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Development and comparison of novel multiple cross displacement amplification (MCDA) assays with other nucleic acid amplification methods for SARS-CoV-2 detection.
Luu, Laurence Don Wai; Payne, Michael; Zhang, Xiaomei; Luo, Lijuan; Lan, Ruiting.
  • Luu LDW; School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia.
  • Payne M; School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia.
  • Zhang X; School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia.
  • Luo L; School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia.
  • Lan R; School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia. r.lan@unsw.edu.au.
Sci Rep ; 11(1): 1873, 2021 01 21.
Article in English | MEDLINE | ID: covidwho-1043028
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ABSTRACT
The development of alternative isothermal amplification assays including multiple cross displacement amplification (MCDA) may address speed and portability limitations of real-time PCR (rt-PCR) methods for SARS-CoV-2 detection. We developed a novel SARS-CoV-2 MCDA assay and compared its speed and sensitivity to loop-mediated isothermal amplification (LAMP) and rt-PCR. Two MCDA assays targeting SARS-CoV-2 N gene and ORF1ab were designed. The fastest time to detection and sensitivity of MCDA was compared to LAMP and rt-PCR using DNA standards and transcribed RNA. For the N gene, MCDA was faster than LAMP and rt-PCR by 10 and 20 min, respectively with fastest time to detection at 5.2 min. rt-PCR had the highest sensitivity with the limit of detection at 10 copies/µl compared with MCDA (100 copies/µl) and LAMP (500 copies/µl). For ORF1ab, MCDA and LAMP had similar speed with fastest time to detection at 9.7 and 8.4 min, respectively. LAMP was more sensitive for ORF1ab detection with 50 copies/µl compared to MCDA (500 copies/µl). In conclusion, different nucleic acid amplification methods provide different advantages. MCDA is the fastest nucleic acid amplification method for SARS-CoV-2 while rt-PCR is the most sensitive. These advantages should be considered when determining the most suitable nucleic acid amplification methods for different applications.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Nucleic Acid Amplification Techniques / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study / Randomized controlled trials Limits: Humans Language: English Journal: Sci Rep Year: 2021 Document Type: Article Affiliation country: S41598-021-81518-8

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Nucleic Acid Amplification Techniques / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study / Randomized controlled trials Limits: Humans Language: English Journal: Sci Rep Year: 2021 Document Type: Article Affiliation country: S41598-021-81518-8