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Rapid diagnosis of Hendra virus using a reverse-transcriptase recombinase polymerase amplification and lateral flow detection
Abstract

Rapid diagnosis of Hendra virus using a reverse-transcriptase recombinase polymerase amplification and lateral flow detection

Joanna Kristoffersen, Craig Smith and Joanne Macdonald
XIX International Congress for Tropical Medicine and Malaria Abstract Book, p.1110
International Congress for Tropical Medicine and Malaria (ICTMM), XIX (Brisbane, Australia, 18-Sep-2016–22-Sep-2016)
2016
url
http://tropicalmedicine2016.com/cms/wp-content/uploads/2016/09/ICTMM-2016_Abstract-Book.pdfView
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Abstract

Medical Microbiology
Hendra Virus (HeV) is a highly pathogenic zoonotic disease causing severe illness and mortality in humans and horses. Early detection of this Henipavirus is critical for containing outbreaks, however, confirmatory diagnosis currently requires shipment to a centralized facility that delays diagnosis for several hours or even days. Here we describe the development and testing of a rapid assay for the detection of HeV RNA. A reverse-transcriptase recombinase polymerase assay combined with lateral flow assay (RT-RPA-LFD) was developed that targeted the M gene of the HeV genome. RNA extracted from a wide range of samples from infected and non-infected horses, including nasal, oral, rectal and urogenital swabs, as well as bloods were tested by RT-RPA-LFD and compared to Hendra virus real-time reverse transcriptase PCR where it demonstrated 100% analytical sensitivity and 100% clinical specificity. The HeV RT-RPA LFD was optimized to amplify results in 6 minutes, isothermally at 39 °C, and provides a sensitive, and rapid detection method, amenable for field adaptation. The assay has the potential to dramatically reduce HeV diagnosis times, improve biosecurity surveillance, support a One Health approach, protect attending personnel and reduce prolonged suffering of animals.

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