Reverse transcription recombinase polymerase amplification assay for the detection of middle East respiratory syndrome coronavirus
- PMID: 24459611
- PMCID: PMC3871419
- DOI: 10.1371/currents.outbreaks.62df1c7c75ffc96cd59034531e2e8364
Reverse transcription recombinase polymerase amplification assay for the detection of middle East respiratory syndrome coronavirus
Abstract
The emergence of Middle East Respiratory Syndrome Coronavirus (MERS-CoV) in the eastern Mediterranean and imported cases to Europe has alerted public health authorities. Currently, detection of MERS-CoV in patient samples is done by real-time RT-PCR. Samples collected from suspected cases are sent to highly-equipped centralized laboratories for screening. A rapid point-of-care test is needed to allow more widespread mobile detection of the virus directly from patient material. In this study, we describe the development of a reverse transcription isothermal Recombinase Polymerase Amplification (RT-RPA) assay for the identification of MERS-CoV. A partial nucleocapsid gene RNA molecular standard of MERS-coronavirus was used to determine the assay sensitivity. The isothermal (42°C) MERS-CoV RT-RPA was as sensitive as real-time RT-PCR (10 RNA molecules), rapid (3-7 minutes) and mobile (using tubescanner weighing 1kg). The MERS-CoV RT-RPA showed cross-detection neither of any of the RNAs of several coronaviruses and respiratory viruses affecting humans nor of the human genome. The developed isothermal real-time RT-RPA is ideal for rapid mobile molecular MERS-CoV monitoring in acute patients and may also facilitate the search for the animal reservoir of MERS-CoV.
Keywords: MERS-coronavirus; RPA; point-of-care assay.
Figures
Similar articles
-
Rapid and Visual Detection of SARS-CoV-2 RNA Based on Reverse Transcription-Recombinase Polymerase Amplification with Closed Vertical Flow Visualization Strip Assay.Microbiol Spectr. 2023 Feb 14;11(1):e0296622. doi: 10.1128/spectrum.02966-22. Epub 2023 Jan 9. Microbiol Spectr. 2023. PMID: 36622165 Free PMC article.
-
Development of fluorescent reverse transcription loop-mediated isothermal amplification (RT-LAMP) using quenching probes for the detection of the Middle East respiratory syndrome coronavirus.J Virol Methods. 2018 Aug;258:41-48. doi: 10.1016/j.jviromet.2018.05.006. Epub 2018 May 12. J Virol Methods. 2018. PMID: 29763640 Free PMC article.
-
Middle East respiratory syndrome (MERS) coronavirus and dromedaries.Vet J. 2017 Feb;220:75-79. doi: 10.1016/j.tvjl.2016.12.020. Epub 2017 Jan 9. Vet J. 2017. PMID: 28190501 Free PMC article. Review.
-
Animal models of Middle East respiratory syndrome coronavirus infection.Antiviral Res. 2015 Oct;122:28-38. doi: 10.1016/j.antiviral.2015.07.005. Epub 2015 Jul 17. Antiviral Res. 2015. PMID: 26192750 Free PMC article. Review.
-
Detection of Middle East respiratory syndrome coronavirus using reverse transcription loop-mediated isothermal amplification (RT-LAMP).Virol J. 2014 Aug 8;11:139. doi: 10.1186/1743-422X-11-139. Virol J. 2014. PMID: 25103205 Free PMC article.
Cited by
-
A multiplex recombinase polymerase amplification assay combined with CRISPR/Cas12a for the detection of respiratory syncytial virus and respiratory adenovirus.J Int Med Res. 2024 Jan;52(1):3000605231223083. doi: 10.1177/03000605231223083. J Int Med Res. 2024. PMID: 38230675 Free PMC article.
-
Recent progress on rapid diagnosis of COVID-19 by point-of-care testing platforms.Chin Chem Lett. 2023 Jun 15:108688. doi: 10.1016/j.cclet.2023.108688. Online ahead of print. Chin Chem Lett. 2023. PMID: 37362324 Free PMC article. Review.
-
Evaluation of Recombinase Polymerase Amplification assay for monitoring parasite load in patients with kala-azar and post kala-azar dermal leishmaniasis.PLoS Negl Trop Dis. 2023 Apr 19;17(4):e0011231. doi: 10.1371/journal.pntd.0011231. eCollection 2023 Apr. PLoS Negl Trop Dis. 2023. PMID: 37075066 Free PMC article.
-
Rapid detection of human coronavirus NL63 by isothermal reverse transcription recombinase polymerase amplification.J Clin Virol Plus. 2022 Nov;2(4):100115. doi: 10.1016/j.jcvp.2022.100115. Epub 2022 Oct 8. J Clin Virol Plus. 2022. PMID: 36248766 Free PMC article.
-
Quantitative isothermal amplification on paper membranes using amplification nucleation site analysis.Lab Chip. 2022 Jun 14;22(12):2352-2363. doi: 10.1039/d2lc00007e. Lab Chip. 2022. PMID: 35548880 Free PMC article.
References
-
- Zaki AM, van Boheemen S, Bestebroer TM, Osterhaus AD, Fouchier RA. Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia. N Engl J Med. 2012 Nov 8;367(19):1814-20. PubMed PMID:23075143. - PubMed
-
- WHO Middle East Respiratory Syndrome Coronavirus update
-
- Corman VM, Müller MA, Costabel U, Timm J, Binger T, Meyer B, Kreher P, Lattwein E, Eschbach-Bludau M, Nitsche A, Bleicker T, Landt O, Schweiger B, Drexler JF, Osterhaus AD, Haagmans BL, Dittmer U, Bonin F, Wolff T, Drosten C. Assays for laboratory confirmation of novel human coronavirus (hCoV-EMC) infections. Euro Surveill. 2012 Dec 6;17(49). PubMed PMID:23231891. - PubMed
-
- Abd El Wahed A, El-Deeb A, El-Tholoth M, Abd El Kader H, Ahmed A, Hassan S, Hoffmann B, Haas B, Shalaby MA, Hufert FT, Weidmann M. A Portable Reverse Transcription Recombinase Polymerase Amplification Assay for Rapid Detection of Foot-and-Mouth Disease Virus. PLoS One. 2013 Aug 20;8(8):e71642. PubMed PMID:23977101. - PMC - PubMed
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources