|Appears in Collections:||Aquaculture Journal Articles|
|Peer Review Status:||Refereed|
|Title:||Reverse transcription recombinase polymerase amplification assay for the detection of middle East respiratory syndrome coronavirus|
|Authors:||El, Wahed Ahmed Abd|
Hufert, Frank T
|Citation:||El Wahed AA, Patel P, Heidenreich D, Hufert FT & Weidmann M (2013) Reverse transcription recombinase polymerase amplification assay for the detection of middle East respiratory syndrome coronavirus, PLoS Currents, 5 (Edition 1).|
|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.|
|Rights:||All authors retain copyright of their work. All content in PLOS Currents is open access and available under the terms of the Creative Commons Attribution License. All users of the content are required to cite the original PLOS Currents authors and the source.|
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