Spectrofluorimetric in tandem with principal component analysis for characterizing mycobacterium tuberculosis disease

Abstract

In this study, fluorescence spectroscopy (FS) in tandem with the chemometric technique has effectively been used for analyzing respiratory infectious disease Mycobacterium tuberculosis. This research work is based on the fluorescence emission of bacterium grown in acid-fast bacillus (AFB) culture. Samples were collected from TB suspected patients of various areas of Khyber Pakhtunkhwa, and cultured in the Provisional TB Reference Lab, Hayatabad Medical Complex Peshawar, Pakistan. The fluorescence spectra from all these samples have been acquired using the 405 nm wavelength as excitation. The differences in the fluorescence emission spectra between TB positive, TB negative, and cultured media samples has been found at peak emission wavelengths arising at 467 nm and 490 nm. The differences between TB positive and negative samples were seen both in the intensities as well as shift in their fluorescence emission peaks. For highlighting the spectral differences, principal component analysis (PCA) is applied on the fluorescence emission data collected from the three sets of samples. PCA clearly cluster the data into three different groups based on features extracted from their fluorescence emission data. These results are novel and suggest that FS can be used as alternate confirmation tool of mycobacterium through AFB culture on a mass scale. Furthermore, this technique is fast, cost effective and user friendly that could be used effectively for the early detection of mycobacterium bacillus in the suspected individual.