Optimization of extraction-free protocols for SARS-CoV-2 detection using a commercial rRT-PCR assayopen access
- Authors
- Kang, Minhee; Jeong, Eunjung; Kim, Ji-Yeon; Yun, Sun Ae; Jang, Mi-Ae; Jang, Ja-Hyun; Kim, Tae Yeul; Huh, Hee Jae; Lee, Nam Yong
- Issue Date
- Dec-2023
- Publisher
- Nature Research
- Citation
- Scientific Reports, v.13, no.1
- Indexed
- SCIE
SCOPUS
- Journal Title
- Scientific Reports
- Volume
- 13
- Number
- 1
- URI
- https://scholarworks.bwise.kr/skku/handle/2021.sw.skku/109561
- DOI
- 10.1038/s41598-023-47645-0
- ISSN
- 2045-2322
- Abstract
- In the ongoing global fight against coronavirus disease 2019 (COVID-19), the sample preparation process for real-time reverse transcription polymerase chain reaction (rRT-PCR) faces challenges due to time-consuming steps, labor-intensive procedures, contamination risks, resource demands, and environmental implications. However, optimized strategies for sample preparation have been poorly investigated, and the combination of RNase inhibitors and Proteinase K has been rarely considered. Hence, we investigated combinations of several extraction-free protocols incorporating heat treatment, sample dilution, and Proteinase K and RNase inhibitors, and validated the effectiveness using 120 SARS-CoV-2 positive and 62 negative clinical samples. Combining sample dilution and heat treatment with Proteinase K and RNase inhibitors addition exhibited the highest sensitivity (84.26%) with a mean increase in cycle threshold (Ct) value of + 3.8. Meanwhile, combined sample dilution and heat treatment exhibited a sensitivity of 79.63%, accounting for a 38% increase compared to heat treatment alone. Our findings highlight that the incorporation of Proteinase K and RNase inhibitors with sample dilution and heat treatment contributed only marginally to the improvement without yielding statistically significant differences. Sample dilution significantly impacts SARS-CoV-2 detection, and sample conditions play a crucial role in the efficiency of extraction-free methods. Our findings may provide insights for streamlining diagnostic testing, enhancing its accessibility, cost-effectiveness, and sustainability. © 2023, The Author(s).
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Collections - Medicine > Department of Medicine > 1. Journal Articles
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