Author: Luu, Laurence Don Wai; Payne, Michael J; Zhang, Xiaomei; Luo, Lijuan; Lan, Ruiting
ID: sazv7iow
Snippet: 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 was designed. The fastest time to detection and sensitivity of MCDA was co
Document: 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 was designed. The fastest time to detection and sensitivity of MCDA was compared to LAMP and rt-PCR using DNA standards and transcribed RNA. For N gene, MCDA was faster than LAMP and rt-PCR by 10 and 20 minutes, respectively with fastest time to detection at 5.2 minutes. rt-PCR had highest sensitivity with limit of detection at 10 copies/{micro}l compared with MCDA (100 copies/{micro}l) and LAMP (500 copies/{micro}l). For ORF1ab, MCDA and LAMP had similar speed with fastest time to detection at 9.7 and 8.4 minutes, respectively. LAMP was more sensitive for ORF1ab detection with 50 copies/{micro}l compared to MCDA (500 copies/{micro}l). 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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