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106 Evaluation of the use of selective Burkholderia cepacia media for detection of Mycobacterium abscessus
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Posters / Journal of Cystic Fibrosis 15 (2016) S51–S120
antibiotic suppressive therapy should be considered especially for those patients considered for lung transplantation. Sort and duration of long term antibiotic suppressive therapy need to be assessed with prospective cohort studies.
Conclusion: The selective B. cepacia agar method is a sensitive, costeffective method for monitoring M. abscessus colonisation in patients with chronic infection. In this instance it was not a reliable method for detection of new infection compared to the LJ slope.
105 Clinical evaluation of cystic fibrosis patients with Mycobacterium abscessus infection 3 N. Emiralioglu1 , G.D. Tugcu1 , B. Er2 , Z. Saribas3 , B. Sener ¸ , E. Yalcın ¸ 1, 1 1 ¨ celik ¸ . Hacettepe University Faculty of D. Dogru1 , N. Kiper1 , U. Oz Medicine, Pediatric Pulmonology, Ankara, Turkey; 2 Hacettepe University Faculty of Medicine, Adult Pulmonology, Ankara, Turkey; 3 Hacettepe University Faculty of Medicine, Medical Microbiology, Ankara, Turkey
107 Persistence of livestock-associated MRSA in the airways of CF patients
Objectives: Nontuberculous mycobacteria(NTM) are increasingly implicated as a clinically important infection in cystic fibrosis(CF). The majority of patients with CF with a first positive NTM culture does not require treatment, and one fourth of subjects clear their sputum spontaneously. Here we aimed to assess M. abscessus infection prevalance and associated clinical, microbiological features and outcomes in patients with CF. Methods: We retrospectively reviewed the patients with CF who had M. abscessus positive culture and followed at Hacettepe University Pediatric Pulmonology Department between years of 2013 and 2015. We evaluated clinical, microbiological outcomes of 7 patients with at least one positive culture for M. abscessus. Results: The age of the patients ranged from 12 years to 40 years (mean 22.7 years). Their mean age at the diagnosis of first positive culture for M.abscessus was 26.6 years and there were three male patients. Six patients had chronic Pseudomonas aeruginosa infection, 3 patients had chronic Staphylococcus aereus infection. NTM was eradicated with meropenem, clarithromycin and amikacin therapy for 6 months in one of them, but it could not be eradicated in two patients despite 13 months and 4 years treatment duration. The other 3 patients had sputum conversion after the first culture without any treatment. Conclusion: Here we showed that nearly half of our patients with CF with a first positive NTM culture did not require treatment and cleared their sputum spontaneously. Assessment of treatment success should include clinical evaluation, repeated sputum cultures, spirometry and chest imaging. 106 Evaluation of the use of selective Burkholderia cepacia media for detection of Mycobacterium abscessus K. Stickland1 , C. Auckland1 , A. Chapman1 , C. Sheldon2 . 1 Royal Devon and Exeter NHS Foundation Trust, Microbiology, Exeter, United Kingdom; 2 Royal Devon and Exeter NHS Foundation Trust, Respiratory Medicine, Exeter, United Kingdom Objectives: To assess the sensitivity of using Burkholderia cepacia media to detect Mycobacterium abscessus in routine sputum cultures from Cystic Fibrosis (CF) patients. Methods: Selective B. cepacia agar was inoculated with 100 ml of neat, homogenised sputum, examined after 24 & 48 hrs incubation at 37o C, and re-incubated at 30o C for 3 days as part of routine CF culture for Burkolderia spp. Plates were re-incubated at 30o C and examined for Mycobacteria spp. after a further 9 days (14 days total). Suspicious growth was stained for AFBs using the ZN technique and identified in the reference laboratory. TB culture, including an auramine phenol stain, was performed if requested. TB culture included decontamination with 4% NaOH, neutralisation with up to 20 ml sterile water and centrifugation at 3000rpm. Supernatant was discarded and 2 drops of the deposit inoculated onto a Low-Acid pyruvate and a Low-Acid glycerol LJ-slope, incubated at 37o C for up to 12 weeks. Results: 304 samples were cultured using B. cepacia agar, 29 of which had parallel TB culture. 11 samples (4 patients) were positive for AFB on B.cepacia agar after 14 days, 3 of these (2 patients) were also cultured for Mycobacteria using LJ slopes and were also positive at 14 days. 4 patients with M.abscessus on B.cepacia media had previous isolates. 2 samples in a newly infected patient were negative on selective B. cepacia agar but positive on LJ slope.
