FT-IR microspectroscopy in rapid identification of bacteria in pure and mixed culture

Inglid Fontoura; Ricardo Belo; Kumiko Sakane; Maria Angélica Gargione Cardoso; Sônia Khouri; Mituo Uehara; Leandro Raniero; Airton A. Martin

doi:10.1117/12.841918

11 February 2010 FT-IR microspectroscopy in rapid identification of bacteria in pure and mixed culture

Inglid Fontoura, Ricardo Belo, Kumiko Sakane, Maria Angélica Gargione Cardoso, Sônia Khouri, Mituo Uehara, Leandro Raniero, Airton A. Martin

Author Affiliations +

Proceedings Volume 7560, Biomedical Vibrational Spectroscopy IV: Advances in Research and Industry; 75600T (2010) https://doi.org/10.1117/12.841918
Event: SPIE BiOS, 2010, San Francisco, California, United States

Abstract

In recent years FT-IR microspectroscopy has been developed for microbiology analysis and applied successfully in pure cultures of microorganisms to rapidly identify strains of bacteria, yeasts and fungi. The investigation and characterization of microorganism mixed cultures is also of growing importance, especially in hospitals where it is common to poly-microbial infections. In this work, the rapid identification of bacteria in pure and mixed cultures was studied. The bacteria were obtained from the Institute Oswaldo Cruz culture collection at Brazil. Escherichia coli ATCC 10799 and Staphylococcus aureus ATCC 14456 were analyzed, 3 inoculations were examined in triplicate: Escherichia coli, Staphylococcus aureus and a mixed culture of them. The inoculations were prepared according to McFarland 0.5, incubated at 37 ° C for 6 hours, diluted in saline, placed in the CaF2 window and store for one hour at 50°C to obtain thin film. The measurement was performed by Spectrum Spotlight 400 (Perkin-Elmer) equipment in the range of 4000-900 cm^-1, with 32 scans using a transmittance technique with point and image modes. The data were processed (baseline, normalization, calculation of first derivate followed by smoothing with 9 point using a Savitzky-Golay algorithm) and a cluster analysis were done by Ward's algorithm and an excellent discrimination between pure and mixed culture was obtained. Our preliminary results indicate that the FT-IR microspectroscopy associated with cluster analysis can be used to discriminate between pure and mixed culture.

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Inglid Fontoura, Ricardo Belo, Kumiko Sakane, Maria Angélica Gargione Cardoso, Sônia Khouri, Mituo Uehara, Leandro Raniero, and Airton A. Martin "FT-IR microspectroscopy in rapid identification of bacteria in pure and mixed culture", Proc. SPIE 7560, Biomedical Vibrational Spectroscopy IV: Advances in Research and Industry, 75600T (11 February 2010); https://doi.org/10.1117/12.841918

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