A colorimetric reverse transcription-loop mediated isothermal amplification (RT-LAMP) method for the rapid detection of SARS-CoV-2 in Thailand
https://doi.org/10.47665/tb.41.1.008
- Author:
Bhakdeenuan, P.
1
;
Bunchoo, S.
1
;
Klayut, K.
1
;
Srisungngam, S.
1
;
Suphan, O.
2
;
Kongthap, I.
2
;
Suphankong, S.
3
;
Phetsuksiri, B.
4
;
Uppapong, B.
1
;
Rudeeaneksin, J.
1
Author Information
1. National Institute of Health, Department of Medical Sciences, Ministry of Public Health, Nonthaburi, Thailand&
2. Regional Medical Sciences Center 11/1 Phuket, Department of Medical Sciences, Ministry of Public Health, Nonthaburi, Thailand
3. Regional Medical Sciences Center 5 Samut Songkhram, Department of Medical Sciences, Ministry of Public Health, Nonthaburi, Thailand
4. National Institute of Health, Department of Medical Sciences, Ministry of Public Health, Nonthaburi, Thailand&Medical Sciences Technical Office, Department of Medical Sciences, Ministry of Public Health, Nonthaburi, Thailand
- Publication Type:Journal Article
- Keywords:
SARS-CoV-2;
COVID-19;
RT-LAMP;
RT-PCR;
validation.
- From:Tropical Biomedicine
2024;41(No.1):64-69
- CountryMalaysia
- Language:English
-
Abstract:
COVID-19, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), remains a global
health threat. Timely identification of infected cases is important for appropriate patient management
and the control of viral spread. Simple and cost-effective tests are required to increase access to
testing and early case detection. Here, we describe a colorimetric reverse transcription-loop-mediated
isothermal amplification (RT-LAMP) method to detect SARS-CoV-2. The RT-LAMP could amplify the
orf1ab sequence detectable by visual color change within 45 min at 63 °C. The limit of detection (LoD)
for SARS-CoV-2 RNA was less than 100 copies (13.36) per reaction with no cross-amplification with other
related viruses. Clinical evaluation using leftover RNA samples extracted from 163 nasopharyngeal swab
specimens showed perfect agreement in negative (n = 124) and positive samples with cycle thresholds
(Ct) < 34 cycles (n = 33) detected by real-time reverse transcription-polymerase chain reaction (RT-PCR),
targeting RdRp and N genes as a reference. Overall, the diagnostic accuracy, sensitivity, specificity,
positive and negative predictive values of RT-LAMP in testing were 96.32% (95% CI: 92.16-98.64%),
84.62% (95% CI: 68.47-94.14%), 100% (95% CI: 97.07-100.0%), 100% (95% CI: 89.42-100.0%), and 95.38%
(95% CI: 90.22-98.29), respectively. This RT-LAMP assay is simple and reliable, with the potential to be
an alternative for the rapid detection of SAR-CoV-2 with minimal time and fewer resources compared
to real-time RT-PCR.
- Full text:20240711121841431628.2024my1454.pdf
