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Clinical testing on SARS-CoV-2 swab samples using reverse-transcription loop-mediated isothermal amplification
Meng Yee Lai1, Fatma Diyana Mohd Bukhari1, Nur Zulaikha Zulkefli1
1Department of Parasitology, Faculty of Medicine, Universiti Malaya, 50603, Kuala Lumpur, Malaysia.
BMC Infectious Diseases
|August 18, 2022
Summary
A new, simple nucleic acid extraction method for SARS-CoV-2 detection offers a cost-effective alternative to commercial kits. This method, combined with RT-LAMP, shows high sensitivity and specificity for point-of-care diagnostics.
Area of Science:
- Molecular Biology
- Virology
- Diagnostic Development
Background:
- High cost of commercial RNA extraction kits hinders efficient SARS-CoV-2 testing.
- Development of a simplified nucleic acid extraction method is crucial for widespread diagnostics.
Purpose of the Study:
- To develop and evaluate a simple nucleic acid extraction method for SARS-CoV-2 detection.
- To assess the feasibility of this method for point-of-care diagnostics.
Main Methods:
- A novel, simple nucleic acid extraction technique was developed for direct use with nasopharyngeal swab samples.
- Reverse transcription loop-mediated isothermal amplification (RT-LAMP) was employed for viral detection.
- A pH-sensitive dye was utilized for endpoint detection, indicated by visible color changes.
Main Results:
- Clinical evaluation involved 260 samples.
- The RT-LAMP assay demonstrated 92.7% sensitivity (95% CI 87.3-96.3%) and 93.6% specificity (95% CI 87.3-97.4%).
- The colorimetric endpoint detection eliminated the need for specialized laboratory equipment.
Conclusions:
- The developed nucleic acid extraction method simplifies the process and reduces the need for extensive laboratory infrastructure.
- Combining this method with RT-LAMP technology presents a promising point-of-care diagnostic tool for SARS-CoV-2.
