Combination of RT-LAMP and fluorescence spectroscopy using chemometric techniques for an ultra-sensitive and rapid alternative for the detection of SARS-CoV-2

Abstract

The increased spread of COVID-19 caused by SARS-CoV-2 has made it necessary to develop more efficient, fast, accurate, specific, sensitive and easy-to-use detection platforms to overcome the disadvantages of gold standard methods (RT-qPCR). Here an approach was developed for the detection of the SARS-CoV-2 virus using the loop-mediated isothermal amplification (LAMP) technique for SARS-CoV-2 RNA target amplification in samples of nasopharyngeal swabs. The discrimination between positive and negative SARS-CoV-2 samples was achieved by using fluorescence spectra generated by the excitation of the LAMP's DNA intercalator dye at λ497 nm in a fluorescence spectrophotometer and chemometric tools. Exploratory analysis of the 83 sample spectra using principal component analysis (PCA) indicated a trend in differentiation between positive and negative samples resulting from the peak emission of the fluorescent dye. The classification was performed by partial least squares discriminant analysis (PLS-DA) achieving a sensitivity, a specificity and an accuracy of 100%, 95% and 89%, respectively for the discrimination between negative and positive samples from 1.58 to 0.25 ng L⁻¹ after LAMP amplification. Therefore, this study indicates that the use of the LAMP technique in fluorescence spectroscopy may offer a fast (<1 hour), sensitive and low-cost method.