title: Diagnostics-in-a-Suitcase: Development of a portable and rapid assay for the detection of the emerging avian influenza A (H7N9) virus.
abstract: BACKGROUND
  In developing countries, equipment necessary for diagnosis is only available in few central laboratories, which are less accessible and of limited capacity to test large numbers of incoming samples. Moreover, the transport conditions of samples are inadequate, therefore leading to unreliable results.
  
  
  OBJECTIVE
  The development of a rapid, inexpensive, and simple test would allow mobile detection of viruses.
  
  
  METHODS
  A suitcase laboratory "Diagnostics-in-a-Suitcase" (56cm×45.5cm×26.5cm) containing all reagents and devices necessary for performing a reverse transcription recombinase polymerase amplification (RT-RPA) assay was developed. As an example, two RT-RPA assays were established for the detection of hemagglutinin (H) and neuraminidase (N) genes of the novel avian influenza (H7N9) virus.
  
  
  RESULTS
  The sensitivities of the H7 and the N9 RT-RPA assays were 10 and 100 RNA molecules, respectively. The assays were performed at a single temperature (42°C). The results were obtained within 2-7min. The H7N9 RT-RPA assays did not show a cross-detection either of any other respiratory viruses affecting humans and/or birds or of the human or chicken genomes. All reagents were used, stored, and transported at ambient temperature, that is, cold chain independent. In addition, the Diagnostics-in-a-Suitcase was operated by a solar-powered battery.
  
  
  CONCLUSIONS
  The developed assay protocol and mobile setup performed well. Moreover, it can be easily implemented to perform diagnoses at airports, quarantine stations, or farms for rapid on-site viral nucleic acid detection.
authors.affiliation: Unit of Infection Models, German Primate Center, Kellnerweg 4, 37077 Goettingen, Germany; Department of Virology, Mansoura University, Algomhoria Street, 35516 Dakahlia, Egypt. Electronic address: abdelwahed@dpz.eu.
authors.forename: Ahmed
authors.initials: A
authors.lastname: Abd El Wahed
authors.affiliation: Institute of Aquaculture, University of Stirling, FK9 4LA Stirling, Scotland, UK.
authors.forename: Manfred
authors.initials: M
authors.lastname: Weidmann
authors.affiliation: Institute of Microbiology and Virology, Brandenburg Medical School Theodor-Fontane, Fehrbelliner Straße 38, 16816 Neuruppin, Brandenburg, Germany.
authors.forename: Frank T
authors.initials: FT
authors.lastname: Hufert
mesh_terms: D000818:Animals; D002645:Chickens; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics; D006801:Humans; D064766:Influenza A Virus, H7N9 Subtype / Q000235:genetics / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis / Q000821:virology; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D009439:Neuraminidase / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011933:Reagent Kits, Diagnostic / Q000191:economics / Q000592:standards; D012680:Sensitivity and Specificity
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011933:Reagent Kits, Diagnostic; D009439:Neuraminidase
keywords: Avian influenza A (H7N9); Diagnostics-in-a-Suitcase; Recombinase polymerase amplification assay