Ultrarapid On-Site Detection of SARS-CoV-2 Infection Using Simple ATR-FTIR Spectroscopy and an Analysis Algorithm: High Sensitivity and Specificity.

Affiliation

Barauna VG(1), Singh MN(2), Barbosa LL(1), Marcarini WD(1), Vassallo PF(1)(3), Mill JG(1), Ribeiro-Rodrigues R(4), Campos LCG(5), Warnke PH(6)(7), Martin FL(2).
Author information:
(1)Department of Physiological Sciences, Federal University of Espírito Santo, 29075-910 Vitoria, Brazil.
(2)Biocel UK Ltd., 15 Riplingham Road, West Ella, Hull HU10 6TS, U.K.
(3)Clinical Hospital, Federal University of Minas Gerais, 31270-901 Belo Horizonte, Brazil.
(4)Núcleo de Doenças Infecciosas, Federal University of Espírito Santo, 29075-910 Vitoria, Brazil.
(5)Department of Biological Science, Santa Cruz State University, 45662-900 Bahia, Brazil.
(6)Praxisklinik am Ballastkai, Ballastkai 5, 24937 Flensburg, Germany.
(7)Department of OMF-Surgery, Christian-Albrechts-University of Kiel, 24118 Kiel, Germany.

Abstract

There is an urgent need for ultrarapid testing regimens to detect the severe acute respiratory syndrome coronavirus 2 [SARS-CoV-2] infections in real-time within seconds to stop its spread. Current testing approaches for this RNA virus focus primarily on diagnosis by RT-qPCR, which is time-consuming, costly, often inaccurate, and impractical for general population rollout due to the need for laboratory processing. The latency until the test result arrives with the patient has led to further virus spread. Furthermore, latest antigen rapid tests still require 15-30 min processing time and are challenging to handle. Despite increased polymerase chain reaction (PCR)-test and antigen-test efforts, the pandemic continues to evolve worldwide. Herein, we developed a superfast, reagent-free, and nondestructive approach of attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy with subsequent chemometric analysis toward the prescreening of virus-infected samples. Contrived saliva samples spiked with inactivated γ-irradiated COVID-19 virus particles at levels down to 1582 copies/mL generated infrared (IR) spectra with a good signal-to-noise ratio. Predominant virus spectral peaks are tentatively associated with nucleic acid bands, including RNA. At low copy numbers, the presence of a virus particle was found to be capable of modifying the IR spectral signature of saliva, again with discriminating wavenumbers primarily associated with RNA. Discrimination was also achievable following ATR-FTIR spectral analysis of swabs immersed in saliva variously spiked with virus. Next, we nested our test system in a clinical setting wherein participants were recruited to provide demographic details, symptoms, parallel RT-qPCR testing, and the acquisition of pharyngeal swabs for ATR-FTIR spectral analysis. Initial categorization of swab samples into negative versus positive COVID-19 infection was based on symptoms and PCR results (n = 111 negatives and 70 positives). Following training and validation (using n = 61 negatives and 20 positives) of a genetic algorithm-linear discriminant analysis (GA-LDA) algorithm, a blind sensitivity of 95% and specificity of 89% was achieved. This prompt approach generates results within 2 min and is applicable in areas with increased people traffic that require sudden test results such as airports, events, or gate controls.