Madurani Kartika A, Yudha Syahputra Muhammad, Puspita Ika, Furqoni Abdul Hadi, Puspasari Listya, Rosyidah Hafildatur, Hatta Agus Muhamad, Lusida Maria Inge, Tominaga Masato, Kurniawan Fredy
Laboratory of Instrumentation and Analytical Science, Chemistry Department, Faculty of Science and Data Analytics, Institut Teknologi Sepuluh Nopember, Surabaya 60111, Indonesia.
Photonics Engineering Laboratory, Department of Engineering Physics, Faculty of Industrial Technology and Systems Engineering, Institut Teknologi Sepuluh Nopember, Surabaya 60111, Indonesia.
Arab J Chem. 2022 Aug;15(8):104020. doi: 10.1016/j.arabjc.2022.104020. Epub 2022 May 30.
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.
考虑到目前用于检测严重急性呼吸综合征冠状病毒2（SARS-CoV-2）及其新出现变体的检测方法存在局限性，开发了一种使用荧光分光光度法的简单快速方法来检测2019冠状病毒病（COVID-19）。从COVID-19阳性和阴性患者的鼻咽和口咽腔收集了40份临床拭子样本。每个样本分为两部分。第一部分样本采用逆转录聚合酶链反应（RT-qPCR）作为对照方法进行分析，以鉴定COVID-19阳性和阴性样本。第二部分样本采用荧光分光光度法进行分析。在200至800nm的激发和发射波长下进行荧光测量。根据RT-qPCR结果检测了20份COVID-19阳性样本和20份COVID-19阴性样本。荧光光谱数据表明，COVID-19阳性和阴性样本具有明显不同的特征。通过荧光分光光度法可以将所有阳性样本与阴性样本区分开来。主成分分析表明，COVID-19阳性样本与COVID-19阴性样本分别聚类。本实验的特异性和准确性均达到100%。E基因的检测限（LOD）为42.20拷贝/ml（Ct值为33.65个循环），ORF1ab基因的检测限为63.60拷贝/ml（Ct值为31.36个循环）。该鉴定过程仅需4分钟。因此，该技术提供了一种高效、准确的方法来识别患有活动性SARS-CoV-2感染的个体，总体上可轻松适用于COVID-19的早期调查。
