Abstract
Hendra virus (HeV) is an emerging zoonotic pathogen harbored by Australian mainland flying foxes. HeV infection can cause lethal disease in humans and horses, and to date all cases of human HeV disease have resulted from contact with infected horses. Currently, diagnosis of acute HeV infections in horses relies on the productive phase of infection when virus shedding may occur. An assay that identifies infected horses during the preclinical phase of infection would reduce the risk of zoonotic viral transmission during management of HeV outbreaks. Having previously shown that the host microRNA (miR)-146a is upregulated in the blood of HeV-infected horses days prior to the detection of viremia, we have profiled miRNAs at the transcriptome-wide level to comprehensively assess differences between infected and uninfected horses. Next-generation sequencing and the miRDeep2 algorithm identified 742 mature miRNA transcripts corresponding to 593 miRNAs in whole blood of six horses (three HeV-infected, three uninfected). Thirty seven miRNAs were differentially expressed in infected horses, two of which were validated by qRT-PCR. This study describes a methodology for the transcriptome-wide profiling of miRNAs in whole blood and supports the notion that measuring host miRNA expression levels may aid infectious disease diagnosis in the future.
Highlights
Hendra virus (HeV) is a negative strand RNA virus belonging to the genus Henipavirus family Paramyxoviridae
Small RNA deep sequencing resulted in 7–11 million raw reads obtained per sample, which have been submitted to the NCBI short read archive (SRA) BioProject PRJNA323953
Host miRNA profiles have been studied in response to acute viral infections associated with prolonged preclinical phases and/or high consequence of infection, such as rabies virus[28] and Ebola virus[29]
Summary
Hendra virus (HeV) is a negative strand RNA virus belonging to the genus Henipavirus family Paramyxoviridae. Current laboratory testing to confirm suspected cases of HeV disease in horses includes virus isolation, detection of viral RNA in clinical or post-mortem specimens or viral antigen detection in tissues taken at necropsy[5]. Serological tests, such as ELISA and virus neutralization test, are available but are of limited use for the diagnosis of acutely infected horses prior to the development of antibodies. We have previously shown that infection of human cells with HeV changes the expression levels of several host-encoded miRNAs, including miR-146a, which was shown to be up-regulated in the blood of three horses experimentally infected with HeV16. Our study identifies a number of miRNAs differentially expressed in the blood of horses infected with HeV, and supports the further development of miRNA profiling to aid henipavirus disease diagnosis
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