title: Rapid tests for the detection of the Mycobacterium abscessus subsp. bolletii strain responsible for an epidemic of surgical-site infections in Brazil.
abstract: A single strain of Mycobacterium abscessus subsp. bolletii, characterised by a particular rpoB sequevar and two highly related pulsed field gel electrophoresis patterns has been responsible for a nationwide outbreak of surgical infections in Brazil since 2004. In this study, we developed molecular tests based on polymerase chain reaction restriction-enzyme analysis (PRA) and sequencing for the rapid identification of this strain. Sequences of 15 DNA regions conserved in mycobacteria were retrieved from GenBank or sequenced and analysed in silico. Single nucleotide polymorphisms specific to the epidemic strain and located in enzyme recognition sites were detected in rpoB, the 3' region of the 16S rDNA and gyrB. The three tests that were developed, i.e., PRA-rpoB, PRA-16S and gyrB sequence analysis, showed 100%, 100% and 92.31% sensitivity and 93.06%, 90.28% and 100% specificity, respectively, for the discrimination of the surgical strain from other M. abscessus subsp. bolletii isolates, including 116 isolates from 95 patients, one environmental isolate and two type strains. The results of the three tests were stable, as shown by results obtained for different isolates from the same patient. In conclusion, due to the clinical and epidemiological importance of this strain, these tests could be implemented in reference laboratories for the rapid preliminary diagnosis and epidemiological surveillance of this epidemic strain.
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP, Brasil. criskmatsumoto@yahoo.com.br
authors.forename: Cristianne Kayoko
authors.initials: CK
authors.lastname: Matsumoto
authors.forename: Erica
authors.initials: E
authors.lastname: Chimara
authors.forename: Jesus Pais
authors.initials: JP
authors.lastname: Ramos
authors.forename: Carlos Eduardo Dias
authors.initials: CE
authors.lastname: Campos
authors.forename: Paulo Cesar de Souza
authors.initials: PC
authors.lastname: Caldas
authors.forename: Karla Valeria Batista
authors.initials: KV
authors.lastname: Lima
authors.forename: Maria Luiza
authors.initials: ML
authors.lastname: Lopes
authors.forename: Rafael Silva
authors.initials: RS
authors.lastname: Duarte
authors.forename: Sylvia Cardoso
authors.initials: SC
authors.lastname: Leão
mesh_terms: D015373:Bacterial Typing Techniques / Q000379:methods; D001483:Base Sequence; D001938:Brazil; D004269:DNA, Bacterial / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D016521:Electrophoresis, Gel, Pulsed-Field; D006801:Humans; D009161:Mycobacterium / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D009164:Mycobacterium Infections / Q000453:epidemiology / Q000382:microbiology*; D017422:Sequence Analysis, DNA; D013530:Surgical Wound Infection / Q000453:epidemiology / Q000382:microbiology*
chemical_list: D004269:DNA, Bacterial; D004275:DNA, Ribosomal