Diagnostic evaluation of qRT-PCR-based kit and dPCR-based kit for COVID-19

Abstract

Background Coronavirus disease of 2019 (COVID-19) is well known as a fatal disease, first discovered at Wuhan in China, ranging from mild to death, such as shortness of breath and fever. Early diagnosis of COVID-19 is a crucial point in preventing global prevalence. Objective We aimed to evaluate the diagnostic competency and efficiency with the Allplex™ 2019-nCoV Assay kit and the Dr. PCR 20 K COVID-19 Detection kit, designed based on the qRT-PCR and dPCR technologies, respectively. Methods A total of 30 negative and 20 COVID-19 positive specimens were assigned to the diagnostic test by using different COVID-19 diagnosis kits. Diagnostic accuracy was measured by statistical testing with sensitivity, specificity, and co-efficiency calculations. Results Comparing both diagnostic kits, we confirmed that the diagnostic results of 30 negative and 20 positive cases were the same pre-diagnostic results. The diagnostic statistics test results were perfectly matched with value (1). Cohen’s Kappa coefficient was demonstrated that the given kits in two different ways were “almost perfect” with value (1). In evaluating the detection capability, the dilutional linearity experiments substantiate that the Dr. PCR 20 K COVID-19 Detection kit could detect SARS-CoV-2 viral load at a concentration ten times lower than that of the Allplex™ 2019-nCoV Assay kit. Conclusions In this study, we propose that the dPCR diagnosis using LOAA dPCR could be a powerful method for COVID-19 point-of-care tests requiring immediate diagnosis in a limited time and space through the advantages of relatively low sample concentration and small equipment size compared to conventional qRT-PCR. Supplementary Information The online version contains supplementary material available at 10.1007/s13258-021-01162-4.

Diagnostic, Prognostic, and Therapeutic Value of Droplet Digital PCR (ddPCR) in COVID-19 Patients: A Systematic Review

“…The studies were conducted worldwide; 16 in Asia [ 12 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 ], 14 in Europe [ 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 ], and 9 in North America [ 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 ]. A minority of studies (8) included control samples [ 21 , 28 , 29 , 33 , 38 , 47 , 54 , 56 ]. One study did not report the number of samples or patients studied [ 26 ].…”

Rapid identification of SARS-CoV-2 in the point-of-care using digital PCR-based Dr. PCR™ Di20K COVID-19 Detection Kit without viral RNA extraction

“…In a previous study, LOAA dPCR equipment (Optolane, Seongnam, Republic of Korea) with dPCR-based Dr. PCR 20 K COVID-19 Detection Kit (Cat no. DCM402-X, Optolane, Seongnam, Republic of Korea) system has been reported that it is less affected by PCR inhibitors than the typical qRT-PCR assay and can be detected even at low viral loads (Lee et al 2021a ; Sidstedt et al 2020 ). In addition, this system has high diagnostic sensitivity and specificity.…”

Update on the Development of Toehold Switch-Based Approach for Molecular Diagnostic Tests of COVID-19

“…These fragments can detect unique viral RNA sequences in nucleocapsid (N), envelope (E), spike (S), or RNA-dependent RNA polymerase (RdRp) genes using the fluorescence signals [ 13 , 14 ]. The mentioned method is currently considered the “gold standard” in COVID-19 diagnostics due to its high sensitivity and specificity and its straightforward quantitative analysis [ 15 , 16 ]. However, SARS-CoV-2 diagnostic using RT-qPCR requires substantial hours and relatively expensive instruments, chemicals, and consumables to generate results [ 17 ].…”