1 S. van Alen1 , B. Ballhausen1 , M. Bach1 , A. Dubbers ¨ , J. Grosse-Onnebrink1 , 2 2 P. Kuster ¨ , H. Schultingkemper ¨ , K. Becker1 , B.C. Kahl3 . 1 University Hospital M¨ unster, M¨ unster, Germany; 2 Clemenshospital M¨ unster, M¨ unster, unster, Medical Microbiology, M¨ unster, Germany; 3 University Hospital M¨ Germany
Objectives: One of the most prevalent pathogens recovered from CF airways is Staphylococcus aureus. There is no information about the prevalence and persistence of livestock-associated (LA) MRSA in CF. Methods: We determined the prevalence of MRSA in two certified CF centers in Munster. ¨ MRSA strains were molecular typed by spa-typing. Sequential LA-MRSA isolates from individual patients were subjected to DNA microarray analysis (IdentiBAC Microarray, Alere Technologies GmbH, Jena, Germany). Results: MRSA was recovered from 29 of 189 (15%) CF patients. LAMRSA was identified in 9 patients (31%), all belonging to CC398 (in 4 patients t034, in 3 patients t011, in one patient t108 and t2011). Thirteen LA-MRSA isolates from 3 patients (n = 3, t108; n = 8, n = 2, all t034) were subjected to DNA microarray analysis. All isolates carried the methicillin- and tetracyclin-resistance genes mecA and tetM, while the erythromycin resistance gene ermA was detected in all isolates of patients 1 and 2, ermC in both isolates of patient 3, tetK in all isolates of patients 1 and 3 and the aminoglycoside resistance gene aadD in 4 of 8 isolates of patient 2 and in both isolates of patient 3. lukX was present in 10/13 and lukY in 9/13 isolates. All isolates belonged to capsule type 5. While the proteases sspA, B and P, the biofilm genes icaA, C and D were detected in all isolates, the protease aur was missing in one, the adhesin fib was present in the first isolate of patient 1 only and sdrD in 10/13 isolates. Conclusion: LA-MRSA can be isolated from the airways of CF patients and can colonize/infect the airways for extended periods. 109 An evaluation of the performance of a galactomannan (GM) assay in aiding the diagnosis of allergic broncho-pulmonary aspergillosis (ABPA) in cystic fibrosis (CF) patients R. Dhillon1 , L. Speight2 , J. Duckers2 , P.L. White1 . 1 Public Health Wales, Microbiology, Cardiff, United Kingdom; 2 University Hospital Llandough, All Wales Adult CF Centre, Cardiff, United Kingdom Objectives: Evaluation of the role of GM in the diagnosis of ABPA vs non-ABPA states in CF patients. Methods: Prospective data was collected on patients (n = 35) who had sputum samples sent for GM analysis. Total and Aspergillusspecific IgE, Aspergillus IgG, eosinophil counts were accrued. Clinical information, including lung function, steroid and/or antifungal use was collected. ABPA cases and control patients were defined using the CF foundation consensus criteria. Performance of GM analysis was determined alongside regression analysis to identify any association between GM and clinical markers of ABPA. ROC analysis was performed to determine optimal GM threshold when suspecting ABPA. Results: There were 4 cases of ABPA, 4 cases of Aspergillus infection,15 patients with Aspergillus colonisation and 10 control patients. The sensitivity and specificity of GM analysis when testing sputum was 100% and 20%, respectively. All patients with Aspergillus infection were positive, as were 13/15 patients with colonisation. Regression analysis identified a single significant association between GM index and serum Aspergillus IgG concentration (p = 0.05). ROC analysis generated AUC:0.53 with a GM threshold of 5.5 appearing optimal, but this could not be used to differentiate colonisation Conclusion: The use of GM analysis on sputum to aid in the diagnosis of ABPA in CF is limited. If GM is consistently negative, ABPA is unlikely. GM sputum positivity was associated with serum Aspergillus IgG but this
Posters / Journal of Cystic Fibrosis 15 (2016) S51–S120
antibiotic suppressive therapy should be considered especially for those patients considered for lung transplantation. Sort and duration of long term antibiotic suppressive therapy need to be assessed with prospective cohort studies.
