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RNA purification-free detection of SARS-CoV-2 using reverse transcription loop-mediated isothermal amplification (RT-LAMP)

Lai MY 1 , Suppiah J 2 , Thayan R 2 , Ismail I 3 , Mustapa NI 3 , Soh TST 3 Show all authors , Hassan AH 3 , Peariasamy KM 4 , Lee YL 5 , Lau YL 6

Affiliations 

  • 1 Department of Parasitology, Faculty of Medicine, University Malaya, 50603, Kuala Lumpur, Malaysia
  • 2 Virology Unit, Infectious Disease Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health, Shah Alam, Selangor, Malaysia
  • 3 Department of Pathology, Hospital Sungai Buloh, Ministry of Health, Sungai Buloh, Selangor, Malaysia
  • 4 Institute for Clinical Research, National Institutes of Health, Ministry of Health, Shah Alam, Selangor, Malaysia
  • 5 Clinical Research Centre, Hospital Sungai Buloh, Ministry of Health, Sungai Buloh, Selangor, Malaysia
  • 6 Department of Parasitology, Faculty of Medicine, University Malaya, 50603, Kuala Lumpur, Malaysia. [email protected]
Trop Med Health, 2022 Jan 04;50(1):2.
PMID: 34980275 DOI: 10.1186/s41182-021-00396-y

Abstract

BACKGROUND: Current diagnosis of SARS-CoV-2 infection relies on RNA purification prior to amplification. Typical extraction methods limit the processing speed and turnaround time for SARS-CoV-2 diagnostic testing.

METHODS: Here, we applied reverse transcription loop-mediated isothermal amplification directly onto human clinical swabs samples to amplify the RNA from SARS-CoV-2 swab samples after processing with chelating resin.

RESULTS: By testing our method on 64 samples, we managed to develop an RT-LAMP assay with 95.9% sensitivity (95% CI 86 to 99.5%) and 100% specificity (95% CI 78.2-100%).

CONCLUSION: The entire process including sample processing can be completed in approximately 50 min. This method has promising potential to be applied as a fast, simple and inexpensive diagnostic tool for the detection of SARS-CoV-2.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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