: 1. rhinologic observation and 2. mycological finding of sinonasal aspirate. Results: 6.4% of patients from CF group had positive fungal finding on nasal mucosa and could be considered as FRS. Patients from FRS group