Dual-target one-step nested PCR for sensitive detection of SARS-CoV-2 nucleic acids

Qijie Li, Yiqing Xia, Dunshui Liao, Hu Nie, Ming Zhang, Tinghua Wang, Jiayu Liao, Qingjie Xia*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

The Corona Virus Disease reported in 2019 (COVID-19) poses a significant threat to human and public health. Its early and accurate detection can reduce the spread and recurrence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Real-time reverse transcription fluorescent quantitative polymerase chain reaction (RT-qPCR) is the “gold standard” for detecting the nucleic acid of SARS-CoV-2. This study developed and tested a dual-target (ORF1ab and N gene) one-step nested RT-qPCR (DTO-N-PCR) to detect SARS-CoV-2. Ten-fold serial dilutions of mixed synthetic DNA from SARS-CoV-2 ORF1ab and N gene were used as templates to test the sensitivity of DTO-N-PCR. Its specificity was subsequently tested using throat swab specimens from 10 COVID-19 patients and 35 healthy participants. DTO-N-PCR was more sensitive and specific than conventional RT-qPCR. It has unique features, including a dual-target (ORF1ab and N gene), rapid one-step operation of reverse transcription and PCR, four pairs of inner and outer primers, and specific probes. These features aid in its rapid, accurate, and efficient detection of SARS-CoV-2 RNA.

Original languageEnglish
Pages (from-to)471-477
Number of pages7
JournalPreparative Biochemistry and Biotechnology
Volume52
Issue number4
DOIs
Publication statusPublished - 1 Apr 2022
Externally publishedYes

Keywords

  • COVID-19
  • detection
  • nucleic acid
  • RT-qPCR
  • SARS-CoV-2

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