The Analytical Application of Fluorescence Spectrophotometry for Identifying SARS-CoV-2

Madurani, Kartika Anoraga (2021) The Analytical Application of Fluorescence Spectrophotometry for Identifying SARS-CoV-2. Doctoral thesis, Institut Teknologi Sepuluh Nopember.

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Fluorescence spectrophotometry has been applied to identify Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). SARS-CoV-2 is a new coronavirus that causes Coronavirus Disease 2019 or COVID-19. Clinical swab samples from positive and negative COVID-19 were collected from 80 patients using a sterile cotton swab. The samples were obtained from the nasopharyngeal and oropharyngeal swabs of the patients. These sterile cotton swabs containing the samples were kept in RADI transport medium (RTM) prior to use. The samples in RTM are divided into two groups. The first group of the samples was analyzed using real-time reverse transcription-polymerase chain reaction (RT-qPCR) to identify the positive and negative COVID-19 as well as to be the control. The second group of the samples was analyzed using fluorescence spectrophotometry analysis. Fluorescence measurement was carried out at excitation and emission wavelength from 200-800 nm. Forty positives and four negatives COVID-19 samples were available from the result of RT-qPCR. Twenty positives and twenty negatives COVID-19 were taken 0.5 mL, and the other twenty positives and twenty negatives COVID-19 were taken 1 mL for fluorescence measurement. The best results were obtained at volume samples of 1 mL. The obtained data of fluorescence spectrum showed that the samples with and without COVID-19 infection have significantly different characteristics. These can be observed in emission peak wavelength at 522.47±11.78 nm and 692.18±10.89 nm for positive COVID-19 and 508.37±5.68 nm 685.27±0.44 nm for negative COVID-19. All positive samples can distinguish from the negative sample using fluorescence spectrophotometry. Principle Component Analysis (PCA) showed that positive COVID-19 samples have a different location from negative COVID-19 samples. The effect of various Ct values from positive samples were not shown different fluorescence characteristics. 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 takes 4 min. This technique offers an efficient and accurate method to identify an active infected individual and be easily adapted to early COVID-19 investigation in general.

Item Type: Thesis (Doctoral)
Uncontrolled Keywords: SARS-CoV-2 infection; fast analysis; virus; fluorescence spectroscopy; excitation; emission.
Subjects: Q Science > QD Chemistry > QD251.2 Chemistry, Organic. Biochemistry
Q Science > QD Chemistry > QD75.2 Chemistry, Analytic
Q Science > QD Chemistry > QD96F56 Fluorescence spectroscopy
Divisions: Faculty of Science and Data Analytics (SCIENTICS) > Chemistry > 47001-(S3) PhD Thesis
Depositing User: Kartika Anoraga Madurani
Date Deposited: 24 Aug 2021 07:12
Last Modified: 24 Aug 2021 07:12

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