Conclusion: The selective B. cepacia agar method is a sensitive, costeffective method for monitoring M. abscessus colonisation in patients with chronic infection. In this instance it was not a reliable method for detection of new infection compared to the LJ slope.
105 Clinical evaluation of cystic fibrosis patients with Mycobacterium abscessus infection 3 N. Emiralioglu1 , G.D. Tugcu1 , B. Er2 , Z. Saribas3 , B. Sener ¸ , E. Yalcın ¸ 1, 1 1 ¨ celik ¸ . Hacettepe University Faculty of D. Dogru1 , N. Kiper1 , U. Oz Medicine, Pediatric Pulmonology, Ankara, Turkey; 2 Hacettepe University Faculty of Medicine, Adult Pulmonology, Ankara, Turkey; 3 Hacettepe University Faculty of Medicine, Medical Microbiology, Ankara, Turkey
107 Persistence of livestock-associated MRSA in the airways of CF patients
Objectives: Nontuberculous mycobacteria(NTM) are increasingly implicated as a clinically important infection in cystic fibrosis(CF). The majority of patients with CF with a first positive NTM culture does not require treatment, and one fourth of subjects clear their sputum spontaneously. Here we aimed to assess M. abscessus infection prevalance and associated clinical, microbiological features and outcomes in patients with CF. Methods: We retrospectively reviewed the patients with CF who had M. abscessus positive culture and followed at Hacettepe University Pediatric Pulmonology Department between years of 2013 and 2015. We evaluated clinical, microbiological outcomes of 7 patients with at least one positive culture for M. abscessus. Results: The age of the patients ranged from 12 years to 40 years (mean 22.7 years). Their mean age at the diagnosis of first positive culture for M.abscessus was 26.6 years and there were three male patients. Six patients had chronic Pseudomonas aeruginosa infection, 3 patients had chronic Staphylococcus aereus infection. NTM was eradicated with meropenem, clarithromycin and amikacin therapy for 6 months in one of them, but it could not be eradicated in two patients despite 13 months and 4 years treatment duration. The other 3 patients had sputum conversion after the first culture without any treatment. Conclusion: Here we showed that nearly half of our patients with CF with a first positive NTM culture did not require treatment and cleared their sputum spontaneously. Assessment of treatment success should include clinical evaluation, repeated sputum cultures, spirometry and chest imaging. 106 Evaluation of the use of selective Burkholderia cepacia media for detection of Mycobacterium abscessus K. Stickland1 , C. Auckland1 , A. Chapman1 , C. Sheldon2 . 1 Royal Devon and Exeter NHS Foundation Trust, Microbiology, Exeter, United Kingdom; 2 Royal Devon and Exeter NHS Foundation Trust, Respiratory Medicine, Exeter, United Kingdom Objectives: To assess the sensitivity of using Burkholderia cepacia media to detect Mycobacterium abscessus in routine sputum cultures from Cystic Fibrosis (CF) patients. Methods: Selective B. cepacia agar was inoculated with 100 ml of neat, homogenised sputum, examined after 24 & 48 hrs incubation at 37o C, and re-incubated at 30o C for 3 days as part of routine CF culture for Burkolderia spp. Plates were re-incubated at 30o C and examined for Mycobacteria spp. after a further 9 days (14 days total). Suspicious growth was stained for AFBs using the ZN technique and identified in the reference laboratory. TB culture, including an auramine phenol stain, was performed if requested. TB culture included decontamination with 4% NaOH, neutralisation with up to 20 ml sterile water and centrifugation at 3000rpm. Supernatant was discarded and 2 drops of the deposit inoculated onto a Low-Acid pyruvate and a Low-Acid glycerol LJ-slope, incubated at 37o C for up to 12 weeks. Results: 304 samples were cultured using B. cepacia agar, 29 of which had parallel TB culture. 11 samples (4 patients) were positive for AFB on B.cepacia agar after 14 days, 3 of these (2 patients) were also cultured for Mycobacteria using LJ slopes and were also positive at 14 days. 4 patients with M.abscessus on B.cepacia media had previous isolates. 2 samples in a newly infected patient were negative on selective B. cepacia agar but positive on LJ slope.
