Lau, Yee Ling and Ismail, Ilyiana Binti and Mustapa, Nur Izati Binti and Lai, Meng Yee and Soh, Tuan Suhaila Tuan and Hassan, Afifah Haji and Peariasamy, Kalaiarasu M. and Lee, Yee Leng and Abdul Kahar, Maria Kahar Bador and Chong, Jennifer and Goh, Pik Pin (2021) Development of a reverse transcription recombinase polymerase amplification assay for rapid and direct visual detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). PLoS ONE, 16 (1). ISSN 1932-6203, DOI https://doi.org/10.1371/journal.pone.0245164.
Full text not available from this repository.Abstract
Rapid diagnosis is an important intervention in managing the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) outbreak. Real time reverse transcription polymerase chain reaction (RT-qPCR) remains the primary means for diagnosing the new virus strain but it is time consuming and costly. Recombinase polymerase amplification (RPA) is an isothermal amplification assay that does not require a PCR machine. It is an affordable, rapid, and simple assay. In this study, we developed and optimized a sensitive reverse transcription (RT)-RPA assay for the rapid detection of SARS-CoV-2 using SYBR Green I and/or lateral flow (LF) strip. The analytical sensitivity and specificity of the RT-RPA assay were tested by using 10-fold serial diluted synthetic RNA and genomic RNA of similar viruses, respectively. Clinical sensitivity and specificity of the RT-RPA assay were carried out using 78 positive and 35 negative nasopharyngeal samples. The detection limit of both RPA and RT-qPCR assays was 7.659 and 5 copies/mu L RNA, respectively with no cross reactivity with other viruses. The clinical sensitivity and specificity of RT-RPA were 98% and 100%, respectively. Our study showed that RT-RPA represents a viable alternative to RT-qPCR for the detection of SARS-CoV-2, especially in areas with limited infrastructure.
| Item Type: | Article |
|---|---|
| Funders: | UNSPECIFIED |
| Uncontrolled Keywords: | Uracil-DNA-glycosylase; Contamination |
| Subjects: | Q Science > QR Microbiology R Medicine > R Medicine (General) |
| Divisions: | Faculty of Medicine |
| Depositing User: | Ms Zaharah Ramly |
| Date Deposited: | 09 Jun 2022 07:29 |
| Last Modified: | 09 Jun 2022 07:29 |
| URI: | http://eprints.um.edu.my/id/eprint/34441 |
Actions (login required)
 |
View Item |
