Fungi identification methods

The focus of this chapter is the development of a technique often called wholecell matrix-assisted laser desorption ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) or whole-cell MALDI-TOF MS. Some groups prefer to use terms such as intact or unprocessed rather than whole, but the intended meaning is the same regardless of which word is used. As noted in the first chapter of this book, there are many different methods for the analysis of bacteria. However, for the analysis of intact or unprocessed bacteria, whole-cell MALDI-TOF MS is the most commonly used approach. This method is very rapid. MALDI-TOF MS analysis of whole cells takes only minutes because the samples can be analyzed directly after collection from a bacterial culture suspension. Direct MALDI MS analysis of fungi or viruses is similar in approach1,2 but is not covered in this chapter. MALDI-TOF MS of whole cells was developed with very rapid identification or differentiation of bacteria in mind. The name (whole cell) should not be taken to imply that the cells are literally intact or whole. Rather, it should be taken to mean that the cells that have not been treated or processed in any way specifically for the removal or isolation of any cellular components from any others. In whole-cell analysis the cells have been manipulated only as necessary to... 

Laser-induced native fluorescence is measured on a set of bio-molecules from different classes (bacteria, proteins, fungi) for excitation at 266nm and 355nm. A method of preprocessing the spectra to obtain an inherently normalized set of data is described. Class identification on the normalized data set is demonstrated. 

The application of infrared spectrophotometry to the study of bacteria and fungi has been investigated by many workers. Stevenson and Bolduan (1952) proposed the use of this method for the identification of bacteria. They found that infrared spectra do not always group species as in the Bergey classification. For example. Pseudomonas (Family II) organisms and Escherichia (Family X) had similar spectra, whereas Micrococcus rosaceous and M. pyogenes var. aureus (Genus I, Family V) had very different spectra. 

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