ST 262: a NEW SEQUENCE TYPE OF Mycobacterium abscessus SUBSP. massiliense IN PORTO ALEGRE, BRAZIL
MAIARA DOS SANTOS CARNEIRO^2,3, DAIANA LIMA-MORALES^2,3, LUCIANA DE SOUZA NUNES ^2,3, AFONSO LUÍS BARTH^2,3
2. UFRGS - Programa de Pós-graduação em Ciências Farmacêuticas (PPGCF), Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, 3. HCPA - Laboratório de Pesquisa em Resistência Bacteriana (LABRESIS), Centro de Pesquisa Experimental, Hospital de Clínicas de Porto Alegre, 4. UNIPAMPA - Universidade Federal do Pampa
lu8nunes@gmail.com

Mycobacterium abscessus complex (MABC) belongs to a group of rapidly growing mycobacteria which cause various diseases including skin and respiratory infections. This  complex  is  composed of 3  related  species : M.  abscessus subsp. abscessus , M. abscessus subsp. massiliense, and M. abscessus subsp. bolletii. MABC is one of the most drug resistant mycobacteria, being naturally resistant to many antibiotics, including the first-line tuberculostatic drugs and was related to outbreaks in several regions of Brazil. The MLST is a typing technique which can be very useful to identify sequence types (ST) associated with outbreaks. In the MLST scheme, single-copy housekeeping genes are sequenced and the result is compared to a database in order to classify the ST identified. The aim of this study was to describe a new  ST of M. abscessus subsp. massiliense. The isolate was provenient from the respiratory tract  in 2013 in Porto Alegre, Brazil. The isolate was submitted to whole genome sequencing as follows: the DNA was extracted by boiling and ultrasonic bath and a final step of purification was performed with ReliaPrep™ gDNA Tissue Miniprep System (PROMEGA). The genomic library was made with the Nextera® XT DNA Sample Preparation Kit (Illumina, San Diego, CA), followed by quantification on TapeStation (Agilent) and sequenced in the MiSeq Platform (Illumina, San Diego, CA). The genome was trimmed with Trim Galore! and assembled with SPAdes Genome Assembler. The ST was established by the in silico analysis of  seven housekeeping genes: argH (argininosuccinate lyase), cya (adenylate cyclase), gnd (6-phosphogluconate dehydrogenase), murC (UDP N-acetylmuramate-L-Ala ligase), pta (phosphate acetyltransferase), glpK ( glycerol kinase) and purH (phosphoribosylaminoimidazole carboxylase ATPase subunit). The genome was annotated on Patrick server and submitted to Center of Genomic Epidemiology MLST typing (https://cge.cbs.dtu.dk/services/MLST/). The sequence type was the result of combination of alleles argH: 11, cya: 13, glpK: 24, gnd: 10, murC: 7, pta: 11 and purH: 7, which was unique and therefore designated as a new ST. This new ST was deposited in Pausteur MLST site and was assigned as number 262.  This result indicates that there is a high probability of horizontal gene transfer among M. abcessus subsp. massiliense which can generate new STs.