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Molecular Identification and characterization of filamentous fungi and yeasts isolated in a pharmaceutical industry environment

Luciana Nobre Leite, Felipe José Nobre Lelis, Mauro Aparecido de Sousa Xavier, Josiane dos Santos, Leia Cardoso, Frederico Santos Barbosa, Rosimar Fonseca dos Santos, Soraia Aparecida Maia Dias, Alessandra Rejane Ericsson de Oliveira Xavier.

Cited by 4 Articles

The microbiological monitoring of aseptic areas in pharmaceutical industries is an important procedure for the evaluation of the effectiveness of contamination control measures in these areas. Once the permitted microbiological level has been exceeded, the microbial contaminant should be identified, and actions to eliminate this agent should be adopted. The objective of this study was to identify filamentous fungi and yeasts isolated from the industrial environment by the MALDI-TOF MS proteomic approach, as well as to analyze the PCR-RFLP capacity of the fungal universal region in discriminating between and intraspecies differences in these isolates. The MALDI-TOF method was able to identify 100% of the samples to the genus level; however, only 68.42% of the samples were identified to the species level. The most common microorganisms among the samples were Candida guilliermondii, Penicillium spp., Rhodosporidium toruloides, and Aspergillus species. PCR-RFLP analysis of the e universal fungal region revealed a heterogeneous profile of the analyzed samples. Isolates of the same species, collected from different sampling points, presented the same restriction profile. The MALDI-TOF and PCR-RFLP methods of the universal fungal region were able respectively to identify, characterize the genetic diversity and to differentiate interspecifically the yeast and fungi samples analyzed in this study.

Key words: MALDI-TOF, PCR-RFLP, rRNA, yeast, fungi, environmental monitoring

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