Detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in clinical samples using Real-Time Reverse Transcription Polymerase Chain Reaction (qRT-PCR)

Abstract

The coronavirus disease 2019 (COVID-19) is caused by an infection with the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), which has infected more than 400,000 people, causing a worldwide pandemic. As of August 15, 2020, as many as 111,455 cases of COVID-19 have been confirmed in Indonesia with 6,071 deaths that have been recorded thus far. Quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) has been used as a gold standard to detect the infection of SARS-CoV-2 in many clinical samples due to its sensitivity and specificity. In Indonesia, one of the recommended and commonly used diagnostic kit for COVID-19 was MiRXES Fortitude Kit 2.0E under the license of Singapore's Agency for Science, Technology, and Research (A*STAR), Singapore. Aiming to assess the basic analytical and clinical performance of the MiRXES Fortitude Kit 2.0E, this study prepares serial dilutions of the provided positive control to establish PCR efficiency and to estimate the limit of detection. Moreover, a panel of SARS-CoV-2-positive-clinical samples (nasopharyngeal swab specimens) was tested to preliminarily evaluate the kit's sensitivity on clinical samples. This study is expected to provide valuable assessment information for laboratories that develop routine diagnostic protocols for COVID-19, especially those using the MiRXES Fortitude Kit (A*Star, Singapore).