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Evaluation of diagnostic performance of SARS-CoV-2 infection using digital droplet polymerase chain reaction in individuals with or without COVID-19 symptoms

Authors
 Yoonjung Kim  ;  Eunyoung Lee  ;  Boyeon Kim  ;  Jinhee Cho  ;  Sook-Won Ryu  ;  Kyung-A Lee 
Citation
 CLINICA CHIMICA ACTA, Vol.554 : 117759, 2024-02 
Journal Title
CLINICA CHIMICA ACTA
ISSN
 0009-8981 
Issue Date
2024-02
MeSH
COVID-19 Testing ; COVID-19* / diagnosis ; Humans ; Limit of Detection ; RNA, Viral / genetics ; Real-Time Polymerase Chain Reaction / methods ; SARS-CoV-2 / genetics ; Sensitivity and Specificity ; Viral Load / methods
Keywords
Digital droplet polymerase chain reaction ; RT-qPCR ; SARS-CoV-2
Abstract
Background: Reverse transcription-quantitative PCR (RT-qPCR) is commonly used to diagnose SARS-CoV-2, but it has limited sensitivity in detecting the virus in asymptomatic close contacts and convalescent patients. In this study, we propose the use of reverse transcription-digital droplet PCR (RT-ddPCR) to detect SARS-CoV-2 in clinical samples. Methods: The clinical performance of RT-ddPCR targeting of ORF1ab and N genes was evaluated in parallel with RT-qPCR using 200 respiratory samples collected from close contacts and patients at different phases of infection. Results: The limits of detection (LODs) for RT-ddPCR assays were determined using six dilutions of ACCUPLEX SARS-Cov-2 reference material. The LODs of ORF1ab and N genes were 3.7 copies/reaction and 2.2 copies/reaction, respectively. Compared to RT-qPCR, RT-ddPCR increased the positive rate by 12.0% in 142 samples from SARS-CoV-2-infected patients. Additionally, RT-ddPCR detected SARS-CoV-2 in three of 26 specimens from close contacts that tested negative by RT-qPCR, and infection was confirmed using follow-up samples. Finally, RT-ddPCR improved the equivocal results from RT-qPCR in 56.3% (9/16) of convalescent patient samples. Conclusions: Detecting SARS-CoV-2 in samples with low viral loads using RT-qPCR can be challenging. However, our study suggests that RT-ddPCR, with its higher sensitivity and accuracy, is better suited for detecting low viral copies in samples, particularly those from close contacts and convalescent patients. © 2024 Elsevier B.V.
Full Text
https://www.sciencedirect.com/science/article/pii/S0009898123005612
DOI
10.1016/j.cca.2023.117759
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Laboratory Medicine (진단검사의학교실) > 1. Journal Papers
Yonsei Authors
Kim, Boyeon(김보연)
Kim, Yoon Jung(김윤정) ORCID logo https://orcid.org/0000-0002-4370-4265
Lee, Kyung A(이경아) ORCID logo https://orcid.org/0000-0001-5320-6705
Lee, Eun Young(이은영)
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/198608
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