Evaluation of phenotypical and genotypical methods for the identification and typing of Stenotrophomonas maltophilia isolated from a pharmaceutical facility

Author:

de Miranda Rebeca Vitória da Silva Lage12ORCID,Monteiro Giovanna Merrelho1,da Costa Luciana Veloso1,dos Santos Milena Cristina Silva1,dos Reis Cristhiane Moura Falavina1,Braga Lygia Maria Paulo da Silva1,Forsythe Stephen James3,Villas Bôas Maria Helena Simões2,Brandão Marcelo Luiz Lima1ORCID

Affiliation:

1. Laboratory of Microbiological Control, Bio-Manguinhos/Fiocruz , CEP 21040-900, Rio de Janeiro , Brazil

2. Laboratory of Microbiology of Food and Sanitizes, INCQS/Fiocruz , CEP 21040-900, Rio de Janeiro , Brazil

3. Foodmicrobe.com, Adams Hill , Keyworth, Nottinghamshire NG12 5GY , United Kingdom

Abstract

Abstract Aims Evaluate methods for identification and typing of Stenotrophomonas maltophilia isolated from a pharmaceutical facility. Methods and results From 270 S. maltophilia strains identified by VITEK®2, 40 were selected and submitted to MALDI TOF-MS, 16S and 23S rRNA gene analysis, enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR), and an antimicrobial susceptibility profile. 16S rRNA sequencing was able to identify 39 (97.5%) strains as Stenotrophomonas spp. and one (2.5%) as Luteimonas huabeiensis. MALDI TOF-MS identified 37 (92.5%) strains as S. maltophilia, and three (7.5%) were not identified. PCR targeting 23S rRNA yielded a positive result for 39 (97.5%) strains. However, after sequencing, two strains were identified as Stenotrophomonas rhizophila, showing false-positive results. The confirmed S. maltophilia strains (n = 37) showed 35 distinct ERIC-PCR profiles and exhibited sensitivity to minocycline and levofloxacin, and six (16.3%) showed intermediate resistance to sulfamethoxazole-trimethoprim. Conclusion Matrix-assisted laser desorption lonization-time of flight mass spectrometry (MALDI-TOF MS) was a satisfactory methodology for the identification of S. maltophilia, but expansion of the database is necessary for the identification of other species. 16S rDNA sequencing showed low resolution for Stenotrophomonas species differentiation. PCR targeting 23S rRNA could not differentiate S. maltophilia from S. rhizophila. ERIC-PCR was shown to be a useful tool for the microbial source tracking of S. maltophilia.

Funder

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil

Publisher

Oxford University Press (OUP)

Subject

Applied Microbiology and Biotechnology,General Medicine,Biotechnology

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