Reverse transcription recombinase polymerase amplification assay for the detection of middle East respiratory syndrome coronavirus.
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.
Department of Virology, University Medical Centre, Goettingen, Germany; Department of Virology, Faculty of Veterinary Medicine, Mansoura University, Mansoura, Egypt.
Ahmed
A
Abd El Wahed
Centre for Biological Threats and Special Pathogens, Robert Koch Institute, Berlin, Germany.
Pranav
P
Patel
Department of Virology, University Medical Centre, Goettingen, Germany.
Doris
D
Heidenreich
Department of Virology, University Medical Centre, Goettingen, Germany.
Frank T
FT
Hufert
Department of Virology, University Medical Centre, Goettingen, Germany.
Manfred
M
Weidmann
MERS-coronavirus; RPA; point-of-care assay