title: Enterobacterial repetitive intergenic consensus PCR is a useful tool for typing Mycobacterium chelonae and Mycobacterium abscessus isolates.
abstract: Outbreaks of rapidly growing mycobacterium (RGM) infections are increasingly being reported worldwide. Information about genetic relatedness of isolates obtained during outbreaks can provide opportunities for prompt intervention. Pulsed-field gel electrophoresis (PFGE) is expensive, time consuming, and labor intensive. Other than that, Mycobacterium abscessus isolates can suffer DNA degradation during electrophoresis. Polymerase chain reaction (PCR)-based methods are cheaper, faster, and easier to perform, but discriminatory power varies depending on the primer used. In this study, we tested the competence of enterobacterial repetitive intergenic consensus (ERIC) PCR in comparison with PFGE to distinguish unrelated isolates (24 Mycobacterium chelonae and 24 M. abscessus) obtained from human and/or environmental samples and to group 56 isolates from 6 outbreaks confirmed epidemiologically, caused by M. chelonae and M. abscessus after ophthalmologic refractive surgery and mesotherapy. Enterobacterial repetitive intergenic consensus PCR presented discriminatory power, calculated using Simpson's index of diversity, of 0.989 for M. abscessus and 0.975 for M. chelonae and grouped outbreak isolates in distinct groups showing epidemiologic concordance. Pulsed-field gel electrophoresis also grouped outbreak isolates and presented discriminatory power of 0.972 and 0.993 for M. abscessus and M. chelonae, respectively. DNA from 8 (22%) of 36 M. abscessus isolates analyzed showed degradation during electrophoresis. Compared with PFGE and epidemiologic information as the gold standard, ERIC PCR is a simple, high throughput, affordable, reproducible, and discriminatory molecular typing method for inference of genetic relatedness of RGMs of the M. chelonae-abscessus group.
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo-Escola Paulista de Medicina, Rua Botucatu, 862-3 andar-04023-062, São Paulo, Brazil.
authors.forename: Jorge Luiz Mello
authors.initials: JL
authors.lastname: Sampaio
authors.forename: Cristina
authors.initials: C
authors.lastname: Viana-Niero
authors.forename: Denise
authors.initials: D
authors.lastname: de Freitas
authors.forename: Ana Luisa
authors.initials: AL
authors.lastname: Höfling-Lima
authors.forename: Sylvia Cardoso
authors.initials: SC
authors.lastname: Leão
mesh_terms: D001431:Bacteriological Techniques; D004269:DNA, Bacterial / Q000235:genetics; D021901:DNA, Intergenic / Q000235:genetics*; D004196:Disease Outbreaks; D016521:Electrophoresis, Gel, Pulsed-Field; D015818:Eye Infections, Bacterial / Q000175:diagnosis / Q000453:epidemiology / Q000382:microbiology; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D020071:Interspersed Repetitive Sequences / Q000235:genetics*; D009161:Mycobacterium / Q000145:classification* / Q000235:genetics*; D009164:Mycobacterium Infections / Q000175:diagnosis / Q000453:epidemiology / Q000382:microbiology*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D017192:Skin Diseases, Bacterial / Q000175:diagnosis / Q000453:epidemiology / Q000382:microbiology
chemical_list: D004269:DNA, Bacterial; D021901:DNA, Intergenic