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. 2022 Aug;15(8):104020.
doi: 10.1016/j.arabjc.2022.104020. Epub 2022 May 30.

Fluorescence spectrophotometry for COVID-19 determination in clinical swab samples

Kartika A Madurani  1 , Suprapto  1 , Muhammad Yudha Syahputra  1 , Ika Puspita  2 , Abdul Hadi Furqoni  3 , Listya Puspasari  1 , Hafildatur Rosyidah  1 , Agus Muhamad Hatta  2 , Juniastuti  4   5 , Maria Inge Lusida  4   5 , Masato Tominaga  6 , Fredy Kurniawan  1
Affiliations

Fluorescence spectrophotometry for COVID-19 determination in clinical swab samples

Kartika A Madurani et al. Arab J Chem. 2022 Aug.

Abstract

Considering the limitations of the assays currently available for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its emerging variants, a simple and rapid method using fluorescence spectrophotometry was developed to detect coronavirus disease 2019 (COVID-19). Forty clinical swab samples were collected from the nasopharyngeal and oropharyngeal cavities of COVID-19-positive and -negative. Each sample was divided into two parts. The first part of the samples was analyzed using reverse transcription-polymerase chain reaction (RT-qPCR) as the control method to identify COVID-19-positive and -negative samples. The second part of the samples was analyzed using fluorescence spectrophotometry. Fluorescence measurements were performed at excitation and emission wavelengths ranging from 200 to 800 nm. Twenty COVID-19-positive samples and twenty COVID-19-negative samples were detected based on RT-qPCR results. The fluorescence spectrum data indicated that the COVID-19-positive and -negative samples had significantly different characteristics. All positive samples could be distinguished from negative samples by fluorescence spectrophotometry. Principal component analysis showed that COVID-19-positive samples were clustered separately from COVID-19-negative samples. The specificity and accuracy of this experiment reached 100%. Limit of detection (LOD) obtained 42.20 copies/ml (Ct value of 33.65 cycles) for E gene and 63.60 copies/ml (Ct value of 31.36 cycles) for ORF1ab gene. This identification process only required 4 min. Thus, this technique offers an efficient and accurate method to identify an individual with active SARS-CoV-2 infection and can be easily adapted for the early investigation of COVID-19, in general.

Keywords: COVID-19; Emission; Excitation; Fast analysis; Fluorescence spectroscopy; SARS-CoV-2.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Fig. 1
Fig. 1
Schematic representation of the fluorescence analysis process for detecting COVID-19-negative and -positive samples.
Fig. 2
Fig. 2
Excitation (A) and emission (B) peaks of negative (black) and positive (red) samples. The specific emission wavelength region: the first (yellow area) and the second (blue area).
Fig. 3
Fig. 3
Principal component analysis of COVID-19 samples.
See this image and copyright information in PMC

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