U.S. Department of Defense


Date of this Version



Journal of Virological Methods 200 (2014) 22–28


This article is a U.S. government work, and is not subject to copyright in the United States.


tHendra and Nipah viruses (HeV and NiV) are closely related zoonotic pathogens of the Paramyxoviridaefamily. Both viruses belong to the Henipavirus genus and cause fatal disease in animals and humans,though only HeV is endemic in Australia. In general and due to the acute nature of the disease, agentdetection by PCR and virus isolation are the primary tools for diagnostic investigations. Assays for thedetection of antibodies against HeV are fit more readily for the purpose of surveillance testing in dis-ease epidemiology and to meet certification requirements in the international movement of horses. Thefirst generation indirect ELISA has been affected by non-specific reactions which must be resolved usingvirus neutralisation serology conducted at laboratory bio-safety level 4 containment (PC4). Recent devel-opments have enabled improvements in the available serology assays. The production of an expressedrecombinant truncated HeV G protein has been utilised in ELISA and in Luminex-based multiplexedmicrosphere assays. In the latter format, two Luminex assays have been developed for use in henipavirusserology: a binding assay (designed for antibody detection and differentiation) and a blocking assay(designed as a surrogate for virus neutralisation). Equine and canine field sera were used to evaluate thetwo Luminex assays relative to ELISA and virus neutralisation serology. Results showed that Luminexassays can be effective as rapid, sensitive and specific tests for the detection of HeV antibody in horse anddog sera. The tests do not require PC4 containment and are appropriate for high throughput applicationsas might be required for disease investigations and other epidemiological surveillance. Also, the resultsshow that the Luminex assays detect effectively HeV vaccine-induced antibodies.