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, the necessary equipment for the diagnosis is only available in few central laboratories, which are less accessible and of limited capacity to test large numbers of incoming samples. Moreover, transport conditions of samples are inadequate and therefore lead to unreliable results. Objectives: The development of rapid, inexpensive, and simple test would allow mobile detection of viruses. Study Design: A suitcase laboratory “Diagnostics-in-a-Suitcase” (56 × 45.5 × 26.5 cm) containing all necessary reagents and devices to perform reverse transcription recombinase polymerase amplification (RT-RPA) assay was developed. As an example, Two RT-RPA assays for the detection of hemagglutinin (H) and neuraminidase (N) genes of the novel avian influenza (H7N9) virus were established. Results: Sensitivities were 10 and 100 \{RNA\} molecules for the \{H7\} and the \{N9\} RT-RPA assays, respectively. Assays were performed at a single temperature (42 °C). Results were obtained within 2-7 minutes. The \{H7N9\} RT-RPA assays showed neither a cross-detection of any other respiratory viruses affecting humans and/or birds nor of the human or chicken genomes. All reagents were used, stored, and transported at ambient temperature, i.e. cold chain independent. In addition, the Diagnostics-in-a-Suitcase was operated by a solar power battery. Conclusions: The developed assay protocol and mobile setup performed well. Moreover, it can be easily implemented to perform diagnosis at airport, quarantine stations, or farms for rapid on-site viral nucleic acid detection.