Two extraction-free reverse transcription loop-mediated isothermal amplification assays for detection of SARS-CoV-2

Author:

Lai Meng Yee,Bukhari Fatma Diyana Mohd,Zulkefli Nur Zulaikha,Ismail Ilyiana,Mustapa Nur Izati,Soh Tuan Suhaila Tuan,Hassan Afifah Haji,Peariasamy Kalaiarasu M.,Lee Yee Leng,Suppiah Jeyanthi,Thayan Ravindran,Lau Yee Ling

Abstract

Abstract Background Current assays for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rely on time consuming, costly and laboratory based methods for virus isolation, purification and removing inhibitors. To address this limitation, we propose a simple method for testing RNA from nasopharyngeal swab samples that bypasses the RNA purification step. Methods In the current project, we have described two extraction-free reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays for the detection of SARS-CoV-2 by using E gene and RdRp gene as the targets. Results Here, results showed that reverse transcription loop-mediated isothermal amplification assays with 88.4% sensitive (95% CI: 74.9–96.1%) and 67.4% sensitive (95% CI: 51.5–80.9%) for E gene and RdRp gene, respectively. Conclusion Without the need of RNA purification, our developed RT-LAMP assays for direct detection of SARS-CoV-2 from nasopharyngeal swab samples could be turned into alternatives to qRT-PCR for rapid screening.

Funder

Prototype Research Grant Scheme (PRGS), Ministry of Education, Malaysia

Publisher

Springer Science and Business Media LLC

Subject

Infectious Diseases

Reference24 articles.

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