[논문]Comparison of Digital PCR and Quantitative PCR with Various SARS-CoV-2 Primer-Probe Sets

Park, Changwoo; Lee, Jina; Hassan, Zohaib ul; Ku, Keun Bon; Kim, Seong-Jun; Kim, Hong Gi; Park, Edmond Changkyun; Park, Gun-Soo; Park, Daeui; Baek, Seung-Hwa; Park, Dongju; Lee, Jihye; Jeon, Sangeun; Kim, Seungtaek; Lee, Chang-Seop; Yoo, Hee Min; Kim, Seil

doi:10.4014/jmb.2009.09006

[국내논문] Comparison of Digital PCR and Quantitative PCR with Various SARS-CoV-2 Primer-Probe Sets 원문보기

Journal of microbiology and biotechnology, v.31 no.3, 2021년, pp.358 - 367

Park, Changwoo (Microbiological Analysis Team, Biometrology Group, Korea Research Institute of Standards and Science (KRISS)) , Lee, Jina (Microbiological Analysis Team, Biometrology Group, Korea Research Institute of Standards and Science (KRISS)) , Hassan, Zohaib ul (Microbiological Analysis Team, Biometrology Group, Korea Research Institute of Standards and Science (KRISS)) , Ku, Keun Bon (Center for Convergent Research of Emerging Virus Infection, Korea Research Institute of Chemical Technology) , Kim, Seong-Jun (Center for Convergent Research of Emerging Virus Infection, Korea Research Institute of Chemical Technology) , Kim, Hong Gi (Center for Convergent Research of Emerging Virus Infection, Korea Research Institute of Chemical Technology) , Park, Edmond Changkyun (Center for Convergent Research of Emerging Virus Infection, Korea Research Institute of Chemical Technology) , Park, Gun-Soo (Center for Convergent Research of Emerging Virus Infection, Korea Research Institute of Chemical Technology) , Park, Daeui (Center for Convergent Research of Emerging Virus Infection, Korea Research Institute of Chemical Technology) , Baek, Seung-Hwa (Center) , Park, Dongju , Lee, Jihye , Jeon, Sangeun , Kim, Seungtaek , Lee, Chang-Seop , Yoo, Hee Min , Kim, Seil

Abstract ▼ AI-Helper

The World Health Organization (WHO) has declared the coronavirus disease 2019 (COVID-19) as an international health emergency. Current diagnostic tests are based on the reverse transcription-quantitative polymerase chain reaction (RT-qPCR) method, which is the gold standard test that involves the amplification of viral RNA. However, the RT-qPCR assay has limitations in terms of sensitivity and quantification. In this study, we tested both qPCR and droplet digital PCR (ddPCR) to detect low amounts of viral RNA. The cycle threshold (CT) of the viral RNA by RT-PCR significantly varied according to the sequences of the primer and probe sets with in vitro transcript (IVT) RNA or viral RNA as templates, whereas the copy number of the viral RNA by ddPCR was effectively quantified with IVT RNA, cultured viral RNA, and RNA from clinical samples. Furthermore, the clinical samples were assayed via both methods, and the sensitivity of the ddPCR was determined to be equal to or more than that of the RT-qPCR. However, the ddPCR assay is more suitable for determining the copy number of reference materials. These findings suggest that the qPCR assay with the ddPCR defined reference materials could be used as a highly sensitive and compatible diagnostic method for viral RNA detection.

Keyword

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용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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