1 S. van Alen1 , B. Ballhausen1 , M. Bach1 , A. Dubbers ¨ , J. Grosse-Onnebrink1 , 2 2 P. Kuster ¨ , H. Schultingkemper ¨ , K. Becker1 , B.C. Kahl3 . 1 University Hospital M¨ unster, M¨ unster, Germany; 2 Clemenshospital M¨ unster, M¨ unster, unster, Medical Microbiology, M¨ unster, Germany; 3 University Hospital M¨ Germany
Objectives: One of the most prevalent pathogens recovered from CF airways is Staphylococcus aureus. There is no information about the prevalence and persistence of livestock-associated (LA) MRSA in CF. Methods: We determined the prevalence of MRSA in two certified CF centers in Munster. ¨ MRSA strains were molecular typed by spa-typing. Sequential LA-MRSA isolates from individual patients were subjected to DNA microarray analysis (IdentiBAC Microarray, Alere Technologies GmbH, Jena, Germany). Results: MRSA was recovered from 29 of 189 (15%) CF patients. LAMRSA was identified in 9 patients (31%), all belonging to CC398 (in 4 patients t034, in 3 patients t011, in one patient t108 and t2011). Thirteen LA-MRSA isolates from 3 patients (n = 3, t108; n = 8, n = 2, all t034) were subjected to DNA microarray analysis. All isolates carried the methicillin- and tetracyclin-resistance genes mecA and tetM, while the erythromycin resistance gene ermA was detected in all isolates of patients 1 and 2, ermC in both isolates of patient 3, tetK in all isolates of patients 1 and 3 and the aminoglycoside resistance gene aadD in 4 of 8 isolates of patient 2 and in both isolates of patient 3. lukX was present in 10/13 and lukY in 9/13 isolates. All isolates belonged to capsule type 5. While the proteases sspA, B and P, the biofilm genes icaA, C and D were detected in all isolates, the protease aur was missing in one, the adhesin fib was present in the first isolate of patient 1 only and sdrD in 10/13 isolates. Conclusion: LA-MRSA can be isolated from the airways of CF patients and can colonize/infect the airways for extended periods. 109 An evaluation of the performance of a galactomannan (GM) assay in aiding the diagnosis of allergic broncho-pulmonary aspergillosis (ABPA) in cystic fibrosis (CF) patients R. Dhillon1 , L. Speight2 , J. Duckers2 , P.L. White1 . 1 Public Health Wales, Microbiology, Cardiff, United Kingdom; 2 University Hospital Llandough, All Wales Adult CF Centre, Cardiff, United Kingdom Objectives: Evaluation of the role of GM in the diagnosis of ABPA vs non-ABPA states in CF patients. Methods: Prospective data was collected on patients (n = 35) who had sputum samples sent for GM analysis. Total and Aspergillusspecific IgE, Aspergillus IgG, eosinophil counts were accrued. Clinical information, including lung function, steroid and/or antifungal use was collected. ABPA cases and control patients were defined using the CF foundation consensus criteria. Performance of GM analysis was determined alongside regression analysis to identify any association between GM and clinical markers of ABPA. ROC analysis was performed to determine optimal GM threshold when suspecting ABPA. Results: There were 4 cases of ABPA, 4 cases of Aspergillus infection,15 patients with Aspergillus colonisation and 10 control patients. The sensitivity and specificity of GM analysis when testing sputum was 100% and 20%, respectively. All patients with Aspergillus infection were positive, as were 13/15 patients with colonisation. Regression analysis identified a single significant association between GM index and serum Aspergillus IgG concentration (p = 0.05). ROC analysis generated AUC:0.53 with a GM threshold of 5.5 appearing optimal, but this could not be used to differentiate colonisation Conclusion: The use of GM analysis on sputum to aid in the diagnosis of ABPA in CF is limited. If GM is consistently negative, ABPA is unlikely. GM sputum positivity was associated with serum Aspergillus IgG but this