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Updated: Oct 20, 2025
Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification
Published on: March 31, 2021
Diagnostic evaluation of qRT-PCR-based kit and dPCR-based kit for COVID-19
Cherl-Joon Lee1, Wonseok Shin2, Seyoung Mun3,4,5
1Department of Bio-Convergence Engineering, Dankook University, Jukjeon, 16890, Republic of Korea.
Insights
This study compared two COVID-19 diagnostic kits. The Dr. PCR 20K kit demonstrated superior sensitivity, detecting lower viral loads for accurate COVID-19 diagnosis.
Area of Science:
- Molecular Diagnostics
- Infectious Disease Detection
- Biotechnology
Background:
- COVID-19, a fatal respiratory illness, requires early diagnosis to curb global spread.
- Accurate and efficient diagnostic tools are essential for managing the pandemic.
Purpose of the Study:
- To evaluate the diagnostic performance of the Allplex™ 2019-nCoV Assay (qRT-PCR) and Dr. PCR 20K COVID-19 Detection kit (dPCR).
- To compare the sensitivity and efficiency of qRT-PCR and dPCR technologies for COVID-19 detection.
Main Methods:
- Utilized 30 negative and 20 positive COVID-19 specimens for diagnostic testing.
- Assessed diagnostic accuracy using sensitivity, specificity, and Cohen's Kappa coefficient.
- Conducted dilutional linearity experiments to compare detection limits.
Main Results:
- Both kits achieved identical diagnostic results for all specimens, with perfect agreement (Cohen's Kappa = 1).
- The Dr. PCR 20K kit exhibited a tenfold lower detection limit for SARS-CoV-2 viral load compared to the Allplex™ kit.
- Statistical analysis confirmed high diagnostic accuracy for both methods.
Conclusions:
- Digital PCR (dPCR) offers a powerful alternative for rapid COVID-19 point-of-care testing.
- dPCR's advantages include smaller equipment size and suitability for limited settings.
- The Dr. PCR 20K kit shows promise for efficient and sensitive COVID-19 diagnosis.
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.
