Abstract

To apply TMAH-based Py-MS to a field biological detection system for real-time classification of cell-type, reproducible patterns of the TMAH-based Py-MS spectra was known as a critical factor for classification but was seriously disturbed by quantity of cells injected into pyro-tube. This factor is an exterior variable that could not be complemented by improving the performance of the TMAH-based Py-MS instrument. One of idea to solve the knotty problem has been flashed from "Top-down proteomics for identification of intact microoganisms". That is, biomarker peaks are selected from complicate Py-MS spectra for intact microoganisms by tracing out their origins, based on Py-MS spectra for the featured components of different cell-types, in Top-down approach. This idea has been tested in classification of different Gram-type microoganisms. Through the analyses of spectra for the featured components - peptidoglycan and lipoteichoic acid for Gram-positive cells and lipopolysaccharide and lipid A for Gram-negative cells - with comparing to the spectra the corresponding Gram-type cells in the Top-down approach, biomarker peaks were selected to carry out PCA(Principal Component Analysis) in order to see classification of different Gram-types, resulting in significant improvement of their classification. Furthermore, weighting biomarker peaks on intact cell's spectra, based on the data for the featured components of the Gram-types, contributed to elevate classification performance.

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