Extrapulmonary infections caused by a dominant strain of Mycobacterium massiliense (Mycobacterium abscessus subspecies bolletii)  A. Cheng, Y.-C. Liu,

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Extrapulmonary infections caused by a dominant strain of Mycobacterium massiliense (Mycobacterium abscessus subspecies bolletii)  A. Cheng, Y.-C. Liu, M.-L. Chen, C.-C. Hung, Y.-T. Tsai, W.-H. Sheng, C.-H. Liao, P.-R. Hsueh, Y.-C. Chen, S.-C. Chang  Clinical Microbiology and Infection  Volume 19, Issue 10, Pages E473-E482 (October 2013) DOI: 10.1111/1469-0691.12261 Copyright © 2013 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 1 (a) Study flow processing 67 isolates of rapidly growing mycobacteria. (b) Phylogenetic trees derived from the individual and integrated concatenated hsp65, rpoB and secA sequences. The trees were constructed using minimum spanning trees (BioNumerics V.6.0, Applied Maths). All nine case isolates (red) were identified as Mycobacterium massiliense. Among the various control isolates, nine isolates were identified to be RGM other than Mycobacterium abscessus species, namely M. chelonae (n = 3, two from respiratory specimens, type strain ATCC35752), M. fortuitum (n = 3, two from surgical wound specimens, one from non-surgical wound specimens), Gordonia species (n = 2, both from respiratory specimens) and M. senaqalense (n = 1, environmental isolate). Only one M. abscessus isolate (4752) had conflicting subspecies assignments by use of the hsp65 and rpoB, with 100% identification as subsp. abscessus with the hsp65 gene, and 99.9% and 100% identification as M. massiliense by rpoB and secA1, respectively. Clinical Microbiology and Infection 2013 19, E473-E482DOI: (10.1111/1469-0691.12261) Copyright © 2013 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 2 Dendrogram of AseI pulsed-field gel electrophoretic patterns. Red line and box indicates isolates with 90% similarity by PFGE. ID, identification number of isolate. Cases of M. massiliense infections occurring post-surgery at the Children's Hospital are numbered chronologically from 1 to 9. Control specimens were obtained from patients who had never received surgery at the Children's Hospital from the following sites: SW, surgical wound; LN, lymph node; SPU, sputum; SC, subcutaneous tissue. TPE 101 refers to the major pulsotype; other, all other pulsotypes. MLSA, multilocus sequence analysis based on three genes identified the isolate to the subspecies level. Rep-PCR, repetitive sequence PCR fingerprinting was used to subgroup identical pulsotypes into repotypes with at least 80% similarity into A, B, C and so forth for M. massiliense (M. abscessus subspecies bolletii) and X, Y, Z and so forth for M. abscessus (M. abscessus subspecies abscessus). Numbers after the capital letter denote strains with at least 90% similarity by rep-PCR as in A1, A2, D1 and Y1, Y2. Clinical Microbiology and Infection 2013 19, E473-E482DOI: (10.1111/1469-0691.12261) Copyright © 2013 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 3 The skewed distribution of isolates of the major pulsotype of M. massiliense (TPE 101) among 58 isolates of M. abscessus sensu lato by tissue type, with a predominance among samples obtained from surgical wounds, subcutaneous tissues and serous fluid from body cavities, but low representation among sputum, skin surface, ocular and environmental samples. TPE 101, M. massiliense outbreak strain pulsotype A. Other types, M. abscessus and M. massiliense strains unrelated to pulsotype A. SW, surgical wound control isolates; SC, subcutaneous tissue isolates; AS/PE, ascites/pleural effusion; LN, lymph node; BLD, blood; Env., environment. Clinical Microbiology and Infection 2013 19, E473-E482DOI: (10.1111/1469-0691.12261) Copyright © 2013 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 4 Virtual gel image generated by Rep-PCR using the Diversilab Mycobacterium Strain Typing kit (Diversilab v3.4). Clinical Microbiology and Infection 2013 19, E473-E482DOI: (10.1111/1469-0691.12261) Copyright © 2013 European Society of Clinical Infectious Diseases Terms and Conditions